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Protocols of cardiac ion channels INaL Ito1 IKur IKs IK1
Eunjung ParkFDACDERONDDCRP
herg 1
CIPA project-2CIPA project-2
Notes
bull The references are listed in the last slide or if necessary in the bottom of the slide
bull The references in the pulse sequence table are hyperlinked to Pubmed
bull The percentages of channel sources and temperatures in each ion channel were based on selected literatures in this presentation That is not intended to show all relevant published data
Late Sodium Channel Current
bull INaL inhibition Suppress ventricular arrhythmiasndash reduction of intracellular calcium loadingndash decrease of intracellular sodium
bull Potentially selective late INa Blockers at relative low concentrationndash Ranolazine selectivity 9 to 45 fold strong in blocking late vs
peak INa
ndash Amiodarone TTX lidocaine quinidine phenytoin etc
Peak Na(1-2 ms duration)
Phase 2 and early phase 3(100 - 750 ms duration)
Ref Cardiovasc Drugs Ther (2013) 27 79-89
Late sodium channel protocolsRef ID Year Drug Holding (mV) Late Na (ms) Late Na (mV) Peak Na
(ms mV)Frequency
(Hz)
173 1997 -120 100 -20 same 02
100 1998 -150 720 -20 24 -90 to 30
174 1998 R56865 -100 350 -20 No data 02
177 2000 Flecainide -150 Mean (20 and 235) -20 same 5
179 2000 BDF 9198 -140 250 -30 No info
178 2001 Amiodarone -160 200-220 -30 No time info 01
180 2002 -140 750 -40 25 -30
186 2006 -140 2000 na No info 01
205 2009 Ranolazine -140 Mean (46 and 48) -20 24 -20
199 2012 -120 300 -20 No info
160 2013 Ranolazine -130 50 -10 20 -10
195 2013 GS967 -120 220 -20 20 -20
Ex) Ref ID 195peak late
-120 mV complete removal of inactivation
-20 mV
INa enhancer
Pulse20 vs 220 ms
Cardiac potassium channels
ID248 name clone Channel blocker role note
Ito1 Transient outward Kv43Kv14 4-AP (closed state)bullEarly rapid repolarization (Phase I)bullDetermine the height of early plateau
Ito2 is not found in cardiac cells
IKur Ultra rapid delayed rectifier Kv15 4-AP (selective) bullAtrial repolarization
IKs Slow delayed rectifier KCNQ1(KvLQT1) KCNE1 (hminK) Chromanol (293B)
bullAtrial and ventricular repolarizationbullHeart rate dependent
Activation ~0 mV
IKr rapid delayed rectifier Kv111 (hERG) E-4031bullRapid activation and inactivation strong inward rectificationbullCardiac APD and refractoriness
Activation ~-30mV[K+] dependent
IK1 Inward rectifier Kir21 Ba2+ (IC50= 20 uM)
bullMaintain the resting membrane potentialbullControl excitabilitybullInitiate the final repolarization phase
Nav15
Kv43 (Ito f) Kv15 (IKur)
KvLQT1minK (IKs)
Cav12 (ICaL) hERG (IKr)
Cav32 (IcaT)
Kir 21 (IK1)
Summary of Ito1 protocolsbull Distribution Atrial PF epicardial M cells gt endocardialbull Pharmacology A common finding in cardiac disease is reduction in Ito1 density with a loss
of heterogeneity of Ito expression and associated action potential prolongation
bull Channel sources1 Transfected cell lines CHO L cell COS-7 cell HEK 293 and Xenopus Oocyte (818 = 44)2 Myocyte (56) human AM (3) rat VM (4) Canine VM 92) and rabbit VM (1)
Guinea Pig (0 very low expression of Ito)
bull Temperature ndash RT = 1215 (80)ndash near or real physiological 315 (20)
1 Pulse sequencesndash Holding -90 to -40 (mode -80 mV) ndash Activation 0 to 75 mV (mode 50 mV)ndash Duration of activation 150 ndash 500 ms (mode 300 ms)
bull Ito measurement peak current or after 70ms of activation
Pulse sequences Ito
Ito (Kir 43)
ID ChannelCurrent
Speciescells CPD Temp Vh(mV)
Prepulse(ms mV)
Activation(ms mV)
Voltage -Dependence(ms mV)
Intervalfrequency
47 hKv43hKChIP22 CHO Methadone RT -90 300 40
49 rKv43 MouseL cell Vanoxerine RT -80 300 20 15s
50 hKv43 HEK293 Alfuzosin RT -80 300 20 15s
206 Ito RatVM Zacopride 24 -40 na 500 -40 to 80 (10)
210 Ito HumanAM Cyamemazine
32-34 -50 500 60
214 Kv43 MonkeyCOS7 Pentamidine na -80 na 500 -40 to 60
215 Ito CanineVM Pioglitazone na -80 5ms -40 na 50
220 Ito RatVM Ajmaline RT -75 300 60
221 rKv42 rKv43 HEK293 RSD1235 25 -80 Brief -50 250 60 1Hz
222 Ito RatVM FK-506 22-24 -80 na 50 1000 -50 to 70 01Hz
224 hKv43 HEK293 Allitridi 22-23 -80 300 60 300 -80 to 60
245 Ito HumanAM DPO RT -80 150 40
256 Ito RabbitVM Carvedilol 34 -60 300 40 300 -50 to 60
263 Kv43 CHO (transient) Spironolactone canrenoic acid
RT -80 250 50 250 -90 to 50 10s
267 Ito ratVM Ebastine na -90 na 75 na -30 to 75
273 Ito CanineVM Thymol 37 -80 5ms- 40 400 50 400 -10 to 60
281 rKv14 Xenopus Oocyte Haloperidol 22 -80 300 0
283 Ito HumanAM Clotrimaxole 21-22 -50 300 50 300 -40 to 60 02Hz
VM= ventricular myocyte AM= Atrial myocyte
Kinetics Pulse Sequences Ito
ID channel cells kinetics Vh (mV)
1st (ms mv)
2nd (ms mV)
3rd (ms mv)
4th (ms mV)
224 hKv43 HEK293 Inactivation -80 -10 to 60
Channel availability -80 na -100 to 40 300 50
Activation conductance -80 8 na na
Recovery time from inactivation -80 30s 50 t -80 300 50 -80
Close state inactivation -90 300 50 3000 -90 t -50 300 50
283 Ito HumanAM Voltage dependence of activation and inactivation
-80 250 -100 to 30 (10) na 50
Recovery time from inactivation -80 30 -40 300 50 t -80 300 50
Extracellular buffer solution of ItoID NaCl NMDG KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-
acetate Blocker pH pH buffer
47 49 50 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 20 033 CdCl2 (01 CaL) BaCl2 (02 Ik1) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 CdCl2
214 140 4 2 1 10 74 NaOH
215 144 56 25 12 5 11 74 na
220 135 54 09 09 10 10 033 CoCl2 (2) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
222 135 4 18 1 2 11 TTX (001)Co++ (2) 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245 132 4 12 10 111 Nisoldipine (04 uM) na na
256 143 54 18 05 5 56 025 Nisoldipine (0003) or CoCl2 (2) and TTX (001) 74 NaOH
263-1 130 4 1 1 10 10 74 NaOH
263-2 140 4 1 1 10 10 TTX (003) CoCl2 (2) 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 140 5 18 1 6 6 Nifedipine (0005) 74 NaOH
283 140 5 1 5 10 033 BaCl2 (01) CdCl2 (02) 74 NaOH
NMDG N-methyl-D-glucamine 263-1 CHO cells 263-2 mouse VM
Pipette solution of ItoID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phospho creatinne
NaCl CaCl2 KOH KH2PO4 pH pH buffer
47 49 50
130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
214 110 142 10 4 10 517 72 KOH
215 45 100 1 10 na 5 72
220 25 130 1 1 5 01 5 3 725 KOH
221 130 1 5 4 01 10 72 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245 20 110 5 5 10 72
256 80 40 10 5 5 5 065 60 72
263 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
Summary of IKur (Kv1531) protocolsbull Distribution a major repolarizing current in human atria but seems to play no
role in the ventricle bull Pharmacology selective IKur blockers to restore and maintain sinus rhythm
without a risk of ventricular pro-arrhythmia
bull Channel sources1 Transfected cell lines CHO L cells HEK 293 and Xenopus Oocyte (710 =
70)2 Myocyte (30) species ndash human AM and mouse VM
bull Temperature RT = 100bull Pulse sequences
ndash Holding range -80 to -50 mV (mode -80 mV)ndash Activation 20 - 60 mV (mode 50 mV)ndash Duration of activation 150 ndash 1000 ms (mode 300 ms)
1 IKur measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKurID Channel
CurrentSpeciescells CPD Temp Vh
(mV)Prepulse Activation
(ms mV)Tail current
Voltage -Dependence(ms mV)
Interval (s)
49 hKv15 MouseL cell vanoxerine RT -80 300 20 10s
221 Kv15 HEK293 RSD1235 25 -80 200 60
224 hKv15 HEK293 Allitridi 22-23 -80 300 50 300 -40 to 60
245-1 hKv15 CHO DPO RT -80 150 40 na -30
245-2 Ikur HumanAM DPO RT -50 300 40 na -30
245-3 hKv15 XO DPO na -80 na 40
262 hKv15 CHO SSR149744C RT -70 1000 30 20s
263 hKv15 Mouse LtK- Spironolactonecanrenoic acid
RT -80 500 60 500 -40 500 -80 to 60
263 Ikur MouseVM Spironolactonecanrenoic acid
RT -80 200 40 500 50 10s
283 Ikur HumanAM clotrimazole 21-22 -50 100 40 150 50 na -30 150 -40 to 50
Ikur (Kv15)
Extracellular buffer solution of IKurID NaCl NMCG K-
gluconate KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-acetate Blocker pH pH
buffer
49 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 10 033 CdCl2 (01) BaCl2 (02) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245-1 132 4 18 12 10 111 72 na
245-2 132 4 12 10 111 nisoldipine (04)
245-3 116 4 10 725
262 140 54 2 2 15 10 74 NaOH
263 130 4 1 1 10 10 74 NaOH
281 96 2 18 1 5 74 NaOH
283 140 5 1 1 5 10 033BaCl2 (02 Ik1) CdCl2 (Ica)
74 NaOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Pipette solution of IKurID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phosphocreatinne
NaCl CaCl2 K-gluconate KH2PO4 pH pH
buffer
49 130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
221 130 1 5 4 01 10 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245-1 15 32 5 5 5 119 735
245-2 20 110 5 5 10 72
245-3 4 10 003 116 725
262 140 2 11 10 2 73 KOH
263 42 80 5 5 5 3 10 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Summary of Iks protocolsbull Distribution all cell types but less in midmyocardial myocytes
bull Pharmacology Atrial and ventricular repolarizationHeart rate dependentbull subunit KCNQ1 (channel forming) Kv71 KvLQT1bull subunit KCNE1 (accessary unit activation of KCNQ1) minK (minimum potassium
subunit)
bull Channel sources1 transfected cell lines CHO HEK 293 Cos-7 cells and Xenopus Oocyte (2136 = 58)2 Myocyte (42) species ndash Canine VM Guinea pig VM rabbit VM and mouse VM (mode
GP)
bull Temperature RT = 1629 55 physiological temperature = 45
bull Pulse sequencesndash Holding potential range = -90 to -30 (mode -80)ndash Depolarization potential 20-60 mV ndash Duration of depolarization 50 ms ndash 10s (mode 2s)ndash Repolarization potential -80 to -10 mV (mode -40 mV)ndash Duration of repolarization 05 ndash 5s (mode 2s)
bull IKs measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation (s mV)
deactivation(s mV)
Interval (s)
47 hKvLQT1hminK CHO RT -80 2 40 01 Hz
49 hKv711hKCNE1 CHO RT -80 2 20 05 -40 15s
50 hKv711hKCNE1 HEk293 RT -80 2 20 05 -40 15s
184 IKs canineepiardial cells 37 -50 2 40 na -30 na
214 Kv711minK HEK293 na -80 5 -60 to 60 5s -50 na
215 IKs caninemyocyte na -40 5 50
224 hKCNQ1hKCNE1 HEK293 22-23 -80 3 -60 to 60 (20) na -50 na
229 hCNE1-hKCNQ1 Transgenic mousemyocyte RT -75 20 -40(Na block) 2 60 2 -40 006 Hz
230 KCNQ1KCNE1 XenopusOocyte 20-22 -80 5 30 na -70
232 KCNQ1KCNE1 HEK293 36 -80 2 -80 to 110 (10) 2 -60 6s
233 KCNQ1KCNE1 CHO na -80 80 -90 3 20 1 -50 005Hz
237 KCNQ1 CHO na -80 na -80 to 60 1 -40
239 KCNQ1KCNE1 HEK293 RT -70 1 -70 to 60 na -40
241 KCNQ1 andor minK MouseCOS-7 22-25 -80 3 -120 to 60 (20) 2 -30
242 hKvLQT1hminK HEK293 19-36 -80 3 -50 to 60 (10) 5 -50
242 hKvLQT1hminK HEK293 19-36 -50 1 or 2 50 na -50 10s
244 KCNQ1 andor KCNE1 CHO na -90 3 -70 to 70 (10) na -60
245 IKs GPMyocyte 35 -50 1 50
249 KCNQ1KCNE1 HEK293 33-34 -40 1 -40 to 80 (10) na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation(s mV)
Voltage dependent(s mV)
Deactivation(s mV)
Interval (s)
251 IKs GPMyocyte 22-2535-37 -40 2 60 2 -40 to 80 (20) na -40
252 IKs GPMyocyte 35 -80 200 -50 3 60 2 -40 60s
252 hminK XenopusOocyte 35 -80 10 20 10 -40 to 60 2 -80
256 IKs Rabbitmyocyte 34 -50 3 -40 to 50 na
260 hKCNQ1hKCNE1 HEK293 RT -80 25 -70 to 100 (10) 25 -40 15s
262 IKs GPMyocyte 35 -80 1 40 5 -10
263 Kv71 and minK CHO RT -80 2 40 na -40
264 IKs GPmyocyte na -80 200 -40 05 50 na -40
267 IKs GPmyocyte na -90 na 0 na 40 01- 015 0-80 -90
273 IKs Caninemyocyte 37 -40 3 50 na -40
274 IKs GPmyocyte 36 -30 05 50 or 2 70 05 (2) -30 to 70 na -30
275 KCNQ1KCNE1 CHO 21 -80 1 40 01Hz
276 hKCNQ1hminK CHO 22 -80 6 40 na -80 005Hz
278 IKs Canine myocyte 35-37 -50 3 -50 to 50 2s -40
283 KCNQ1KCNE1 HEK293 21-22 -80 3 40 3 -80 to 40 na -40
286 IKs GPmyocyte 34-36 -50 005 30 005 ndash3 -50 to 30 na -50
287 Iks GPmyocyte 36 -80 na -40 2 60 2 -40 to 80 na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
47 49 50
137 4 18 1 10 10 74 NaOH
184 145 4 18 18 10 11 NaH2PO4 1 E4031 (5uM) Cadmium (300 uM) 74 NaOH
214 140 4 2 1 10 74 NaOH
215 144 5 56 25 5 Dextrose 11 Nifedipine (5 uM) E4031 (1 uM) 74 NaOH
224 140 54 18 1 10 10 NaH2PO4 033 73 NaOH
229 132 48 2 12 10 5 E4031 nisoldipine 74 NaOH
230 96 2 18 1 5 74 NaOH
232 140 54 18 1 5 55 74 NaOH
233 17 120 4 2 1 10 10 74 NaOH
237 130 5 18 1 10 Na-Acetate 28 74 NaOH
239 150 54 18 4 10 74 NaOH
241 135 5 25 12 5 10 74 NaOH
242 132 4 18 12 10 111 74 NaOH
244 140 4 18 12 55 11 74 NaOH
249 140 54 18 05 5 55 NaH2PO4 033 74 NaOH
250 140 54 18 1 5 10 74 NaOH
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
251 1 1 2 10 N-methly-d-glucamine 132
lanthanum chloride 005 mMcadmium chloride 02 mM 74 HCl
252 140 47 13 1 10 10 74 NaOH
256 09 5 N-methly-d-glucamine 149
Nisoldipine 3 uME4031 10 uM 74 HCl
260 140 54 18 1 5 10 NaH2PO4 03 73 NaOH
262 137 4 18 2 10 115 Pyrubic acid 5 Thiamine 01 74 NaOH
263 130 4 1 1 10 10 TTX 003 mM CoCl2 2 mM 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10 KH2PO4 12 MgSO4 12 NaHCO3 44
Nifetidine 5 uM 4-aminopyridine 3 mM 72 NaOH
274 140 1 5 10 Cd2+ 02 mM 74 NaOH
275 130 54 18 1 10 5 74 NaOH
278 136 54 1 1 5 Dextrose 10 NaH2PO4 033 E4031 5 uM 74 NaOH
283 140 5 1 1 5 10 NaH2PO4 033 74 NaOH
286 09 5 5 N-methly-d-glucamine 149
Nisoldipine 04 uME4031 5 uM 74 NaOH
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
49 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
214 110 142 10 4 4 517 72 KOH
215 45 100 1 10 na 5 72
224 20 110 1 5 5 01 10 5 72 KOH
229 110 11 5 10 55 73
230 3000
232 20 125 10 10
233 140 1 1 20 73 KOH
237 10 125 1 10 5 10 10 72 KOH
239 30 130 01 4 10 74
241 117 2 11 11 10 1 72
242 15 119 32 5 5 5 74 KOH
244 130 1 5 5 10 74 KOH
249 130 5 5 01 5 5 74 HCl
250 80 40 10 5 01 5 40 065 72 HCl
251 50 60 1 11 5 10 1 74 KOH
252 140 11 1 10 10 72 KOH
amphotericin-B
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
256 80 40 10 5 5 5 065 72
262 150 1 5 4 5 5 73 KOH
263^ 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
274 40 106 1 5 4 5 72 KOH
276 20 110 1 10 5 10 10 1 72 KOH
278 20 110 1 10 5 01 10 5 72 KOH
283 20 110 1 5 5 01 1 5 72 KOH
286 50 1 5 5 01 5 2 10 72 KOH
^phosphatidylinositol 45-diphosphate (5 mgml 1) was added to the internal solution to avoid the rundown of the current 70 L-aspartic acid 70 KOH
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Notes
bull The references are listed in the last slide or if necessary in the bottom of the slide
bull The references in the pulse sequence table are hyperlinked to Pubmed
bull The percentages of channel sources and temperatures in each ion channel were based on selected literatures in this presentation That is not intended to show all relevant published data
Late Sodium Channel Current
bull INaL inhibition Suppress ventricular arrhythmiasndash reduction of intracellular calcium loadingndash decrease of intracellular sodium
bull Potentially selective late INa Blockers at relative low concentrationndash Ranolazine selectivity 9 to 45 fold strong in blocking late vs
peak INa
ndash Amiodarone TTX lidocaine quinidine phenytoin etc
Peak Na(1-2 ms duration)
Phase 2 and early phase 3(100 - 750 ms duration)
Ref Cardiovasc Drugs Ther (2013) 27 79-89
Late sodium channel protocolsRef ID Year Drug Holding (mV) Late Na (ms) Late Na (mV) Peak Na
(ms mV)Frequency
(Hz)
173 1997 -120 100 -20 same 02
100 1998 -150 720 -20 24 -90 to 30
174 1998 R56865 -100 350 -20 No data 02
177 2000 Flecainide -150 Mean (20 and 235) -20 same 5
179 2000 BDF 9198 -140 250 -30 No info
178 2001 Amiodarone -160 200-220 -30 No time info 01
180 2002 -140 750 -40 25 -30
186 2006 -140 2000 na No info 01
205 2009 Ranolazine -140 Mean (46 and 48) -20 24 -20
199 2012 -120 300 -20 No info
160 2013 Ranolazine -130 50 -10 20 -10
195 2013 GS967 -120 220 -20 20 -20
Ex) Ref ID 195peak late
-120 mV complete removal of inactivation
-20 mV
INa enhancer
Pulse20 vs 220 ms
Cardiac potassium channels
ID248 name clone Channel blocker role note
Ito1 Transient outward Kv43Kv14 4-AP (closed state)bullEarly rapid repolarization (Phase I)bullDetermine the height of early plateau
Ito2 is not found in cardiac cells
IKur Ultra rapid delayed rectifier Kv15 4-AP (selective) bullAtrial repolarization
IKs Slow delayed rectifier KCNQ1(KvLQT1) KCNE1 (hminK) Chromanol (293B)
bullAtrial and ventricular repolarizationbullHeart rate dependent
Activation ~0 mV
IKr rapid delayed rectifier Kv111 (hERG) E-4031bullRapid activation and inactivation strong inward rectificationbullCardiac APD and refractoriness
Activation ~-30mV[K+] dependent
IK1 Inward rectifier Kir21 Ba2+ (IC50= 20 uM)
bullMaintain the resting membrane potentialbullControl excitabilitybullInitiate the final repolarization phase
Nav15
Kv43 (Ito f) Kv15 (IKur)
KvLQT1minK (IKs)
Cav12 (ICaL) hERG (IKr)
Cav32 (IcaT)
Kir 21 (IK1)
Summary of Ito1 protocolsbull Distribution Atrial PF epicardial M cells gt endocardialbull Pharmacology A common finding in cardiac disease is reduction in Ito1 density with a loss
of heterogeneity of Ito expression and associated action potential prolongation
bull Channel sources1 Transfected cell lines CHO L cell COS-7 cell HEK 293 and Xenopus Oocyte (818 = 44)2 Myocyte (56) human AM (3) rat VM (4) Canine VM 92) and rabbit VM (1)
Guinea Pig (0 very low expression of Ito)
bull Temperature ndash RT = 1215 (80)ndash near or real physiological 315 (20)
1 Pulse sequencesndash Holding -90 to -40 (mode -80 mV) ndash Activation 0 to 75 mV (mode 50 mV)ndash Duration of activation 150 ndash 500 ms (mode 300 ms)
bull Ito measurement peak current or after 70ms of activation
Pulse sequences Ito
Ito (Kir 43)
ID ChannelCurrent
Speciescells CPD Temp Vh(mV)
Prepulse(ms mV)
Activation(ms mV)
Voltage -Dependence(ms mV)
Intervalfrequency
47 hKv43hKChIP22 CHO Methadone RT -90 300 40
49 rKv43 MouseL cell Vanoxerine RT -80 300 20 15s
50 hKv43 HEK293 Alfuzosin RT -80 300 20 15s
206 Ito RatVM Zacopride 24 -40 na 500 -40 to 80 (10)
210 Ito HumanAM Cyamemazine
32-34 -50 500 60
214 Kv43 MonkeyCOS7 Pentamidine na -80 na 500 -40 to 60
215 Ito CanineVM Pioglitazone na -80 5ms -40 na 50
220 Ito RatVM Ajmaline RT -75 300 60
221 rKv42 rKv43 HEK293 RSD1235 25 -80 Brief -50 250 60 1Hz
222 Ito RatVM FK-506 22-24 -80 na 50 1000 -50 to 70 01Hz
224 hKv43 HEK293 Allitridi 22-23 -80 300 60 300 -80 to 60
245 Ito HumanAM DPO RT -80 150 40
256 Ito RabbitVM Carvedilol 34 -60 300 40 300 -50 to 60
263 Kv43 CHO (transient) Spironolactone canrenoic acid
RT -80 250 50 250 -90 to 50 10s
267 Ito ratVM Ebastine na -90 na 75 na -30 to 75
273 Ito CanineVM Thymol 37 -80 5ms- 40 400 50 400 -10 to 60
281 rKv14 Xenopus Oocyte Haloperidol 22 -80 300 0
283 Ito HumanAM Clotrimaxole 21-22 -50 300 50 300 -40 to 60 02Hz
VM= ventricular myocyte AM= Atrial myocyte
Kinetics Pulse Sequences Ito
ID channel cells kinetics Vh (mV)
1st (ms mv)
2nd (ms mV)
3rd (ms mv)
4th (ms mV)
224 hKv43 HEK293 Inactivation -80 -10 to 60
Channel availability -80 na -100 to 40 300 50
Activation conductance -80 8 na na
Recovery time from inactivation -80 30s 50 t -80 300 50 -80
Close state inactivation -90 300 50 3000 -90 t -50 300 50
283 Ito HumanAM Voltage dependence of activation and inactivation
-80 250 -100 to 30 (10) na 50
Recovery time from inactivation -80 30 -40 300 50 t -80 300 50
Extracellular buffer solution of ItoID NaCl NMDG KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-
acetate Blocker pH pH buffer
47 49 50 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 20 033 CdCl2 (01 CaL) BaCl2 (02 Ik1) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 CdCl2
214 140 4 2 1 10 74 NaOH
215 144 56 25 12 5 11 74 na
220 135 54 09 09 10 10 033 CoCl2 (2) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
222 135 4 18 1 2 11 TTX (001)Co++ (2) 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245 132 4 12 10 111 Nisoldipine (04 uM) na na
256 143 54 18 05 5 56 025 Nisoldipine (0003) or CoCl2 (2) and TTX (001) 74 NaOH
263-1 130 4 1 1 10 10 74 NaOH
263-2 140 4 1 1 10 10 TTX (003) CoCl2 (2) 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 140 5 18 1 6 6 Nifedipine (0005) 74 NaOH
283 140 5 1 5 10 033 BaCl2 (01) CdCl2 (02) 74 NaOH
NMDG N-methyl-D-glucamine 263-1 CHO cells 263-2 mouse VM
Pipette solution of ItoID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phospho creatinne
NaCl CaCl2 KOH KH2PO4 pH pH buffer
47 49 50
130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
214 110 142 10 4 10 517 72 KOH
215 45 100 1 10 na 5 72
220 25 130 1 1 5 01 5 3 725 KOH
221 130 1 5 4 01 10 72 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245 20 110 5 5 10 72
256 80 40 10 5 5 5 065 60 72
263 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
Summary of IKur (Kv1531) protocolsbull Distribution a major repolarizing current in human atria but seems to play no
role in the ventricle bull Pharmacology selective IKur blockers to restore and maintain sinus rhythm
without a risk of ventricular pro-arrhythmia
bull Channel sources1 Transfected cell lines CHO L cells HEK 293 and Xenopus Oocyte (710 =
70)2 Myocyte (30) species ndash human AM and mouse VM
bull Temperature RT = 100bull Pulse sequences
ndash Holding range -80 to -50 mV (mode -80 mV)ndash Activation 20 - 60 mV (mode 50 mV)ndash Duration of activation 150 ndash 1000 ms (mode 300 ms)
1 IKur measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKurID Channel
CurrentSpeciescells CPD Temp Vh
(mV)Prepulse Activation
(ms mV)Tail current
Voltage -Dependence(ms mV)
Interval (s)
49 hKv15 MouseL cell vanoxerine RT -80 300 20 10s
221 Kv15 HEK293 RSD1235 25 -80 200 60
224 hKv15 HEK293 Allitridi 22-23 -80 300 50 300 -40 to 60
245-1 hKv15 CHO DPO RT -80 150 40 na -30
245-2 Ikur HumanAM DPO RT -50 300 40 na -30
245-3 hKv15 XO DPO na -80 na 40
262 hKv15 CHO SSR149744C RT -70 1000 30 20s
263 hKv15 Mouse LtK- Spironolactonecanrenoic acid
RT -80 500 60 500 -40 500 -80 to 60
263 Ikur MouseVM Spironolactonecanrenoic acid
RT -80 200 40 500 50 10s
283 Ikur HumanAM clotrimazole 21-22 -50 100 40 150 50 na -30 150 -40 to 50
Ikur (Kv15)
Extracellular buffer solution of IKurID NaCl NMCG K-
gluconate KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-acetate Blocker pH pH
buffer
49 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 10 033 CdCl2 (01) BaCl2 (02) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245-1 132 4 18 12 10 111 72 na
245-2 132 4 12 10 111 nisoldipine (04)
245-3 116 4 10 725
262 140 54 2 2 15 10 74 NaOH
263 130 4 1 1 10 10 74 NaOH
281 96 2 18 1 5 74 NaOH
283 140 5 1 1 5 10 033BaCl2 (02 Ik1) CdCl2 (Ica)
74 NaOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Pipette solution of IKurID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phosphocreatinne
NaCl CaCl2 K-gluconate KH2PO4 pH pH
buffer
49 130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
221 130 1 5 4 01 10 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245-1 15 32 5 5 5 119 735
245-2 20 110 5 5 10 72
245-3 4 10 003 116 725
262 140 2 11 10 2 73 KOH
263 42 80 5 5 5 3 10 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Summary of Iks protocolsbull Distribution all cell types but less in midmyocardial myocytes
bull Pharmacology Atrial and ventricular repolarizationHeart rate dependentbull subunit KCNQ1 (channel forming) Kv71 KvLQT1bull subunit KCNE1 (accessary unit activation of KCNQ1) minK (minimum potassium
subunit)
bull Channel sources1 transfected cell lines CHO HEK 293 Cos-7 cells and Xenopus Oocyte (2136 = 58)2 Myocyte (42) species ndash Canine VM Guinea pig VM rabbit VM and mouse VM (mode
GP)
bull Temperature RT = 1629 55 physiological temperature = 45
bull Pulse sequencesndash Holding potential range = -90 to -30 (mode -80)ndash Depolarization potential 20-60 mV ndash Duration of depolarization 50 ms ndash 10s (mode 2s)ndash Repolarization potential -80 to -10 mV (mode -40 mV)ndash Duration of repolarization 05 ndash 5s (mode 2s)
bull IKs measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation (s mV)
deactivation(s mV)
Interval (s)
47 hKvLQT1hminK CHO RT -80 2 40 01 Hz
49 hKv711hKCNE1 CHO RT -80 2 20 05 -40 15s
50 hKv711hKCNE1 HEk293 RT -80 2 20 05 -40 15s
184 IKs canineepiardial cells 37 -50 2 40 na -30 na
214 Kv711minK HEK293 na -80 5 -60 to 60 5s -50 na
215 IKs caninemyocyte na -40 5 50
224 hKCNQ1hKCNE1 HEK293 22-23 -80 3 -60 to 60 (20) na -50 na
229 hCNE1-hKCNQ1 Transgenic mousemyocyte RT -75 20 -40(Na block) 2 60 2 -40 006 Hz
230 KCNQ1KCNE1 XenopusOocyte 20-22 -80 5 30 na -70
232 KCNQ1KCNE1 HEK293 36 -80 2 -80 to 110 (10) 2 -60 6s
233 KCNQ1KCNE1 CHO na -80 80 -90 3 20 1 -50 005Hz
237 KCNQ1 CHO na -80 na -80 to 60 1 -40
239 KCNQ1KCNE1 HEK293 RT -70 1 -70 to 60 na -40
241 KCNQ1 andor minK MouseCOS-7 22-25 -80 3 -120 to 60 (20) 2 -30
242 hKvLQT1hminK HEK293 19-36 -80 3 -50 to 60 (10) 5 -50
242 hKvLQT1hminK HEK293 19-36 -50 1 or 2 50 na -50 10s
244 KCNQ1 andor KCNE1 CHO na -90 3 -70 to 70 (10) na -60
245 IKs GPMyocyte 35 -50 1 50
249 KCNQ1KCNE1 HEK293 33-34 -40 1 -40 to 80 (10) na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation(s mV)
Voltage dependent(s mV)
Deactivation(s mV)
Interval (s)
251 IKs GPMyocyte 22-2535-37 -40 2 60 2 -40 to 80 (20) na -40
252 IKs GPMyocyte 35 -80 200 -50 3 60 2 -40 60s
252 hminK XenopusOocyte 35 -80 10 20 10 -40 to 60 2 -80
256 IKs Rabbitmyocyte 34 -50 3 -40 to 50 na
260 hKCNQ1hKCNE1 HEK293 RT -80 25 -70 to 100 (10) 25 -40 15s
262 IKs GPMyocyte 35 -80 1 40 5 -10
263 Kv71 and minK CHO RT -80 2 40 na -40
264 IKs GPmyocyte na -80 200 -40 05 50 na -40
267 IKs GPmyocyte na -90 na 0 na 40 01- 015 0-80 -90
273 IKs Caninemyocyte 37 -40 3 50 na -40
274 IKs GPmyocyte 36 -30 05 50 or 2 70 05 (2) -30 to 70 na -30
275 KCNQ1KCNE1 CHO 21 -80 1 40 01Hz
276 hKCNQ1hminK CHO 22 -80 6 40 na -80 005Hz
278 IKs Canine myocyte 35-37 -50 3 -50 to 50 2s -40
283 KCNQ1KCNE1 HEK293 21-22 -80 3 40 3 -80 to 40 na -40
286 IKs GPmyocyte 34-36 -50 005 30 005 ndash3 -50 to 30 na -50
287 Iks GPmyocyte 36 -80 na -40 2 60 2 -40 to 80 na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
47 49 50
137 4 18 1 10 10 74 NaOH
184 145 4 18 18 10 11 NaH2PO4 1 E4031 (5uM) Cadmium (300 uM) 74 NaOH
214 140 4 2 1 10 74 NaOH
215 144 5 56 25 5 Dextrose 11 Nifedipine (5 uM) E4031 (1 uM) 74 NaOH
224 140 54 18 1 10 10 NaH2PO4 033 73 NaOH
229 132 48 2 12 10 5 E4031 nisoldipine 74 NaOH
230 96 2 18 1 5 74 NaOH
232 140 54 18 1 5 55 74 NaOH
233 17 120 4 2 1 10 10 74 NaOH
237 130 5 18 1 10 Na-Acetate 28 74 NaOH
239 150 54 18 4 10 74 NaOH
241 135 5 25 12 5 10 74 NaOH
242 132 4 18 12 10 111 74 NaOH
244 140 4 18 12 55 11 74 NaOH
249 140 54 18 05 5 55 NaH2PO4 033 74 NaOH
250 140 54 18 1 5 10 74 NaOH
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
251 1 1 2 10 N-methly-d-glucamine 132
lanthanum chloride 005 mMcadmium chloride 02 mM 74 HCl
252 140 47 13 1 10 10 74 NaOH
256 09 5 N-methly-d-glucamine 149
Nisoldipine 3 uME4031 10 uM 74 HCl
260 140 54 18 1 5 10 NaH2PO4 03 73 NaOH
262 137 4 18 2 10 115 Pyrubic acid 5 Thiamine 01 74 NaOH
263 130 4 1 1 10 10 TTX 003 mM CoCl2 2 mM 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10 KH2PO4 12 MgSO4 12 NaHCO3 44
Nifetidine 5 uM 4-aminopyridine 3 mM 72 NaOH
274 140 1 5 10 Cd2+ 02 mM 74 NaOH
275 130 54 18 1 10 5 74 NaOH
278 136 54 1 1 5 Dextrose 10 NaH2PO4 033 E4031 5 uM 74 NaOH
283 140 5 1 1 5 10 NaH2PO4 033 74 NaOH
286 09 5 5 N-methly-d-glucamine 149
Nisoldipine 04 uME4031 5 uM 74 NaOH
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
49 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
214 110 142 10 4 4 517 72 KOH
215 45 100 1 10 na 5 72
224 20 110 1 5 5 01 10 5 72 KOH
229 110 11 5 10 55 73
230 3000
232 20 125 10 10
233 140 1 1 20 73 KOH
237 10 125 1 10 5 10 10 72 KOH
239 30 130 01 4 10 74
241 117 2 11 11 10 1 72
242 15 119 32 5 5 5 74 KOH
244 130 1 5 5 10 74 KOH
249 130 5 5 01 5 5 74 HCl
250 80 40 10 5 01 5 40 065 72 HCl
251 50 60 1 11 5 10 1 74 KOH
252 140 11 1 10 10 72 KOH
amphotericin-B
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
256 80 40 10 5 5 5 065 72
262 150 1 5 4 5 5 73 KOH
263^ 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
274 40 106 1 5 4 5 72 KOH
276 20 110 1 10 5 10 10 1 72 KOH
278 20 110 1 10 5 01 10 5 72 KOH
283 20 110 1 5 5 01 1 5 72 KOH
286 50 1 5 5 01 5 2 10 72 KOH
^phosphatidylinositol 45-diphosphate (5 mgml 1) was added to the internal solution to avoid the rundown of the current 70 L-aspartic acid 70 KOH
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Late Sodium Channel Current
bull INaL inhibition Suppress ventricular arrhythmiasndash reduction of intracellular calcium loadingndash decrease of intracellular sodium
bull Potentially selective late INa Blockers at relative low concentrationndash Ranolazine selectivity 9 to 45 fold strong in blocking late vs
peak INa
ndash Amiodarone TTX lidocaine quinidine phenytoin etc
Peak Na(1-2 ms duration)
Phase 2 and early phase 3(100 - 750 ms duration)
Ref Cardiovasc Drugs Ther (2013) 27 79-89
Late sodium channel protocolsRef ID Year Drug Holding (mV) Late Na (ms) Late Na (mV) Peak Na
(ms mV)Frequency
(Hz)
173 1997 -120 100 -20 same 02
100 1998 -150 720 -20 24 -90 to 30
174 1998 R56865 -100 350 -20 No data 02
177 2000 Flecainide -150 Mean (20 and 235) -20 same 5
179 2000 BDF 9198 -140 250 -30 No info
178 2001 Amiodarone -160 200-220 -30 No time info 01
180 2002 -140 750 -40 25 -30
186 2006 -140 2000 na No info 01
205 2009 Ranolazine -140 Mean (46 and 48) -20 24 -20
199 2012 -120 300 -20 No info
160 2013 Ranolazine -130 50 -10 20 -10
195 2013 GS967 -120 220 -20 20 -20
Ex) Ref ID 195peak late
-120 mV complete removal of inactivation
-20 mV
INa enhancer
Pulse20 vs 220 ms
Cardiac potassium channels
ID248 name clone Channel blocker role note
Ito1 Transient outward Kv43Kv14 4-AP (closed state)bullEarly rapid repolarization (Phase I)bullDetermine the height of early plateau
Ito2 is not found in cardiac cells
IKur Ultra rapid delayed rectifier Kv15 4-AP (selective) bullAtrial repolarization
IKs Slow delayed rectifier KCNQ1(KvLQT1) KCNE1 (hminK) Chromanol (293B)
bullAtrial and ventricular repolarizationbullHeart rate dependent
Activation ~0 mV
IKr rapid delayed rectifier Kv111 (hERG) E-4031bullRapid activation and inactivation strong inward rectificationbullCardiac APD and refractoriness
Activation ~-30mV[K+] dependent
IK1 Inward rectifier Kir21 Ba2+ (IC50= 20 uM)
bullMaintain the resting membrane potentialbullControl excitabilitybullInitiate the final repolarization phase
Nav15
Kv43 (Ito f) Kv15 (IKur)
KvLQT1minK (IKs)
Cav12 (ICaL) hERG (IKr)
Cav32 (IcaT)
Kir 21 (IK1)
Summary of Ito1 protocolsbull Distribution Atrial PF epicardial M cells gt endocardialbull Pharmacology A common finding in cardiac disease is reduction in Ito1 density with a loss
of heterogeneity of Ito expression and associated action potential prolongation
bull Channel sources1 Transfected cell lines CHO L cell COS-7 cell HEK 293 and Xenopus Oocyte (818 = 44)2 Myocyte (56) human AM (3) rat VM (4) Canine VM 92) and rabbit VM (1)
Guinea Pig (0 very low expression of Ito)
bull Temperature ndash RT = 1215 (80)ndash near or real physiological 315 (20)
1 Pulse sequencesndash Holding -90 to -40 (mode -80 mV) ndash Activation 0 to 75 mV (mode 50 mV)ndash Duration of activation 150 ndash 500 ms (mode 300 ms)
bull Ito measurement peak current or after 70ms of activation
Pulse sequences Ito
Ito (Kir 43)
ID ChannelCurrent
Speciescells CPD Temp Vh(mV)
Prepulse(ms mV)
Activation(ms mV)
Voltage -Dependence(ms mV)
Intervalfrequency
47 hKv43hKChIP22 CHO Methadone RT -90 300 40
49 rKv43 MouseL cell Vanoxerine RT -80 300 20 15s
50 hKv43 HEK293 Alfuzosin RT -80 300 20 15s
206 Ito RatVM Zacopride 24 -40 na 500 -40 to 80 (10)
210 Ito HumanAM Cyamemazine
32-34 -50 500 60
214 Kv43 MonkeyCOS7 Pentamidine na -80 na 500 -40 to 60
215 Ito CanineVM Pioglitazone na -80 5ms -40 na 50
220 Ito RatVM Ajmaline RT -75 300 60
221 rKv42 rKv43 HEK293 RSD1235 25 -80 Brief -50 250 60 1Hz
222 Ito RatVM FK-506 22-24 -80 na 50 1000 -50 to 70 01Hz
224 hKv43 HEK293 Allitridi 22-23 -80 300 60 300 -80 to 60
245 Ito HumanAM DPO RT -80 150 40
256 Ito RabbitVM Carvedilol 34 -60 300 40 300 -50 to 60
263 Kv43 CHO (transient) Spironolactone canrenoic acid
RT -80 250 50 250 -90 to 50 10s
267 Ito ratVM Ebastine na -90 na 75 na -30 to 75
273 Ito CanineVM Thymol 37 -80 5ms- 40 400 50 400 -10 to 60
281 rKv14 Xenopus Oocyte Haloperidol 22 -80 300 0
283 Ito HumanAM Clotrimaxole 21-22 -50 300 50 300 -40 to 60 02Hz
VM= ventricular myocyte AM= Atrial myocyte
Kinetics Pulse Sequences Ito
ID channel cells kinetics Vh (mV)
1st (ms mv)
2nd (ms mV)
3rd (ms mv)
4th (ms mV)
224 hKv43 HEK293 Inactivation -80 -10 to 60
Channel availability -80 na -100 to 40 300 50
Activation conductance -80 8 na na
Recovery time from inactivation -80 30s 50 t -80 300 50 -80
Close state inactivation -90 300 50 3000 -90 t -50 300 50
283 Ito HumanAM Voltage dependence of activation and inactivation
-80 250 -100 to 30 (10) na 50
Recovery time from inactivation -80 30 -40 300 50 t -80 300 50
Extracellular buffer solution of ItoID NaCl NMDG KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-
acetate Blocker pH pH buffer
47 49 50 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 20 033 CdCl2 (01 CaL) BaCl2 (02 Ik1) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 CdCl2
214 140 4 2 1 10 74 NaOH
215 144 56 25 12 5 11 74 na
220 135 54 09 09 10 10 033 CoCl2 (2) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
222 135 4 18 1 2 11 TTX (001)Co++ (2) 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245 132 4 12 10 111 Nisoldipine (04 uM) na na
256 143 54 18 05 5 56 025 Nisoldipine (0003) or CoCl2 (2) and TTX (001) 74 NaOH
263-1 130 4 1 1 10 10 74 NaOH
263-2 140 4 1 1 10 10 TTX (003) CoCl2 (2) 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 140 5 18 1 6 6 Nifedipine (0005) 74 NaOH
283 140 5 1 5 10 033 BaCl2 (01) CdCl2 (02) 74 NaOH
NMDG N-methyl-D-glucamine 263-1 CHO cells 263-2 mouse VM
Pipette solution of ItoID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phospho creatinne
NaCl CaCl2 KOH KH2PO4 pH pH buffer
47 49 50
130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
214 110 142 10 4 10 517 72 KOH
215 45 100 1 10 na 5 72
220 25 130 1 1 5 01 5 3 725 KOH
221 130 1 5 4 01 10 72 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245 20 110 5 5 10 72
256 80 40 10 5 5 5 065 60 72
263 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
Summary of IKur (Kv1531) protocolsbull Distribution a major repolarizing current in human atria but seems to play no
role in the ventricle bull Pharmacology selective IKur blockers to restore and maintain sinus rhythm
without a risk of ventricular pro-arrhythmia
bull Channel sources1 Transfected cell lines CHO L cells HEK 293 and Xenopus Oocyte (710 =
70)2 Myocyte (30) species ndash human AM and mouse VM
bull Temperature RT = 100bull Pulse sequences
ndash Holding range -80 to -50 mV (mode -80 mV)ndash Activation 20 - 60 mV (mode 50 mV)ndash Duration of activation 150 ndash 1000 ms (mode 300 ms)
1 IKur measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKurID Channel
CurrentSpeciescells CPD Temp Vh
(mV)Prepulse Activation
(ms mV)Tail current
Voltage -Dependence(ms mV)
Interval (s)
49 hKv15 MouseL cell vanoxerine RT -80 300 20 10s
221 Kv15 HEK293 RSD1235 25 -80 200 60
224 hKv15 HEK293 Allitridi 22-23 -80 300 50 300 -40 to 60
245-1 hKv15 CHO DPO RT -80 150 40 na -30
245-2 Ikur HumanAM DPO RT -50 300 40 na -30
245-3 hKv15 XO DPO na -80 na 40
262 hKv15 CHO SSR149744C RT -70 1000 30 20s
263 hKv15 Mouse LtK- Spironolactonecanrenoic acid
RT -80 500 60 500 -40 500 -80 to 60
263 Ikur MouseVM Spironolactonecanrenoic acid
RT -80 200 40 500 50 10s
283 Ikur HumanAM clotrimazole 21-22 -50 100 40 150 50 na -30 150 -40 to 50
Ikur (Kv15)
Extracellular buffer solution of IKurID NaCl NMCG K-
gluconate KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-acetate Blocker pH pH
buffer
49 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 10 033 CdCl2 (01) BaCl2 (02) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245-1 132 4 18 12 10 111 72 na
245-2 132 4 12 10 111 nisoldipine (04)
245-3 116 4 10 725
262 140 54 2 2 15 10 74 NaOH
263 130 4 1 1 10 10 74 NaOH
281 96 2 18 1 5 74 NaOH
283 140 5 1 1 5 10 033BaCl2 (02 Ik1) CdCl2 (Ica)
74 NaOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Pipette solution of IKurID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phosphocreatinne
NaCl CaCl2 K-gluconate KH2PO4 pH pH
buffer
49 130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
221 130 1 5 4 01 10 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245-1 15 32 5 5 5 119 735
245-2 20 110 5 5 10 72
245-3 4 10 003 116 725
262 140 2 11 10 2 73 KOH
263 42 80 5 5 5 3 10 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Summary of Iks protocolsbull Distribution all cell types but less in midmyocardial myocytes
bull Pharmacology Atrial and ventricular repolarizationHeart rate dependentbull subunit KCNQ1 (channel forming) Kv71 KvLQT1bull subunit KCNE1 (accessary unit activation of KCNQ1) minK (minimum potassium
subunit)
bull Channel sources1 transfected cell lines CHO HEK 293 Cos-7 cells and Xenopus Oocyte (2136 = 58)2 Myocyte (42) species ndash Canine VM Guinea pig VM rabbit VM and mouse VM (mode
GP)
bull Temperature RT = 1629 55 physiological temperature = 45
bull Pulse sequencesndash Holding potential range = -90 to -30 (mode -80)ndash Depolarization potential 20-60 mV ndash Duration of depolarization 50 ms ndash 10s (mode 2s)ndash Repolarization potential -80 to -10 mV (mode -40 mV)ndash Duration of repolarization 05 ndash 5s (mode 2s)
bull IKs measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation (s mV)
deactivation(s mV)
Interval (s)
47 hKvLQT1hminK CHO RT -80 2 40 01 Hz
49 hKv711hKCNE1 CHO RT -80 2 20 05 -40 15s
50 hKv711hKCNE1 HEk293 RT -80 2 20 05 -40 15s
184 IKs canineepiardial cells 37 -50 2 40 na -30 na
214 Kv711minK HEK293 na -80 5 -60 to 60 5s -50 na
215 IKs caninemyocyte na -40 5 50
224 hKCNQ1hKCNE1 HEK293 22-23 -80 3 -60 to 60 (20) na -50 na
229 hCNE1-hKCNQ1 Transgenic mousemyocyte RT -75 20 -40(Na block) 2 60 2 -40 006 Hz
230 KCNQ1KCNE1 XenopusOocyte 20-22 -80 5 30 na -70
232 KCNQ1KCNE1 HEK293 36 -80 2 -80 to 110 (10) 2 -60 6s
233 KCNQ1KCNE1 CHO na -80 80 -90 3 20 1 -50 005Hz
237 KCNQ1 CHO na -80 na -80 to 60 1 -40
239 KCNQ1KCNE1 HEK293 RT -70 1 -70 to 60 na -40
241 KCNQ1 andor minK MouseCOS-7 22-25 -80 3 -120 to 60 (20) 2 -30
242 hKvLQT1hminK HEK293 19-36 -80 3 -50 to 60 (10) 5 -50
242 hKvLQT1hminK HEK293 19-36 -50 1 or 2 50 na -50 10s
244 KCNQ1 andor KCNE1 CHO na -90 3 -70 to 70 (10) na -60
245 IKs GPMyocyte 35 -50 1 50
249 KCNQ1KCNE1 HEK293 33-34 -40 1 -40 to 80 (10) na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation(s mV)
Voltage dependent(s mV)
Deactivation(s mV)
Interval (s)
251 IKs GPMyocyte 22-2535-37 -40 2 60 2 -40 to 80 (20) na -40
252 IKs GPMyocyte 35 -80 200 -50 3 60 2 -40 60s
252 hminK XenopusOocyte 35 -80 10 20 10 -40 to 60 2 -80
256 IKs Rabbitmyocyte 34 -50 3 -40 to 50 na
260 hKCNQ1hKCNE1 HEK293 RT -80 25 -70 to 100 (10) 25 -40 15s
262 IKs GPMyocyte 35 -80 1 40 5 -10
263 Kv71 and minK CHO RT -80 2 40 na -40
264 IKs GPmyocyte na -80 200 -40 05 50 na -40
267 IKs GPmyocyte na -90 na 0 na 40 01- 015 0-80 -90
273 IKs Caninemyocyte 37 -40 3 50 na -40
274 IKs GPmyocyte 36 -30 05 50 or 2 70 05 (2) -30 to 70 na -30
275 KCNQ1KCNE1 CHO 21 -80 1 40 01Hz
276 hKCNQ1hminK CHO 22 -80 6 40 na -80 005Hz
278 IKs Canine myocyte 35-37 -50 3 -50 to 50 2s -40
283 KCNQ1KCNE1 HEK293 21-22 -80 3 40 3 -80 to 40 na -40
286 IKs GPmyocyte 34-36 -50 005 30 005 ndash3 -50 to 30 na -50
287 Iks GPmyocyte 36 -80 na -40 2 60 2 -40 to 80 na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
47 49 50
137 4 18 1 10 10 74 NaOH
184 145 4 18 18 10 11 NaH2PO4 1 E4031 (5uM) Cadmium (300 uM) 74 NaOH
214 140 4 2 1 10 74 NaOH
215 144 5 56 25 5 Dextrose 11 Nifedipine (5 uM) E4031 (1 uM) 74 NaOH
224 140 54 18 1 10 10 NaH2PO4 033 73 NaOH
229 132 48 2 12 10 5 E4031 nisoldipine 74 NaOH
230 96 2 18 1 5 74 NaOH
232 140 54 18 1 5 55 74 NaOH
233 17 120 4 2 1 10 10 74 NaOH
237 130 5 18 1 10 Na-Acetate 28 74 NaOH
239 150 54 18 4 10 74 NaOH
241 135 5 25 12 5 10 74 NaOH
242 132 4 18 12 10 111 74 NaOH
244 140 4 18 12 55 11 74 NaOH
249 140 54 18 05 5 55 NaH2PO4 033 74 NaOH
250 140 54 18 1 5 10 74 NaOH
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
251 1 1 2 10 N-methly-d-glucamine 132
lanthanum chloride 005 mMcadmium chloride 02 mM 74 HCl
252 140 47 13 1 10 10 74 NaOH
256 09 5 N-methly-d-glucamine 149
Nisoldipine 3 uME4031 10 uM 74 HCl
260 140 54 18 1 5 10 NaH2PO4 03 73 NaOH
262 137 4 18 2 10 115 Pyrubic acid 5 Thiamine 01 74 NaOH
263 130 4 1 1 10 10 TTX 003 mM CoCl2 2 mM 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10 KH2PO4 12 MgSO4 12 NaHCO3 44
Nifetidine 5 uM 4-aminopyridine 3 mM 72 NaOH
274 140 1 5 10 Cd2+ 02 mM 74 NaOH
275 130 54 18 1 10 5 74 NaOH
278 136 54 1 1 5 Dextrose 10 NaH2PO4 033 E4031 5 uM 74 NaOH
283 140 5 1 1 5 10 NaH2PO4 033 74 NaOH
286 09 5 5 N-methly-d-glucamine 149
Nisoldipine 04 uME4031 5 uM 74 NaOH
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
49 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
214 110 142 10 4 4 517 72 KOH
215 45 100 1 10 na 5 72
224 20 110 1 5 5 01 10 5 72 KOH
229 110 11 5 10 55 73
230 3000
232 20 125 10 10
233 140 1 1 20 73 KOH
237 10 125 1 10 5 10 10 72 KOH
239 30 130 01 4 10 74
241 117 2 11 11 10 1 72
242 15 119 32 5 5 5 74 KOH
244 130 1 5 5 10 74 KOH
249 130 5 5 01 5 5 74 HCl
250 80 40 10 5 01 5 40 065 72 HCl
251 50 60 1 11 5 10 1 74 KOH
252 140 11 1 10 10 72 KOH
amphotericin-B
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
256 80 40 10 5 5 5 065 72
262 150 1 5 4 5 5 73 KOH
263^ 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
274 40 106 1 5 4 5 72 KOH
276 20 110 1 10 5 10 10 1 72 KOH
278 20 110 1 10 5 01 10 5 72 KOH
283 20 110 1 5 5 01 1 5 72 KOH
286 50 1 5 5 01 5 2 10 72 KOH
^phosphatidylinositol 45-diphosphate (5 mgml 1) was added to the internal solution to avoid the rundown of the current 70 L-aspartic acid 70 KOH
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Late sodium channel protocolsRef ID Year Drug Holding (mV) Late Na (ms) Late Na (mV) Peak Na
(ms mV)Frequency
(Hz)
173 1997 -120 100 -20 same 02
100 1998 -150 720 -20 24 -90 to 30
174 1998 R56865 -100 350 -20 No data 02
177 2000 Flecainide -150 Mean (20 and 235) -20 same 5
179 2000 BDF 9198 -140 250 -30 No info
178 2001 Amiodarone -160 200-220 -30 No time info 01
180 2002 -140 750 -40 25 -30
186 2006 -140 2000 na No info 01
205 2009 Ranolazine -140 Mean (46 and 48) -20 24 -20
199 2012 -120 300 -20 No info
160 2013 Ranolazine -130 50 -10 20 -10
195 2013 GS967 -120 220 -20 20 -20
Ex) Ref ID 195peak late
-120 mV complete removal of inactivation
-20 mV
INa enhancer
Pulse20 vs 220 ms
Cardiac potassium channels
ID248 name clone Channel blocker role note
Ito1 Transient outward Kv43Kv14 4-AP (closed state)bullEarly rapid repolarization (Phase I)bullDetermine the height of early plateau
Ito2 is not found in cardiac cells
IKur Ultra rapid delayed rectifier Kv15 4-AP (selective) bullAtrial repolarization
IKs Slow delayed rectifier KCNQ1(KvLQT1) KCNE1 (hminK) Chromanol (293B)
bullAtrial and ventricular repolarizationbullHeart rate dependent
Activation ~0 mV
IKr rapid delayed rectifier Kv111 (hERG) E-4031bullRapid activation and inactivation strong inward rectificationbullCardiac APD and refractoriness
Activation ~-30mV[K+] dependent
IK1 Inward rectifier Kir21 Ba2+ (IC50= 20 uM)
bullMaintain the resting membrane potentialbullControl excitabilitybullInitiate the final repolarization phase
Nav15
Kv43 (Ito f) Kv15 (IKur)
KvLQT1minK (IKs)
Cav12 (ICaL) hERG (IKr)
Cav32 (IcaT)
Kir 21 (IK1)
Summary of Ito1 protocolsbull Distribution Atrial PF epicardial M cells gt endocardialbull Pharmacology A common finding in cardiac disease is reduction in Ito1 density with a loss
of heterogeneity of Ito expression and associated action potential prolongation
bull Channel sources1 Transfected cell lines CHO L cell COS-7 cell HEK 293 and Xenopus Oocyte (818 = 44)2 Myocyte (56) human AM (3) rat VM (4) Canine VM 92) and rabbit VM (1)
Guinea Pig (0 very low expression of Ito)
bull Temperature ndash RT = 1215 (80)ndash near or real physiological 315 (20)
1 Pulse sequencesndash Holding -90 to -40 (mode -80 mV) ndash Activation 0 to 75 mV (mode 50 mV)ndash Duration of activation 150 ndash 500 ms (mode 300 ms)
bull Ito measurement peak current or after 70ms of activation
Pulse sequences Ito
Ito (Kir 43)
ID ChannelCurrent
Speciescells CPD Temp Vh(mV)
Prepulse(ms mV)
Activation(ms mV)
Voltage -Dependence(ms mV)
Intervalfrequency
47 hKv43hKChIP22 CHO Methadone RT -90 300 40
49 rKv43 MouseL cell Vanoxerine RT -80 300 20 15s
50 hKv43 HEK293 Alfuzosin RT -80 300 20 15s
206 Ito RatVM Zacopride 24 -40 na 500 -40 to 80 (10)
210 Ito HumanAM Cyamemazine
32-34 -50 500 60
214 Kv43 MonkeyCOS7 Pentamidine na -80 na 500 -40 to 60
215 Ito CanineVM Pioglitazone na -80 5ms -40 na 50
220 Ito RatVM Ajmaline RT -75 300 60
221 rKv42 rKv43 HEK293 RSD1235 25 -80 Brief -50 250 60 1Hz
222 Ito RatVM FK-506 22-24 -80 na 50 1000 -50 to 70 01Hz
224 hKv43 HEK293 Allitridi 22-23 -80 300 60 300 -80 to 60
245 Ito HumanAM DPO RT -80 150 40
256 Ito RabbitVM Carvedilol 34 -60 300 40 300 -50 to 60
263 Kv43 CHO (transient) Spironolactone canrenoic acid
RT -80 250 50 250 -90 to 50 10s
267 Ito ratVM Ebastine na -90 na 75 na -30 to 75
273 Ito CanineVM Thymol 37 -80 5ms- 40 400 50 400 -10 to 60
281 rKv14 Xenopus Oocyte Haloperidol 22 -80 300 0
283 Ito HumanAM Clotrimaxole 21-22 -50 300 50 300 -40 to 60 02Hz
VM= ventricular myocyte AM= Atrial myocyte
Kinetics Pulse Sequences Ito
ID channel cells kinetics Vh (mV)
1st (ms mv)
2nd (ms mV)
3rd (ms mv)
4th (ms mV)
224 hKv43 HEK293 Inactivation -80 -10 to 60
Channel availability -80 na -100 to 40 300 50
Activation conductance -80 8 na na
Recovery time from inactivation -80 30s 50 t -80 300 50 -80
Close state inactivation -90 300 50 3000 -90 t -50 300 50
283 Ito HumanAM Voltage dependence of activation and inactivation
-80 250 -100 to 30 (10) na 50
Recovery time from inactivation -80 30 -40 300 50 t -80 300 50
Extracellular buffer solution of ItoID NaCl NMDG KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-
acetate Blocker pH pH buffer
47 49 50 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 20 033 CdCl2 (01 CaL) BaCl2 (02 Ik1) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 CdCl2
214 140 4 2 1 10 74 NaOH
215 144 56 25 12 5 11 74 na
220 135 54 09 09 10 10 033 CoCl2 (2) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
222 135 4 18 1 2 11 TTX (001)Co++ (2) 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245 132 4 12 10 111 Nisoldipine (04 uM) na na
256 143 54 18 05 5 56 025 Nisoldipine (0003) or CoCl2 (2) and TTX (001) 74 NaOH
263-1 130 4 1 1 10 10 74 NaOH
263-2 140 4 1 1 10 10 TTX (003) CoCl2 (2) 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 140 5 18 1 6 6 Nifedipine (0005) 74 NaOH
283 140 5 1 5 10 033 BaCl2 (01) CdCl2 (02) 74 NaOH
NMDG N-methyl-D-glucamine 263-1 CHO cells 263-2 mouse VM
Pipette solution of ItoID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phospho creatinne
NaCl CaCl2 KOH KH2PO4 pH pH buffer
47 49 50
130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
214 110 142 10 4 10 517 72 KOH
215 45 100 1 10 na 5 72
220 25 130 1 1 5 01 5 3 725 KOH
221 130 1 5 4 01 10 72 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245 20 110 5 5 10 72
256 80 40 10 5 5 5 065 60 72
263 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
Summary of IKur (Kv1531) protocolsbull Distribution a major repolarizing current in human atria but seems to play no
role in the ventricle bull Pharmacology selective IKur blockers to restore and maintain sinus rhythm
without a risk of ventricular pro-arrhythmia
bull Channel sources1 Transfected cell lines CHO L cells HEK 293 and Xenopus Oocyte (710 =
70)2 Myocyte (30) species ndash human AM and mouse VM
bull Temperature RT = 100bull Pulse sequences
ndash Holding range -80 to -50 mV (mode -80 mV)ndash Activation 20 - 60 mV (mode 50 mV)ndash Duration of activation 150 ndash 1000 ms (mode 300 ms)
1 IKur measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKurID Channel
CurrentSpeciescells CPD Temp Vh
(mV)Prepulse Activation
(ms mV)Tail current
Voltage -Dependence(ms mV)
Interval (s)
49 hKv15 MouseL cell vanoxerine RT -80 300 20 10s
221 Kv15 HEK293 RSD1235 25 -80 200 60
224 hKv15 HEK293 Allitridi 22-23 -80 300 50 300 -40 to 60
245-1 hKv15 CHO DPO RT -80 150 40 na -30
245-2 Ikur HumanAM DPO RT -50 300 40 na -30
245-3 hKv15 XO DPO na -80 na 40
262 hKv15 CHO SSR149744C RT -70 1000 30 20s
263 hKv15 Mouse LtK- Spironolactonecanrenoic acid
RT -80 500 60 500 -40 500 -80 to 60
263 Ikur MouseVM Spironolactonecanrenoic acid
RT -80 200 40 500 50 10s
283 Ikur HumanAM clotrimazole 21-22 -50 100 40 150 50 na -30 150 -40 to 50
Ikur (Kv15)
Extracellular buffer solution of IKurID NaCl NMCG K-
gluconate KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-acetate Blocker pH pH
buffer
49 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 10 033 CdCl2 (01) BaCl2 (02) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245-1 132 4 18 12 10 111 72 na
245-2 132 4 12 10 111 nisoldipine (04)
245-3 116 4 10 725
262 140 54 2 2 15 10 74 NaOH
263 130 4 1 1 10 10 74 NaOH
281 96 2 18 1 5 74 NaOH
283 140 5 1 1 5 10 033BaCl2 (02 Ik1) CdCl2 (Ica)
74 NaOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Pipette solution of IKurID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phosphocreatinne
NaCl CaCl2 K-gluconate KH2PO4 pH pH
buffer
49 130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
221 130 1 5 4 01 10 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245-1 15 32 5 5 5 119 735
245-2 20 110 5 5 10 72
245-3 4 10 003 116 725
262 140 2 11 10 2 73 KOH
263 42 80 5 5 5 3 10 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Summary of Iks protocolsbull Distribution all cell types but less in midmyocardial myocytes
bull Pharmacology Atrial and ventricular repolarizationHeart rate dependentbull subunit KCNQ1 (channel forming) Kv71 KvLQT1bull subunit KCNE1 (accessary unit activation of KCNQ1) minK (minimum potassium
subunit)
bull Channel sources1 transfected cell lines CHO HEK 293 Cos-7 cells and Xenopus Oocyte (2136 = 58)2 Myocyte (42) species ndash Canine VM Guinea pig VM rabbit VM and mouse VM (mode
GP)
bull Temperature RT = 1629 55 physiological temperature = 45
bull Pulse sequencesndash Holding potential range = -90 to -30 (mode -80)ndash Depolarization potential 20-60 mV ndash Duration of depolarization 50 ms ndash 10s (mode 2s)ndash Repolarization potential -80 to -10 mV (mode -40 mV)ndash Duration of repolarization 05 ndash 5s (mode 2s)
bull IKs measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation (s mV)
deactivation(s mV)
Interval (s)
47 hKvLQT1hminK CHO RT -80 2 40 01 Hz
49 hKv711hKCNE1 CHO RT -80 2 20 05 -40 15s
50 hKv711hKCNE1 HEk293 RT -80 2 20 05 -40 15s
184 IKs canineepiardial cells 37 -50 2 40 na -30 na
214 Kv711minK HEK293 na -80 5 -60 to 60 5s -50 na
215 IKs caninemyocyte na -40 5 50
224 hKCNQ1hKCNE1 HEK293 22-23 -80 3 -60 to 60 (20) na -50 na
229 hCNE1-hKCNQ1 Transgenic mousemyocyte RT -75 20 -40(Na block) 2 60 2 -40 006 Hz
230 KCNQ1KCNE1 XenopusOocyte 20-22 -80 5 30 na -70
232 KCNQ1KCNE1 HEK293 36 -80 2 -80 to 110 (10) 2 -60 6s
233 KCNQ1KCNE1 CHO na -80 80 -90 3 20 1 -50 005Hz
237 KCNQ1 CHO na -80 na -80 to 60 1 -40
239 KCNQ1KCNE1 HEK293 RT -70 1 -70 to 60 na -40
241 KCNQ1 andor minK MouseCOS-7 22-25 -80 3 -120 to 60 (20) 2 -30
242 hKvLQT1hminK HEK293 19-36 -80 3 -50 to 60 (10) 5 -50
242 hKvLQT1hminK HEK293 19-36 -50 1 or 2 50 na -50 10s
244 KCNQ1 andor KCNE1 CHO na -90 3 -70 to 70 (10) na -60
245 IKs GPMyocyte 35 -50 1 50
249 KCNQ1KCNE1 HEK293 33-34 -40 1 -40 to 80 (10) na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation(s mV)
Voltage dependent(s mV)
Deactivation(s mV)
Interval (s)
251 IKs GPMyocyte 22-2535-37 -40 2 60 2 -40 to 80 (20) na -40
252 IKs GPMyocyte 35 -80 200 -50 3 60 2 -40 60s
252 hminK XenopusOocyte 35 -80 10 20 10 -40 to 60 2 -80
256 IKs Rabbitmyocyte 34 -50 3 -40 to 50 na
260 hKCNQ1hKCNE1 HEK293 RT -80 25 -70 to 100 (10) 25 -40 15s
262 IKs GPMyocyte 35 -80 1 40 5 -10
263 Kv71 and minK CHO RT -80 2 40 na -40
264 IKs GPmyocyte na -80 200 -40 05 50 na -40
267 IKs GPmyocyte na -90 na 0 na 40 01- 015 0-80 -90
273 IKs Caninemyocyte 37 -40 3 50 na -40
274 IKs GPmyocyte 36 -30 05 50 or 2 70 05 (2) -30 to 70 na -30
275 KCNQ1KCNE1 CHO 21 -80 1 40 01Hz
276 hKCNQ1hminK CHO 22 -80 6 40 na -80 005Hz
278 IKs Canine myocyte 35-37 -50 3 -50 to 50 2s -40
283 KCNQ1KCNE1 HEK293 21-22 -80 3 40 3 -80 to 40 na -40
286 IKs GPmyocyte 34-36 -50 005 30 005 ndash3 -50 to 30 na -50
287 Iks GPmyocyte 36 -80 na -40 2 60 2 -40 to 80 na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
47 49 50
137 4 18 1 10 10 74 NaOH
184 145 4 18 18 10 11 NaH2PO4 1 E4031 (5uM) Cadmium (300 uM) 74 NaOH
214 140 4 2 1 10 74 NaOH
215 144 5 56 25 5 Dextrose 11 Nifedipine (5 uM) E4031 (1 uM) 74 NaOH
224 140 54 18 1 10 10 NaH2PO4 033 73 NaOH
229 132 48 2 12 10 5 E4031 nisoldipine 74 NaOH
230 96 2 18 1 5 74 NaOH
232 140 54 18 1 5 55 74 NaOH
233 17 120 4 2 1 10 10 74 NaOH
237 130 5 18 1 10 Na-Acetate 28 74 NaOH
239 150 54 18 4 10 74 NaOH
241 135 5 25 12 5 10 74 NaOH
242 132 4 18 12 10 111 74 NaOH
244 140 4 18 12 55 11 74 NaOH
249 140 54 18 05 5 55 NaH2PO4 033 74 NaOH
250 140 54 18 1 5 10 74 NaOH
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
251 1 1 2 10 N-methly-d-glucamine 132
lanthanum chloride 005 mMcadmium chloride 02 mM 74 HCl
252 140 47 13 1 10 10 74 NaOH
256 09 5 N-methly-d-glucamine 149
Nisoldipine 3 uME4031 10 uM 74 HCl
260 140 54 18 1 5 10 NaH2PO4 03 73 NaOH
262 137 4 18 2 10 115 Pyrubic acid 5 Thiamine 01 74 NaOH
263 130 4 1 1 10 10 TTX 003 mM CoCl2 2 mM 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10 KH2PO4 12 MgSO4 12 NaHCO3 44
Nifetidine 5 uM 4-aminopyridine 3 mM 72 NaOH
274 140 1 5 10 Cd2+ 02 mM 74 NaOH
275 130 54 18 1 10 5 74 NaOH
278 136 54 1 1 5 Dextrose 10 NaH2PO4 033 E4031 5 uM 74 NaOH
283 140 5 1 1 5 10 NaH2PO4 033 74 NaOH
286 09 5 5 N-methly-d-glucamine 149
Nisoldipine 04 uME4031 5 uM 74 NaOH
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
49 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
214 110 142 10 4 4 517 72 KOH
215 45 100 1 10 na 5 72
224 20 110 1 5 5 01 10 5 72 KOH
229 110 11 5 10 55 73
230 3000
232 20 125 10 10
233 140 1 1 20 73 KOH
237 10 125 1 10 5 10 10 72 KOH
239 30 130 01 4 10 74
241 117 2 11 11 10 1 72
242 15 119 32 5 5 5 74 KOH
244 130 1 5 5 10 74 KOH
249 130 5 5 01 5 5 74 HCl
250 80 40 10 5 01 5 40 065 72 HCl
251 50 60 1 11 5 10 1 74 KOH
252 140 11 1 10 10 72 KOH
amphotericin-B
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
256 80 40 10 5 5 5 065 72
262 150 1 5 4 5 5 73 KOH
263^ 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
274 40 106 1 5 4 5 72 KOH
276 20 110 1 10 5 10 10 1 72 KOH
278 20 110 1 10 5 01 10 5 72 KOH
283 20 110 1 5 5 01 1 5 72 KOH
286 50 1 5 5 01 5 2 10 72 KOH
^phosphatidylinositol 45-diphosphate (5 mgml 1) was added to the internal solution to avoid the rundown of the current 70 L-aspartic acid 70 KOH
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Cardiac potassium channels
ID248 name clone Channel blocker role note
Ito1 Transient outward Kv43Kv14 4-AP (closed state)bullEarly rapid repolarization (Phase I)bullDetermine the height of early plateau
Ito2 is not found in cardiac cells
IKur Ultra rapid delayed rectifier Kv15 4-AP (selective) bullAtrial repolarization
IKs Slow delayed rectifier KCNQ1(KvLQT1) KCNE1 (hminK) Chromanol (293B)
bullAtrial and ventricular repolarizationbullHeart rate dependent
Activation ~0 mV
IKr rapid delayed rectifier Kv111 (hERG) E-4031bullRapid activation and inactivation strong inward rectificationbullCardiac APD and refractoriness
Activation ~-30mV[K+] dependent
IK1 Inward rectifier Kir21 Ba2+ (IC50= 20 uM)
bullMaintain the resting membrane potentialbullControl excitabilitybullInitiate the final repolarization phase
Nav15
Kv43 (Ito f) Kv15 (IKur)
KvLQT1minK (IKs)
Cav12 (ICaL) hERG (IKr)
Cav32 (IcaT)
Kir 21 (IK1)
Summary of Ito1 protocolsbull Distribution Atrial PF epicardial M cells gt endocardialbull Pharmacology A common finding in cardiac disease is reduction in Ito1 density with a loss
of heterogeneity of Ito expression and associated action potential prolongation
bull Channel sources1 Transfected cell lines CHO L cell COS-7 cell HEK 293 and Xenopus Oocyte (818 = 44)2 Myocyte (56) human AM (3) rat VM (4) Canine VM 92) and rabbit VM (1)
Guinea Pig (0 very low expression of Ito)
bull Temperature ndash RT = 1215 (80)ndash near or real physiological 315 (20)
1 Pulse sequencesndash Holding -90 to -40 (mode -80 mV) ndash Activation 0 to 75 mV (mode 50 mV)ndash Duration of activation 150 ndash 500 ms (mode 300 ms)
bull Ito measurement peak current or after 70ms of activation
Pulse sequences Ito
Ito (Kir 43)
ID ChannelCurrent
Speciescells CPD Temp Vh(mV)
Prepulse(ms mV)
Activation(ms mV)
Voltage -Dependence(ms mV)
Intervalfrequency
47 hKv43hKChIP22 CHO Methadone RT -90 300 40
49 rKv43 MouseL cell Vanoxerine RT -80 300 20 15s
50 hKv43 HEK293 Alfuzosin RT -80 300 20 15s
206 Ito RatVM Zacopride 24 -40 na 500 -40 to 80 (10)
210 Ito HumanAM Cyamemazine
32-34 -50 500 60
214 Kv43 MonkeyCOS7 Pentamidine na -80 na 500 -40 to 60
215 Ito CanineVM Pioglitazone na -80 5ms -40 na 50
220 Ito RatVM Ajmaline RT -75 300 60
221 rKv42 rKv43 HEK293 RSD1235 25 -80 Brief -50 250 60 1Hz
222 Ito RatVM FK-506 22-24 -80 na 50 1000 -50 to 70 01Hz
224 hKv43 HEK293 Allitridi 22-23 -80 300 60 300 -80 to 60
245 Ito HumanAM DPO RT -80 150 40
256 Ito RabbitVM Carvedilol 34 -60 300 40 300 -50 to 60
263 Kv43 CHO (transient) Spironolactone canrenoic acid
RT -80 250 50 250 -90 to 50 10s
267 Ito ratVM Ebastine na -90 na 75 na -30 to 75
273 Ito CanineVM Thymol 37 -80 5ms- 40 400 50 400 -10 to 60
281 rKv14 Xenopus Oocyte Haloperidol 22 -80 300 0
283 Ito HumanAM Clotrimaxole 21-22 -50 300 50 300 -40 to 60 02Hz
VM= ventricular myocyte AM= Atrial myocyte
Kinetics Pulse Sequences Ito
ID channel cells kinetics Vh (mV)
1st (ms mv)
2nd (ms mV)
3rd (ms mv)
4th (ms mV)
224 hKv43 HEK293 Inactivation -80 -10 to 60
Channel availability -80 na -100 to 40 300 50
Activation conductance -80 8 na na
Recovery time from inactivation -80 30s 50 t -80 300 50 -80
Close state inactivation -90 300 50 3000 -90 t -50 300 50
283 Ito HumanAM Voltage dependence of activation and inactivation
-80 250 -100 to 30 (10) na 50
Recovery time from inactivation -80 30 -40 300 50 t -80 300 50
Extracellular buffer solution of ItoID NaCl NMDG KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-
acetate Blocker pH pH buffer
47 49 50 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 20 033 CdCl2 (01 CaL) BaCl2 (02 Ik1) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 CdCl2
214 140 4 2 1 10 74 NaOH
215 144 56 25 12 5 11 74 na
220 135 54 09 09 10 10 033 CoCl2 (2) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
222 135 4 18 1 2 11 TTX (001)Co++ (2) 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245 132 4 12 10 111 Nisoldipine (04 uM) na na
256 143 54 18 05 5 56 025 Nisoldipine (0003) or CoCl2 (2) and TTX (001) 74 NaOH
263-1 130 4 1 1 10 10 74 NaOH
263-2 140 4 1 1 10 10 TTX (003) CoCl2 (2) 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 140 5 18 1 6 6 Nifedipine (0005) 74 NaOH
283 140 5 1 5 10 033 BaCl2 (01) CdCl2 (02) 74 NaOH
NMDG N-methyl-D-glucamine 263-1 CHO cells 263-2 mouse VM
Pipette solution of ItoID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phospho creatinne
NaCl CaCl2 KOH KH2PO4 pH pH buffer
47 49 50
130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
214 110 142 10 4 10 517 72 KOH
215 45 100 1 10 na 5 72
220 25 130 1 1 5 01 5 3 725 KOH
221 130 1 5 4 01 10 72 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245 20 110 5 5 10 72
256 80 40 10 5 5 5 065 60 72
263 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
Summary of IKur (Kv1531) protocolsbull Distribution a major repolarizing current in human atria but seems to play no
role in the ventricle bull Pharmacology selective IKur blockers to restore and maintain sinus rhythm
without a risk of ventricular pro-arrhythmia
bull Channel sources1 Transfected cell lines CHO L cells HEK 293 and Xenopus Oocyte (710 =
70)2 Myocyte (30) species ndash human AM and mouse VM
bull Temperature RT = 100bull Pulse sequences
ndash Holding range -80 to -50 mV (mode -80 mV)ndash Activation 20 - 60 mV (mode 50 mV)ndash Duration of activation 150 ndash 1000 ms (mode 300 ms)
1 IKur measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKurID Channel
CurrentSpeciescells CPD Temp Vh
(mV)Prepulse Activation
(ms mV)Tail current
Voltage -Dependence(ms mV)
Interval (s)
49 hKv15 MouseL cell vanoxerine RT -80 300 20 10s
221 Kv15 HEK293 RSD1235 25 -80 200 60
224 hKv15 HEK293 Allitridi 22-23 -80 300 50 300 -40 to 60
245-1 hKv15 CHO DPO RT -80 150 40 na -30
245-2 Ikur HumanAM DPO RT -50 300 40 na -30
245-3 hKv15 XO DPO na -80 na 40
262 hKv15 CHO SSR149744C RT -70 1000 30 20s
263 hKv15 Mouse LtK- Spironolactonecanrenoic acid
RT -80 500 60 500 -40 500 -80 to 60
263 Ikur MouseVM Spironolactonecanrenoic acid
RT -80 200 40 500 50 10s
283 Ikur HumanAM clotrimazole 21-22 -50 100 40 150 50 na -30 150 -40 to 50
Ikur (Kv15)
Extracellular buffer solution of IKurID NaCl NMCG K-
gluconate KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-acetate Blocker pH pH
buffer
49 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 10 033 CdCl2 (01) BaCl2 (02) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245-1 132 4 18 12 10 111 72 na
245-2 132 4 12 10 111 nisoldipine (04)
245-3 116 4 10 725
262 140 54 2 2 15 10 74 NaOH
263 130 4 1 1 10 10 74 NaOH
281 96 2 18 1 5 74 NaOH
283 140 5 1 1 5 10 033BaCl2 (02 Ik1) CdCl2 (Ica)
74 NaOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Pipette solution of IKurID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phosphocreatinne
NaCl CaCl2 K-gluconate KH2PO4 pH pH
buffer
49 130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
221 130 1 5 4 01 10 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245-1 15 32 5 5 5 119 735
245-2 20 110 5 5 10 72
245-3 4 10 003 116 725
262 140 2 11 10 2 73 KOH
263 42 80 5 5 5 3 10 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Summary of Iks protocolsbull Distribution all cell types but less in midmyocardial myocytes
bull Pharmacology Atrial and ventricular repolarizationHeart rate dependentbull subunit KCNQ1 (channel forming) Kv71 KvLQT1bull subunit KCNE1 (accessary unit activation of KCNQ1) minK (minimum potassium
subunit)
bull Channel sources1 transfected cell lines CHO HEK 293 Cos-7 cells and Xenopus Oocyte (2136 = 58)2 Myocyte (42) species ndash Canine VM Guinea pig VM rabbit VM and mouse VM (mode
GP)
bull Temperature RT = 1629 55 physiological temperature = 45
bull Pulse sequencesndash Holding potential range = -90 to -30 (mode -80)ndash Depolarization potential 20-60 mV ndash Duration of depolarization 50 ms ndash 10s (mode 2s)ndash Repolarization potential -80 to -10 mV (mode -40 mV)ndash Duration of repolarization 05 ndash 5s (mode 2s)
bull IKs measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation (s mV)
deactivation(s mV)
Interval (s)
47 hKvLQT1hminK CHO RT -80 2 40 01 Hz
49 hKv711hKCNE1 CHO RT -80 2 20 05 -40 15s
50 hKv711hKCNE1 HEk293 RT -80 2 20 05 -40 15s
184 IKs canineepiardial cells 37 -50 2 40 na -30 na
214 Kv711minK HEK293 na -80 5 -60 to 60 5s -50 na
215 IKs caninemyocyte na -40 5 50
224 hKCNQ1hKCNE1 HEK293 22-23 -80 3 -60 to 60 (20) na -50 na
229 hCNE1-hKCNQ1 Transgenic mousemyocyte RT -75 20 -40(Na block) 2 60 2 -40 006 Hz
230 KCNQ1KCNE1 XenopusOocyte 20-22 -80 5 30 na -70
232 KCNQ1KCNE1 HEK293 36 -80 2 -80 to 110 (10) 2 -60 6s
233 KCNQ1KCNE1 CHO na -80 80 -90 3 20 1 -50 005Hz
237 KCNQ1 CHO na -80 na -80 to 60 1 -40
239 KCNQ1KCNE1 HEK293 RT -70 1 -70 to 60 na -40
241 KCNQ1 andor minK MouseCOS-7 22-25 -80 3 -120 to 60 (20) 2 -30
242 hKvLQT1hminK HEK293 19-36 -80 3 -50 to 60 (10) 5 -50
242 hKvLQT1hminK HEK293 19-36 -50 1 or 2 50 na -50 10s
244 KCNQ1 andor KCNE1 CHO na -90 3 -70 to 70 (10) na -60
245 IKs GPMyocyte 35 -50 1 50
249 KCNQ1KCNE1 HEK293 33-34 -40 1 -40 to 80 (10) na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation(s mV)
Voltage dependent(s mV)
Deactivation(s mV)
Interval (s)
251 IKs GPMyocyte 22-2535-37 -40 2 60 2 -40 to 80 (20) na -40
252 IKs GPMyocyte 35 -80 200 -50 3 60 2 -40 60s
252 hminK XenopusOocyte 35 -80 10 20 10 -40 to 60 2 -80
256 IKs Rabbitmyocyte 34 -50 3 -40 to 50 na
260 hKCNQ1hKCNE1 HEK293 RT -80 25 -70 to 100 (10) 25 -40 15s
262 IKs GPMyocyte 35 -80 1 40 5 -10
263 Kv71 and minK CHO RT -80 2 40 na -40
264 IKs GPmyocyte na -80 200 -40 05 50 na -40
267 IKs GPmyocyte na -90 na 0 na 40 01- 015 0-80 -90
273 IKs Caninemyocyte 37 -40 3 50 na -40
274 IKs GPmyocyte 36 -30 05 50 or 2 70 05 (2) -30 to 70 na -30
275 KCNQ1KCNE1 CHO 21 -80 1 40 01Hz
276 hKCNQ1hminK CHO 22 -80 6 40 na -80 005Hz
278 IKs Canine myocyte 35-37 -50 3 -50 to 50 2s -40
283 KCNQ1KCNE1 HEK293 21-22 -80 3 40 3 -80 to 40 na -40
286 IKs GPmyocyte 34-36 -50 005 30 005 ndash3 -50 to 30 na -50
287 Iks GPmyocyte 36 -80 na -40 2 60 2 -40 to 80 na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
47 49 50
137 4 18 1 10 10 74 NaOH
184 145 4 18 18 10 11 NaH2PO4 1 E4031 (5uM) Cadmium (300 uM) 74 NaOH
214 140 4 2 1 10 74 NaOH
215 144 5 56 25 5 Dextrose 11 Nifedipine (5 uM) E4031 (1 uM) 74 NaOH
224 140 54 18 1 10 10 NaH2PO4 033 73 NaOH
229 132 48 2 12 10 5 E4031 nisoldipine 74 NaOH
230 96 2 18 1 5 74 NaOH
232 140 54 18 1 5 55 74 NaOH
233 17 120 4 2 1 10 10 74 NaOH
237 130 5 18 1 10 Na-Acetate 28 74 NaOH
239 150 54 18 4 10 74 NaOH
241 135 5 25 12 5 10 74 NaOH
242 132 4 18 12 10 111 74 NaOH
244 140 4 18 12 55 11 74 NaOH
249 140 54 18 05 5 55 NaH2PO4 033 74 NaOH
250 140 54 18 1 5 10 74 NaOH
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
251 1 1 2 10 N-methly-d-glucamine 132
lanthanum chloride 005 mMcadmium chloride 02 mM 74 HCl
252 140 47 13 1 10 10 74 NaOH
256 09 5 N-methly-d-glucamine 149
Nisoldipine 3 uME4031 10 uM 74 HCl
260 140 54 18 1 5 10 NaH2PO4 03 73 NaOH
262 137 4 18 2 10 115 Pyrubic acid 5 Thiamine 01 74 NaOH
263 130 4 1 1 10 10 TTX 003 mM CoCl2 2 mM 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10 KH2PO4 12 MgSO4 12 NaHCO3 44
Nifetidine 5 uM 4-aminopyridine 3 mM 72 NaOH
274 140 1 5 10 Cd2+ 02 mM 74 NaOH
275 130 54 18 1 10 5 74 NaOH
278 136 54 1 1 5 Dextrose 10 NaH2PO4 033 E4031 5 uM 74 NaOH
283 140 5 1 1 5 10 NaH2PO4 033 74 NaOH
286 09 5 5 N-methly-d-glucamine 149
Nisoldipine 04 uME4031 5 uM 74 NaOH
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
49 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
214 110 142 10 4 4 517 72 KOH
215 45 100 1 10 na 5 72
224 20 110 1 5 5 01 10 5 72 KOH
229 110 11 5 10 55 73
230 3000
232 20 125 10 10
233 140 1 1 20 73 KOH
237 10 125 1 10 5 10 10 72 KOH
239 30 130 01 4 10 74
241 117 2 11 11 10 1 72
242 15 119 32 5 5 5 74 KOH
244 130 1 5 5 10 74 KOH
249 130 5 5 01 5 5 74 HCl
250 80 40 10 5 01 5 40 065 72 HCl
251 50 60 1 11 5 10 1 74 KOH
252 140 11 1 10 10 72 KOH
amphotericin-B
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
256 80 40 10 5 5 5 065 72
262 150 1 5 4 5 5 73 KOH
263^ 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
274 40 106 1 5 4 5 72 KOH
276 20 110 1 10 5 10 10 1 72 KOH
278 20 110 1 10 5 01 10 5 72 KOH
283 20 110 1 5 5 01 1 5 72 KOH
286 50 1 5 5 01 5 2 10 72 KOH
^phosphatidylinositol 45-diphosphate (5 mgml 1) was added to the internal solution to avoid the rundown of the current 70 L-aspartic acid 70 KOH
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Summary of Ito1 protocolsbull Distribution Atrial PF epicardial M cells gt endocardialbull Pharmacology A common finding in cardiac disease is reduction in Ito1 density with a loss
of heterogeneity of Ito expression and associated action potential prolongation
bull Channel sources1 Transfected cell lines CHO L cell COS-7 cell HEK 293 and Xenopus Oocyte (818 = 44)2 Myocyte (56) human AM (3) rat VM (4) Canine VM 92) and rabbit VM (1)
Guinea Pig (0 very low expression of Ito)
bull Temperature ndash RT = 1215 (80)ndash near or real physiological 315 (20)
1 Pulse sequencesndash Holding -90 to -40 (mode -80 mV) ndash Activation 0 to 75 mV (mode 50 mV)ndash Duration of activation 150 ndash 500 ms (mode 300 ms)
bull Ito measurement peak current or after 70ms of activation
Pulse sequences Ito
Ito (Kir 43)
ID ChannelCurrent
Speciescells CPD Temp Vh(mV)
Prepulse(ms mV)
Activation(ms mV)
Voltage -Dependence(ms mV)
Intervalfrequency
47 hKv43hKChIP22 CHO Methadone RT -90 300 40
49 rKv43 MouseL cell Vanoxerine RT -80 300 20 15s
50 hKv43 HEK293 Alfuzosin RT -80 300 20 15s
206 Ito RatVM Zacopride 24 -40 na 500 -40 to 80 (10)
210 Ito HumanAM Cyamemazine
32-34 -50 500 60
214 Kv43 MonkeyCOS7 Pentamidine na -80 na 500 -40 to 60
215 Ito CanineVM Pioglitazone na -80 5ms -40 na 50
220 Ito RatVM Ajmaline RT -75 300 60
221 rKv42 rKv43 HEK293 RSD1235 25 -80 Brief -50 250 60 1Hz
222 Ito RatVM FK-506 22-24 -80 na 50 1000 -50 to 70 01Hz
224 hKv43 HEK293 Allitridi 22-23 -80 300 60 300 -80 to 60
245 Ito HumanAM DPO RT -80 150 40
256 Ito RabbitVM Carvedilol 34 -60 300 40 300 -50 to 60
263 Kv43 CHO (transient) Spironolactone canrenoic acid
RT -80 250 50 250 -90 to 50 10s
267 Ito ratVM Ebastine na -90 na 75 na -30 to 75
273 Ito CanineVM Thymol 37 -80 5ms- 40 400 50 400 -10 to 60
281 rKv14 Xenopus Oocyte Haloperidol 22 -80 300 0
283 Ito HumanAM Clotrimaxole 21-22 -50 300 50 300 -40 to 60 02Hz
VM= ventricular myocyte AM= Atrial myocyte
Kinetics Pulse Sequences Ito
ID channel cells kinetics Vh (mV)
1st (ms mv)
2nd (ms mV)
3rd (ms mv)
4th (ms mV)
224 hKv43 HEK293 Inactivation -80 -10 to 60
Channel availability -80 na -100 to 40 300 50
Activation conductance -80 8 na na
Recovery time from inactivation -80 30s 50 t -80 300 50 -80
Close state inactivation -90 300 50 3000 -90 t -50 300 50
283 Ito HumanAM Voltage dependence of activation and inactivation
-80 250 -100 to 30 (10) na 50
Recovery time from inactivation -80 30 -40 300 50 t -80 300 50
Extracellular buffer solution of ItoID NaCl NMDG KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-
acetate Blocker pH pH buffer
47 49 50 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 20 033 CdCl2 (01 CaL) BaCl2 (02 Ik1) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 CdCl2
214 140 4 2 1 10 74 NaOH
215 144 56 25 12 5 11 74 na
220 135 54 09 09 10 10 033 CoCl2 (2) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
222 135 4 18 1 2 11 TTX (001)Co++ (2) 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245 132 4 12 10 111 Nisoldipine (04 uM) na na
256 143 54 18 05 5 56 025 Nisoldipine (0003) or CoCl2 (2) and TTX (001) 74 NaOH
263-1 130 4 1 1 10 10 74 NaOH
263-2 140 4 1 1 10 10 TTX (003) CoCl2 (2) 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 140 5 18 1 6 6 Nifedipine (0005) 74 NaOH
283 140 5 1 5 10 033 BaCl2 (01) CdCl2 (02) 74 NaOH
NMDG N-methyl-D-glucamine 263-1 CHO cells 263-2 mouse VM
Pipette solution of ItoID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phospho creatinne
NaCl CaCl2 KOH KH2PO4 pH pH buffer
47 49 50
130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
214 110 142 10 4 10 517 72 KOH
215 45 100 1 10 na 5 72
220 25 130 1 1 5 01 5 3 725 KOH
221 130 1 5 4 01 10 72 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245 20 110 5 5 10 72
256 80 40 10 5 5 5 065 60 72
263 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
Summary of IKur (Kv1531) protocolsbull Distribution a major repolarizing current in human atria but seems to play no
role in the ventricle bull Pharmacology selective IKur blockers to restore and maintain sinus rhythm
without a risk of ventricular pro-arrhythmia
bull Channel sources1 Transfected cell lines CHO L cells HEK 293 and Xenopus Oocyte (710 =
70)2 Myocyte (30) species ndash human AM and mouse VM
bull Temperature RT = 100bull Pulse sequences
ndash Holding range -80 to -50 mV (mode -80 mV)ndash Activation 20 - 60 mV (mode 50 mV)ndash Duration of activation 150 ndash 1000 ms (mode 300 ms)
1 IKur measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKurID Channel
CurrentSpeciescells CPD Temp Vh
(mV)Prepulse Activation
(ms mV)Tail current
Voltage -Dependence(ms mV)
Interval (s)
49 hKv15 MouseL cell vanoxerine RT -80 300 20 10s
221 Kv15 HEK293 RSD1235 25 -80 200 60
224 hKv15 HEK293 Allitridi 22-23 -80 300 50 300 -40 to 60
245-1 hKv15 CHO DPO RT -80 150 40 na -30
245-2 Ikur HumanAM DPO RT -50 300 40 na -30
245-3 hKv15 XO DPO na -80 na 40
262 hKv15 CHO SSR149744C RT -70 1000 30 20s
263 hKv15 Mouse LtK- Spironolactonecanrenoic acid
RT -80 500 60 500 -40 500 -80 to 60
263 Ikur MouseVM Spironolactonecanrenoic acid
RT -80 200 40 500 50 10s
283 Ikur HumanAM clotrimazole 21-22 -50 100 40 150 50 na -30 150 -40 to 50
Ikur (Kv15)
Extracellular buffer solution of IKurID NaCl NMCG K-
gluconate KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-acetate Blocker pH pH
buffer
49 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 10 033 CdCl2 (01) BaCl2 (02) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245-1 132 4 18 12 10 111 72 na
245-2 132 4 12 10 111 nisoldipine (04)
245-3 116 4 10 725
262 140 54 2 2 15 10 74 NaOH
263 130 4 1 1 10 10 74 NaOH
281 96 2 18 1 5 74 NaOH
283 140 5 1 1 5 10 033BaCl2 (02 Ik1) CdCl2 (Ica)
74 NaOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Pipette solution of IKurID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phosphocreatinne
NaCl CaCl2 K-gluconate KH2PO4 pH pH
buffer
49 130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
221 130 1 5 4 01 10 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245-1 15 32 5 5 5 119 735
245-2 20 110 5 5 10 72
245-3 4 10 003 116 725
262 140 2 11 10 2 73 KOH
263 42 80 5 5 5 3 10 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Summary of Iks protocolsbull Distribution all cell types but less in midmyocardial myocytes
bull Pharmacology Atrial and ventricular repolarizationHeart rate dependentbull subunit KCNQ1 (channel forming) Kv71 KvLQT1bull subunit KCNE1 (accessary unit activation of KCNQ1) minK (minimum potassium
subunit)
bull Channel sources1 transfected cell lines CHO HEK 293 Cos-7 cells and Xenopus Oocyte (2136 = 58)2 Myocyte (42) species ndash Canine VM Guinea pig VM rabbit VM and mouse VM (mode
GP)
bull Temperature RT = 1629 55 physiological temperature = 45
bull Pulse sequencesndash Holding potential range = -90 to -30 (mode -80)ndash Depolarization potential 20-60 mV ndash Duration of depolarization 50 ms ndash 10s (mode 2s)ndash Repolarization potential -80 to -10 mV (mode -40 mV)ndash Duration of repolarization 05 ndash 5s (mode 2s)
bull IKs measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation (s mV)
deactivation(s mV)
Interval (s)
47 hKvLQT1hminK CHO RT -80 2 40 01 Hz
49 hKv711hKCNE1 CHO RT -80 2 20 05 -40 15s
50 hKv711hKCNE1 HEk293 RT -80 2 20 05 -40 15s
184 IKs canineepiardial cells 37 -50 2 40 na -30 na
214 Kv711minK HEK293 na -80 5 -60 to 60 5s -50 na
215 IKs caninemyocyte na -40 5 50
224 hKCNQ1hKCNE1 HEK293 22-23 -80 3 -60 to 60 (20) na -50 na
229 hCNE1-hKCNQ1 Transgenic mousemyocyte RT -75 20 -40(Na block) 2 60 2 -40 006 Hz
230 KCNQ1KCNE1 XenopusOocyte 20-22 -80 5 30 na -70
232 KCNQ1KCNE1 HEK293 36 -80 2 -80 to 110 (10) 2 -60 6s
233 KCNQ1KCNE1 CHO na -80 80 -90 3 20 1 -50 005Hz
237 KCNQ1 CHO na -80 na -80 to 60 1 -40
239 KCNQ1KCNE1 HEK293 RT -70 1 -70 to 60 na -40
241 KCNQ1 andor minK MouseCOS-7 22-25 -80 3 -120 to 60 (20) 2 -30
242 hKvLQT1hminK HEK293 19-36 -80 3 -50 to 60 (10) 5 -50
242 hKvLQT1hminK HEK293 19-36 -50 1 or 2 50 na -50 10s
244 KCNQ1 andor KCNE1 CHO na -90 3 -70 to 70 (10) na -60
245 IKs GPMyocyte 35 -50 1 50
249 KCNQ1KCNE1 HEK293 33-34 -40 1 -40 to 80 (10) na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation(s mV)
Voltage dependent(s mV)
Deactivation(s mV)
Interval (s)
251 IKs GPMyocyte 22-2535-37 -40 2 60 2 -40 to 80 (20) na -40
252 IKs GPMyocyte 35 -80 200 -50 3 60 2 -40 60s
252 hminK XenopusOocyte 35 -80 10 20 10 -40 to 60 2 -80
256 IKs Rabbitmyocyte 34 -50 3 -40 to 50 na
260 hKCNQ1hKCNE1 HEK293 RT -80 25 -70 to 100 (10) 25 -40 15s
262 IKs GPMyocyte 35 -80 1 40 5 -10
263 Kv71 and minK CHO RT -80 2 40 na -40
264 IKs GPmyocyte na -80 200 -40 05 50 na -40
267 IKs GPmyocyte na -90 na 0 na 40 01- 015 0-80 -90
273 IKs Caninemyocyte 37 -40 3 50 na -40
274 IKs GPmyocyte 36 -30 05 50 or 2 70 05 (2) -30 to 70 na -30
275 KCNQ1KCNE1 CHO 21 -80 1 40 01Hz
276 hKCNQ1hminK CHO 22 -80 6 40 na -80 005Hz
278 IKs Canine myocyte 35-37 -50 3 -50 to 50 2s -40
283 KCNQ1KCNE1 HEK293 21-22 -80 3 40 3 -80 to 40 na -40
286 IKs GPmyocyte 34-36 -50 005 30 005 ndash3 -50 to 30 na -50
287 Iks GPmyocyte 36 -80 na -40 2 60 2 -40 to 80 na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
47 49 50
137 4 18 1 10 10 74 NaOH
184 145 4 18 18 10 11 NaH2PO4 1 E4031 (5uM) Cadmium (300 uM) 74 NaOH
214 140 4 2 1 10 74 NaOH
215 144 5 56 25 5 Dextrose 11 Nifedipine (5 uM) E4031 (1 uM) 74 NaOH
224 140 54 18 1 10 10 NaH2PO4 033 73 NaOH
229 132 48 2 12 10 5 E4031 nisoldipine 74 NaOH
230 96 2 18 1 5 74 NaOH
232 140 54 18 1 5 55 74 NaOH
233 17 120 4 2 1 10 10 74 NaOH
237 130 5 18 1 10 Na-Acetate 28 74 NaOH
239 150 54 18 4 10 74 NaOH
241 135 5 25 12 5 10 74 NaOH
242 132 4 18 12 10 111 74 NaOH
244 140 4 18 12 55 11 74 NaOH
249 140 54 18 05 5 55 NaH2PO4 033 74 NaOH
250 140 54 18 1 5 10 74 NaOH
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
251 1 1 2 10 N-methly-d-glucamine 132
lanthanum chloride 005 mMcadmium chloride 02 mM 74 HCl
252 140 47 13 1 10 10 74 NaOH
256 09 5 N-methly-d-glucamine 149
Nisoldipine 3 uME4031 10 uM 74 HCl
260 140 54 18 1 5 10 NaH2PO4 03 73 NaOH
262 137 4 18 2 10 115 Pyrubic acid 5 Thiamine 01 74 NaOH
263 130 4 1 1 10 10 TTX 003 mM CoCl2 2 mM 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10 KH2PO4 12 MgSO4 12 NaHCO3 44
Nifetidine 5 uM 4-aminopyridine 3 mM 72 NaOH
274 140 1 5 10 Cd2+ 02 mM 74 NaOH
275 130 54 18 1 10 5 74 NaOH
278 136 54 1 1 5 Dextrose 10 NaH2PO4 033 E4031 5 uM 74 NaOH
283 140 5 1 1 5 10 NaH2PO4 033 74 NaOH
286 09 5 5 N-methly-d-glucamine 149
Nisoldipine 04 uME4031 5 uM 74 NaOH
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
49 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
214 110 142 10 4 4 517 72 KOH
215 45 100 1 10 na 5 72
224 20 110 1 5 5 01 10 5 72 KOH
229 110 11 5 10 55 73
230 3000
232 20 125 10 10
233 140 1 1 20 73 KOH
237 10 125 1 10 5 10 10 72 KOH
239 30 130 01 4 10 74
241 117 2 11 11 10 1 72
242 15 119 32 5 5 5 74 KOH
244 130 1 5 5 10 74 KOH
249 130 5 5 01 5 5 74 HCl
250 80 40 10 5 01 5 40 065 72 HCl
251 50 60 1 11 5 10 1 74 KOH
252 140 11 1 10 10 72 KOH
amphotericin-B
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
256 80 40 10 5 5 5 065 72
262 150 1 5 4 5 5 73 KOH
263^ 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
274 40 106 1 5 4 5 72 KOH
276 20 110 1 10 5 10 10 1 72 KOH
278 20 110 1 10 5 01 10 5 72 KOH
283 20 110 1 5 5 01 1 5 72 KOH
286 50 1 5 5 01 5 2 10 72 KOH
^phosphatidylinositol 45-diphosphate (5 mgml 1) was added to the internal solution to avoid the rundown of the current 70 L-aspartic acid 70 KOH
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Pulse sequences Ito
Ito (Kir 43)
ID ChannelCurrent
Speciescells CPD Temp Vh(mV)
Prepulse(ms mV)
Activation(ms mV)
Voltage -Dependence(ms mV)
Intervalfrequency
47 hKv43hKChIP22 CHO Methadone RT -90 300 40
49 rKv43 MouseL cell Vanoxerine RT -80 300 20 15s
50 hKv43 HEK293 Alfuzosin RT -80 300 20 15s
206 Ito RatVM Zacopride 24 -40 na 500 -40 to 80 (10)
210 Ito HumanAM Cyamemazine
32-34 -50 500 60
214 Kv43 MonkeyCOS7 Pentamidine na -80 na 500 -40 to 60
215 Ito CanineVM Pioglitazone na -80 5ms -40 na 50
220 Ito RatVM Ajmaline RT -75 300 60
221 rKv42 rKv43 HEK293 RSD1235 25 -80 Brief -50 250 60 1Hz
222 Ito RatVM FK-506 22-24 -80 na 50 1000 -50 to 70 01Hz
224 hKv43 HEK293 Allitridi 22-23 -80 300 60 300 -80 to 60
245 Ito HumanAM DPO RT -80 150 40
256 Ito RabbitVM Carvedilol 34 -60 300 40 300 -50 to 60
263 Kv43 CHO (transient) Spironolactone canrenoic acid
RT -80 250 50 250 -90 to 50 10s
267 Ito ratVM Ebastine na -90 na 75 na -30 to 75
273 Ito CanineVM Thymol 37 -80 5ms- 40 400 50 400 -10 to 60
281 rKv14 Xenopus Oocyte Haloperidol 22 -80 300 0
283 Ito HumanAM Clotrimaxole 21-22 -50 300 50 300 -40 to 60 02Hz
VM= ventricular myocyte AM= Atrial myocyte
Kinetics Pulse Sequences Ito
ID channel cells kinetics Vh (mV)
1st (ms mv)
2nd (ms mV)
3rd (ms mv)
4th (ms mV)
224 hKv43 HEK293 Inactivation -80 -10 to 60
Channel availability -80 na -100 to 40 300 50
Activation conductance -80 8 na na
Recovery time from inactivation -80 30s 50 t -80 300 50 -80
Close state inactivation -90 300 50 3000 -90 t -50 300 50
283 Ito HumanAM Voltage dependence of activation and inactivation
-80 250 -100 to 30 (10) na 50
Recovery time from inactivation -80 30 -40 300 50 t -80 300 50
Extracellular buffer solution of ItoID NaCl NMDG KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-
acetate Blocker pH pH buffer
47 49 50 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 20 033 CdCl2 (01 CaL) BaCl2 (02 Ik1) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 CdCl2
214 140 4 2 1 10 74 NaOH
215 144 56 25 12 5 11 74 na
220 135 54 09 09 10 10 033 CoCl2 (2) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
222 135 4 18 1 2 11 TTX (001)Co++ (2) 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245 132 4 12 10 111 Nisoldipine (04 uM) na na
256 143 54 18 05 5 56 025 Nisoldipine (0003) or CoCl2 (2) and TTX (001) 74 NaOH
263-1 130 4 1 1 10 10 74 NaOH
263-2 140 4 1 1 10 10 TTX (003) CoCl2 (2) 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 140 5 18 1 6 6 Nifedipine (0005) 74 NaOH
283 140 5 1 5 10 033 BaCl2 (01) CdCl2 (02) 74 NaOH
NMDG N-methyl-D-glucamine 263-1 CHO cells 263-2 mouse VM
Pipette solution of ItoID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phospho creatinne
NaCl CaCl2 KOH KH2PO4 pH pH buffer
47 49 50
130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
214 110 142 10 4 10 517 72 KOH
215 45 100 1 10 na 5 72
220 25 130 1 1 5 01 5 3 725 KOH
221 130 1 5 4 01 10 72 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245 20 110 5 5 10 72
256 80 40 10 5 5 5 065 60 72
263 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
Summary of IKur (Kv1531) protocolsbull Distribution a major repolarizing current in human atria but seems to play no
role in the ventricle bull Pharmacology selective IKur blockers to restore and maintain sinus rhythm
without a risk of ventricular pro-arrhythmia
bull Channel sources1 Transfected cell lines CHO L cells HEK 293 and Xenopus Oocyte (710 =
70)2 Myocyte (30) species ndash human AM and mouse VM
bull Temperature RT = 100bull Pulse sequences
ndash Holding range -80 to -50 mV (mode -80 mV)ndash Activation 20 - 60 mV (mode 50 mV)ndash Duration of activation 150 ndash 1000 ms (mode 300 ms)
1 IKur measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKurID Channel
CurrentSpeciescells CPD Temp Vh
(mV)Prepulse Activation
(ms mV)Tail current
Voltage -Dependence(ms mV)
Interval (s)
49 hKv15 MouseL cell vanoxerine RT -80 300 20 10s
221 Kv15 HEK293 RSD1235 25 -80 200 60
224 hKv15 HEK293 Allitridi 22-23 -80 300 50 300 -40 to 60
245-1 hKv15 CHO DPO RT -80 150 40 na -30
245-2 Ikur HumanAM DPO RT -50 300 40 na -30
245-3 hKv15 XO DPO na -80 na 40
262 hKv15 CHO SSR149744C RT -70 1000 30 20s
263 hKv15 Mouse LtK- Spironolactonecanrenoic acid
RT -80 500 60 500 -40 500 -80 to 60
263 Ikur MouseVM Spironolactonecanrenoic acid
RT -80 200 40 500 50 10s
283 Ikur HumanAM clotrimazole 21-22 -50 100 40 150 50 na -30 150 -40 to 50
Ikur (Kv15)
Extracellular buffer solution of IKurID NaCl NMCG K-
gluconate KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-acetate Blocker pH pH
buffer
49 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 10 033 CdCl2 (01) BaCl2 (02) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245-1 132 4 18 12 10 111 72 na
245-2 132 4 12 10 111 nisoldipine (04)
245-3 116 4 10 725
262 140 54 2 2 15 10 74 NaOH
263 130 4 1 1 10 10 74 NaOH
281 96 2 18 1 5 74 NaOH
283 140 5 1 1 5 10 033BaCl2 (02 Ik1) CdCl2 (Ica)
74 NaOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Pipette solution of IKurID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phosphocreatinne
NaCl CaCl2 K-gluconate KH2PO4 pH pH
buffer
49 130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
221 130 1 5 4 01 10 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245-1 15 32 5 5 5 119 735
245-2 20 110 5 5 10 72
245-3 4 10 003 116 725
262 140 2 11 10 2 73 KOH
263 42 80 5 5 5 3 10 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Summary of Iks protocolsbull Distribution all cell types but less in midmyocardial myocytes
bull Pharmacology Atrial and ventricular repolarizationHeart rate dependentbull subunit KCNQ1 (channel forming) Kv71 KvLQT1bull subunit KCNE1 (accessary unit activation of KCNQ1) minK (minimum potassium
subunit)
bull Channel sources1 transfected cell lines CHO HEK 293 Cos-7 cells and Xenopus Oocyte (2136 = 58)2 Myocyte (42) species ndash Canine VM Guinea pig VM rabbit VM and mouse VM (mode
GP)
bull Temperature RT = 1629 55 physiological temperature = 45
bull Pulse sequencesndash Holding potential range = -90 to -30 (mode -80)ndash Depolarization potential 20-60 mV ndash Duration of depolarization 50 ms ndash 10s (mode 2s)ndash Repolarization potential -80 to -10 mV (mode -40 mV)ndash Duration of repolarization 05 ndash 5s (mode 2s)
bull IKs measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation (s mV)
deactivation(s mV)
Interval (s)
47 hKvLQT1hminK CHO RT -80 2 40 01 Hz
49 hKv711hKCNE1 CHO RT -80 2 20 05 -40 15s
50 hKv711hKCNE1 HEk293 RT -80 2 20 05 -40 15s
184 IKs canineepiardial cells 37 -50 2 40 na -30 na
214 Kv711minK HEK293 na -80 5 -60 to 60 5s -50 na
215 IKs caninemyocyte na -40 5 50
224 hKCNQ1hKCNE1 HEK293 22-23 -80 3 -60 to 60 (20) na -50 na
229 hCNE1-hKCNQ1 Transgenic mousemyocyte RT -75 20 -40(Na block) 2 60 2 -40 006 Hz
230 KCNQ1KCNE1 XenopusOocyte 20-22 -80 5 30 na -70
232 KCNQ1KCNE1 HEK293 36 -80 2 -80 to 110 (10) 2 -60 6s
233 KCNQ1KCNE1 CHO na -80 80 -90 3 20 1 -50 005Hz
237 KCNQ1 CHO na -80 na -80 to 60 1 -40
239 KCNQ1KCNE1 HEK293 RT -70 1 -70 to 60 na -40
241 KCNQ1 andor minK MouseCOS-7 22-25 -80 3 -120 to 60 (20) 2 -30
242 hKvLQT1hminK HEK293 19-36 -80 3 -50 to 60 (10) 5 -50
242 hKvLQT1hminK HEK293 19-36 -50 1 or 2 50 na -50 10s
244 KCNQ1 andor KCNE1 CHO na -90 3 -70 to 70 (10) na -60
245 IKs GPMyocyte 35 -50 1 50
249 KCNQ1KCNE1 HEK293 33-34 -40 1 -40 to 80 (10) na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation(s mV)
Voltage dependent(s mV)
Deactivation(s mV)
Interval (s)
251 IKs GPMyocyte 22-2535-37 -40 2 60 2 -40 to 80 (20) na -40
252 IKs GPMyocyte 35 -80 200 -50 3 60 2 -40 60s
252 hminK XenopusOocyte 35 -80 10 20 10 -40 to 60 2 -80
256 IKs Rabbitmyocyte 34 -50 3 -40 to 50 na
260 hKCNQ1hKCNE1 HEK293 RT -80 25 -70 to 100 (10) 25 -40 15s
262 IKs GPMyocyte 35 -80 1 40 5 -10
263 Kv71 and minK CHO RT -80 2 40 na -40
264 IKs GPmyocyte na -80 200 -40 05 50 na -40
267 IKs GPmyocyte na -90 na 0 na 40 01- 015 0-80 -90
273 IKs Caninemyocyte 37 -40 3 50 na -40
274 IKs GPmyocyte 36 -30 05 50 or 2 70 05 (2) -30 to 70 na -30
275 KCNQ1KCNE1 CHO 21 -80 1 40 01Hz
276 hKCNQ1hminK CHO 22 -80 6 40 na -80 005Hz
278 IKs Canine myocyte 35-37 -50 3 -50 to 50 2s -40
283 KCNQ1KCNE1 HEK293 21-22 -80 3 40 3 -80 to 40 na -40
286 IKs GPmyocyte 34-36 -50 005 30 005 ndash3 -50 to 30 na -50
287 Iks GPmyocyte 36 -80 na -40 2 60 2 -40 to 80 na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
47 49 50
137 4 18 1 10 10 74 NaOH
184 145 4 18 18 10 11 NaH2PO4 1 E4031 (5uM) Cadmium (300 uM) 74 NaOH
214 140 4 2 1 10 74 NaOH
215 144 5 56 25 5 Dextrose 11 Nifedipine (5 uM) E4031 (1 uM) 74 NaOH
224 140 54 18 1 10 10 NaH2PO4 033 73 NaOH
229 132 48 2 12 10 5 E4031 nisoldipine 74 NaOH
230 96 2 18 1 5 74 NaOH
232 140 54 18 1 5 55 74 NaOH
233 17 120 4 2 1 10 10 74 NaOH
237 130 5 18 1 10 Na-Acetate 28 74 NaOH
239 150 54 18 4 10 74 NaOH
241 135 5 25 12 5 10 74 NaOH
242 132 4 18 12 10 111 74 NaOH
244 140 4 18 12 55 11 74 NaOH
249 140 54 18 05 5 55 NaH2PO4 033 74 NaOH
250 140 54 18 1 5 10 74 NaOH
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
251 1 1 2 10 N-methly-d-glucamine 132
lanthanum chloride 005 mMcadmium chloride 02 mM 74 HCl
252 140 47 13 1 10 10 74 NaOH
256 09 5 N-methly-d-glucamine 149
Nisoldipine 3 uME4031 10 uM 74 HCl
260 140 54 18 1 5 10 NaH2PO4 03 73 NaOH
262 137 4 18 2 10 115 Pyrubic acid 5 Thiamine 01 74 NaOH
263 130 4 1 1 10 10 TTX 003 mM CoCl2 2 mM 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10 KH2PO4 12 MgSO4 12 NaHCO3 44
Nifetidine 5 uM 4-aminopyridine 3 mM 72 NaOH
274 140 1 5 10 Cd2+ 02 mM 74 NaOH
275 130 54 18 1 10 5 74 NaOH
278 136 54 1 1 5 Dextrose 10 NaH2PO4 033 E4031 5 uM 74 NaOH
283 140 5 1 1 5 10 NaH2PO4 033 74 NaOH
286 09 5 5 N-methly-d-glucamine 149
Nisoldipine 04 uME4031 5 uM 74 NaOH
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
49 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
214 110 142 10 4 4 517 72 KOH
215 45 100 1 10 na 5 72
224 20 110 1 5 5 01 10 5 72 KOH
229 110 11 5 10 55 73
230 3000
232 20 125 10 10
233 140 1 1 20 73 KOH
237 10 125 1 10 5 10 10 72 KOH
239 30 130 01 4 10 74
241 117 2 11 11 10 1 72
242 15 119 32 5 5 5 74 KOH
244 130 1 5 5 10 74 KOH
249 130 5 5 01 5 5 74 HCl
250 80 40 10 5 01 5 40 065 72 HCl
251 50 60 1 11 5 10 1 74 KOH
252 140 11 1 10 10 72 KOH
amphotericin-B
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
256 80 40 10 5 5 5 065 72
262 150 1 5 4 5 5 73 KOH
263^ 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
274 40 106 1 5 4 5 72 KOH
276 20 110 1 10 5 10 10 1 72 KOH
278 20 110 1 10 5 01 10 5 72 KOH
283 20 110 1 5 5 01 1 5 72 KOH
286 50 1 5 5 01 5 2 10 72 KOH
^phosphatidylinositol 45-diphosphate (5 mgml 1) was added to the internal solution to avoid the rundown of the current 70 L-aspartic acid 70 KOH
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Kinetics Pulse Sequences Ito
ID channel cells kinetics Vh (mV)
1st (ms mv)
2nd (ms mV)
3rd (ms mv)
4th (ms mV)
224 hKv43 HEK293 Inactivation -80 -10 to 60
Channel availability -80 na -100 to 40 300 50
Activation conductance -80 8 na na
Recovery time from inactivation -80 30s 50 t -80 300 50 -80
Close state inactivation -90 300 50 3000 -90 t -50 300 50
283 Ito HumanAM Voltage dependence of activation and inactivation
-80 250 -100 to 30 (10) na 50
Recovery time from inactivation -80 30 -40 300 50 t -80 300 50
Extracellular buffer solution of ItoID NaCl NMDG KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-
acetate Blocker pH pH buffer
47 49 50 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 20 033 CdCl2 (01 CaL) BaCl2 (02 Ik1) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 CdCl2
214 140 4 2 1 10 74 NaOH
215 144 56 25 12 5 11 74 na
220 135 54 09 09 10 10 033 CoCl2 (2) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
222 135 4 18 1 2 11 TTX (001)Co++ (2) 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245 132 4 12 10 111 Nisoldipine (04 uM) na na
256 143 54 18 05 5 56 025 Nisoldipine (0003) or CoCl2 (2) and TTX (001) 74 NaOH
263-1 130 4 1 1 10 10 74 NaOH
263-2 140 4 1 1 10 10 TTX (003) CoCl2 (2) 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 140 5 18 1 6 6 Nifedipine (0005) 74 NaOH
283 140 5 1 5 10 033 BaCl2 (01) CdCl2 (02) 74 NaOH
NMDG N-methyl-D-glucamine 263-1 CHO cells 263-2 mouse VM
Pipette solution of ItoID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phospho creatinne
NaCl CaCl2 KOH KH2PO4 pH pH buffer
47 49 50
130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
214 110 142 10 4 10 517 72 KOH
215 45 100 1 10 na 5 72
220 25 130 1 1 5 01 5 3 725 KOH
221 130 1 5 4 01 10 72 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245 20 110 5 5 10 72
256 80 40 10 5 5 5 065 60 72
263 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
Summary of IKur (Kv1531) protocolsbull Distribution a major repolarizing current in human atria but seems to play no
role in the ventricle bull Pharmacology selective IKur blockers to restore and maintain sinus rhythm
without a risk of ventricular pro-arrhythmia
bull Channel sources1 Transfected cell lines CHO L cells HEK 293 and Xenopus Oocyte (710 =
70)2 Myocyte (30) species ndash human AM and mouse VM
bull Temperature RT = 100bull Pulse sequences
ndash Holding range -80 to -50 mV (mode -80 mV)ndash Activation 20 - 60 mV (mode 50 mV)ndash Duration of activation 150 ndash 1000 ms (mode 300 ms)
1 IKur measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKurID Channel
CurrentSpeciescells CPD Temp Vh
(mV)Prepulse Activation
(ms mV)Tail current
Voltage -Dependence(ms mV)
Interval (s)
49 hKv15 MouseL cell vanoxerine RT -80 300 20 10s
221 Kv15 HEK293 RSD1235 25 -80 200 60
224 hKv15 HEK293 Allitridi 22-23 -80 300 50 300 -40 to 60
245-1 hKv15 CHO DPO RT -80 150 40 na -30
245-2 Ikur HumanAM DPO RT -50 300 40 na -30
245-3 hKv15 XO DPO na -80 na 40
262 hKv15 CHO SSR149744C RT -70 1000 30 20s
263 hKv15 Mouse LtK- Spironolactonecanrenoic acid
RT -80 500 60 500 -40 500 -80 to 60
263 Ikur MouseVM Spironolactonecanrenoic acid
RT -80 200 40 500 50 10s
283 Ikur HumanAM clotrimazole 21-22 -50 100 40 150 50 na -30 150 -40 to 50
Ikur (Kv15)
Extracellular buffer solution of IKurID NaCl NMCG K-
gluconate KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-acetate Blocker pH pH
buffer
49 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 10 033 CdCl2 (01) BaCl2 (02) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245-1 132 4 18 12 10 111 72 na
245-2 132 4 12 10 111 nisoldipine (04)
245-3 116 4 10 725
262 140 54 2 2 15 10 74 NaOH
263 130 4 1 1 10 10 74 NaOH
281 96 2 18 1 5 74 NaOH
283 140 5 1 1 5 10 033BaCl2 (02 Ik1) CdCl2 (Ica)
74 NaOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Pipette solution of IKurID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phosphocreatinne
NaCl CaCl2 K-gluconate KH2PO4 pH pH
buffer
49 130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
221 130 1 5 4 01 10 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245-1 15 32 5 5 5 119 735
245-2 20 110 5 5 10 72
245-3 4 10 003 116 725
262 140 2 11 10 2 73 KOH
263 42 80 5 5 5 3 10 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Summary of Iks protocolsbull Distribution all cell types but less in midmyocardial myocytes
bull Pharmacology Atrial and ventricular repolarizationHeart rate dependentbull subunit KCNQ1 (channel forming) Kv71 KvLQT1bull subunit KCNE1 (accessary unit activation of KCNQ1) minK (minimum potassium
subunit)
bull Channel sources1 transfected cell lines CHO HEK 293 Cos-7 cells and Xenopus Oocyte (2136 = 58)2 Myocyte (42) species ndash Canine VM Guinea pig VM rabbit VM and mouse VM (mode
GP)
bull Temperature RT = 1629 55 physiological temperature = 45
bull Pulse sequencesndash Holding potential range = -90 to -30 (mode -80)ndash Depolarization potential 20-60 mV ndash Duration of depolarization 50 ms ndash 10s (mode 2s)ndash Repolarization potential -80 to -10 mV (mode -40 mV)ndash Duration of repolarization 05 ndash 5s (mode 2s)
bull IKs measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation (s mV)
deactivation(s mV)
Interval (s)
47 hKvLQT1hminK CHO RT -80 2 40 01 Hz
49 hKv711hKCNE1 CHO RT -80 2 20 05 -40 15s
50 hKv711hKCNE1 HEk293 RT -80 2 20 05 -40 15s
184 IKs canineepiardial cells 37 -50 2 40 na -30 na
214 Kv711minK HEK293 na -80 5 -60 to 60 5s -50 na
215 IKs caninemyocyte na -40 5 50
224 hKCNQ1hKCNE1 HEK293 22-23 -80 3 -60 to 60 (20) na -50 na
229 hCNE1-hKCNQ1 Transgenic mousemyocyte RT -75 20 -40(Na block) 2 60 2 -40 006 Hz
230 KCNQ1KCNE1 XenopusOocyte 20-22 -80 5 30 na -70
232 KCNQ1KCNE1 HEK293 36 -80 2 -80 to 110 (10) 2 -60 6s
233 KCNQ1KCNE1 CHO na -80 80 -90 3 20 1 -50 005Hz
237 KCNQ1 CHO na -80 na -80 to 60 1 -40
239 KCNQ1KCNE1 HEK293 RT -70 1 -70 to 60 na -40
241 KCNQ1 andor minK MouseCOS-7 22-25 -80 3 -120 to 60 (20) 2 -30
242 hKvLQT1hminK HEK293 19-36 -80 3 -50 to 60 (10) 5 -50
242 hKvLQT1hminK HEK293 19-36 -50 1 or 2 50 na -50 10s
244 KCNQ1 andor KCNE1 CHO na -90 3 -70 to 70 (10) na -60
245 IKs GPMyocyte 35 -50 1 50
249 KCNQ1KCNE1 HEK293 33-34 -40 1 -40 to 80 (10) na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation(s mV)
Voltage dependent(s mV)
Deactivation(s mV)
Interval (s)
251 IKs GPMyocyte 22-2535-37 -40 2 60 2 -40 to 80 (20) na -40
252 IKs GPMyocyte 35 -80 200 -50 3 60 2 -40 60s
252 hminK XenopusOocyte 35 -80 10 20 10 -40 to 60 2 -80
256 IKs Rabbitmyocyte 34 -50 3 -40 to 50 na
260 hKCNQ1hKCNE1 HEK293 RT -80 25 -70 to 100 (10) 25 -40 15s
262 IKs GPMyocyte 35 -80 1 40 5 -10
263 Kv71 and minK CHO RT -80 2 40 na -40
264 IKs GPmyocyte na -80 200 -40 05 50 na -40
267 IKs GPmyocyte na -90 na 0 na 40 01- 015 0-80 -90
273 IKs Caninemyocyte 37 -40 3 50 na -40
274 IKs GPmyocyte 36 -30 05 50 or 2 70 05 (2) -30 to 70 na -30
275 KCNQ1KCNE1 CHO 21 -80 1 40 01Hz
276 hKCNQ1hminK CHO 22 -80 6 40 na -80 005Hz
278 IKs Canine myocyte 35-37 -50 3 -50 to 50 2s -40
283 KCNQ1KCNE1 HEK293 21-22 -80 3 40 3 -80 to 40 na -40
286 IKs GPmyocyte 34-36 -50 005 30 005 ndash3 -50 to 30 na -50
287 Iks GPmyocyte 36 -80 na -40 2 60 2 -40 to 80 na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
47 49 50
137 4 18 1 10 10 74 NaOH
184 145 4 18 18 10 11 NaH2PO4 1 E4031 (5uM) Cadmium (300 uM) 74 NaOH
214 140 4 2 1 10 74 NaOH
215 144 5 56 25 5 Dextrose 11 Nifedipine (5 uM) E4031 (1 uM) 74 NaOH
224 140 54 18 1 10 10 NaH2PO4 033 73 NaOH
229 132 48 2 12 10 5 E4031 nisoldipine 74 NaOH
230 96 2 18 1 5 74 NaOH
232 140 54 18 1 5 55 74 NaOH
233 17 120 4 2 1 10 10 74 NaOH
237 130 5 18 1 10 Na-Acetate 28 74 NaOH
239 150 54 18 4 10 74 NaOH
241 135 5 25 12 5 10 74 NaOH
242 132 4 18 12 10 111 74 NaOH
244 140 4 18 12 55 11 74 NaOH
249 140 54 18 05 5 55 NaH2PO4 033 74 NaOH
250 140 54 18 1 5 10 74 NaOH
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
251 1 1 2 10 N-methly-d-glucamine 132
lanthanum chloride 005 mMcadmium chloride 02 mM 74 HCl
252 140 47 13 1 10 10 74 NaOH
256 09 5 N-methly-d-glucamine 149
Nisoldipine 3 uME4031 10 uM 74 HCl
260 140 54 18 1 5 10 NaH2PO4 03 73 NaOH
262 137 4 18 2 10 115 Pyrubic acid 5 Thiamine 01 74 NaOH
263 130 4 1 1 10 10 TTX 003 mM CoCl2 2 mM 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10 KH2PO4 12 MgSO4 12 NaHCO3 44
Nifetidine 5 uM 4-aminopyridine 3 mM 72 NaOH
274 140 1 5 10 Cd2+ 02 mM 74 NaOH
275 130 54 18 1 10 5 74 NaOH
278 136 54 1 1 5 Dextrose 10 NaH2PO4 033 E4031 5 uM 74 NaOH
283 140 5 1 1 5 10 NaH2PO4 033 74 NaOH
286 09 5 5 N-methly-d-glucamine 149
Nisoldipine 04 uME4031 5 uM 74 NaOH
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
49 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
214 110 142 10 4 4 517 72 KOH
215 45 100 1 10 na 5 72
224 20 110 1 5 5 01 10 5 72 KOH
229 110 11 5 10 55 73
230 3000
232 20 125 10 10
233 140 1 1 20 73 KOH
237 10 125 1 10 5 10 10 72 KOH
239 30 130 01 4 10 74
241 117 2 11 11 10 1 72
242 15 119 32 5 5 5 74 KOH
244 130 1 5 5 10 74 KOH
249 130 5 5 01 5 5 74 HCl
250 80 40 10 5 01 5 40 065 72 HCl
251 50 60 1 11 5 10 1 74 KOH
252 140 11 1 10 10 72 KOH
amphotericin-B
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
256 80 40 10 5 5 5 065 72
262 150 1 5 4 5 5 73 KOH
263^ 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
274 40 106 1 5 4 5 72 KOH
276 20 110 1 10 5 10 10 1 72 KOH
278 20 110 1 10 5 01 10 5 72 KOH
283 20 110 1 5 5 01 1 5 72 KOH
286 50 1 5 5 01 5 2 10 72 KOH
^phosphatidylinositol 45-diphosphate (5 mgml 1) was added to the internal solution to avoid the rundown of the current 70 L-aspartic acid 70 KOH
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Extracellular buffer solution of ItoID NaCl NMDG KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-
acetate Blocker pH pH buffer
47 49 50 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 20 033 CdCl2 (01 CaL) BaCl2 (02 Ik1) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 CdCl2
214 140 4 2 1 10 74 NaOH
215 144 56 25 12 5 11 74 na
220 135 54 09 09 10 10 033 CoCl2 (2) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
222 135 4 18 1 2 11 TTX (001)Co++ (2) 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245 132 4 12 10 111 Nisoldipine (04 uM) na na
256 143 54 18 05 5 56 025 Nisoldipine (0003) or CoCl2 (2) and TTX (001) 74 NaOH
263-1 130 4 1 1 10 10 74 NaOH
263-2 140 4 1 1 10 10 TTX (003) CoCl2 (2) 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 140 5 18 1 6 6 Nifedipine (0005) 74 NaOH
283 140 5 1 5 10 033 BaCl2 (01) CdCl2 (02) 74 NaOH
NMDG N-methyl-D-glucamine 263-1 CHO cells 263-2 mouse VM
Pipette solution of ItoID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phospho creatinne
NaCl CaCl2 KOH KH2PO4 pH pH buffer
47 49 50
130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
214 110 142 10 4 10 517 72 KOH
215 45 100 1 10 na 5 72
220 25 130 1 1 5 01 5 3 725 KOH
221 130 1 5 4 01 10 72 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245 20 110 5 5 10 72
256 80 40 10 5 5 5 065 60 72
263 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
Summary of IKur (Kv1531) protocolsbull Distribution a major repolarizing current in human atria but seems to play no
role in the ventricle bull Pharmacology selective IKur blockers to restore and maintain sinus rhythm
without a risk of ventricular pro-arrhythmia
bull Channel sources1 Transfected cell lines CHO L cells HEK 293 and Xenopus Oocyte (710 =
70)2 Myocyte (30) species ndash human AM and mouse VM
bull Temperature RT = 100bull Pulse sequences
ndash Holding range -80 to -50 mV (mode -80 mV)ndash Activation 20 - 60 mV (mode 50 mV)ndash Duration of activation 150 ndash 1000 ms (mode 300 ms)
1 IKur measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKurID Channel
CurrentSpeciescells CPD Temp Vh
(mV)Prepulse Activation
(ms mV)Tail current
Voltage -Dependence(ms mV)
Interval (s)
49 hKv15 MouseL cell vanoxerine RT -80 300 20 10s
221 Kv15 HEK293 RSD1235 25 -80 200 60
224 hKv15 HEK293 Allitridi 22-23 -80 300 50 300 -40 to 60
245-1 hKv15 CHO DPO RT -80 150 40 na -30
245-2 Ikur HumanAM DPO RT -50 300 40 na -30
245-3 hKv15 XO DPO na -80 na 40
262 hKv15 CHO SSR149744C RT -70 1000 30 20s
263 hKv15 Mouse LtK- Spironolactonecanrenoic acid
RT -80 500 60 500 -40 500 -80 to 60
263 Ikur MouseVM Spironolactonecanrenoic acid
RT -80 200 40 500 50 10s
283 Ikur HumanAM clotrimazole 21-22 -50 100 40 150 50 na -30 150 -40 to 50
Ikur (Kv15)
Extracellular buffer solution of IKurID NaCl NMCG K-
gluconate KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-acetate Blocker pH pH
buffer
49 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 10 033 CdCl2 (01) BaCl2 (02) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245-1 132 4 18 12 10 111 72 na
245-2 132 4 12 10 111 nisoldipine (04)
245-3 116 4 10 725
262 140 54 2 2 15 10 74 NaOH
263 130 4 1 1 10 10 74 NaOH
281 96 2 18 1 5 74 NaOH
283 140 5 1 1 5 10 033BaCl2 (02 Ik1) CdCl2 (Ica)
74 NaOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Pipette solution of IKurID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phosphocreatinne
NaCl CaCl2 K-gluconate KH2PO4 pH pH
buffer
49 130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
221 130 1 5 4 01 10 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245-1 15 32 5 5 5 119 735
245-2 20 110 5 5 10 72
245-3 4 10 003 116 725
262 140 2 11 10 2 73 KOH
263 42 80 5 5 5 3 10 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Summary of Iks protocolsbull Distribution all cell types but less in midmyocardial myocytes
bull Pharmacology Atrial and ventricular repolarizationHeart rate dependentbull subunit KCNQ1 (channel forming) Kv71 KvLQT1bull subunit KCNE1 (accessary unit activation of KCNQ1) minK (minimum potassium
subunit)
bull Channel sources1 transfected cell lines CHO HEK 293 Cos-7 cells and Xenopus Oocyte (2136 = 58)2 Myocyte (42) species ndash Canine VM Guinea pig VM rabbit VM and mouse VM (mode
GP)
bull Temperature RT = 1629 55 physiological temperature = 45
bull Pulse sequencesndash Holding potential range = -90 to -30 (mode -80)ndash Depolarization potential 20-60 mV ndash Duration of depolarization 50 ms ndash 10s (mode 2s)ndash Repolarization potential -80 to -10 mV (mode -40 mV)ndash Duration of repolarization 05 ndash 5s (mode 2s)
bull IKs measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation (s mV)
deactivation(s mV)
Interval (s)
47 hKvLQT1hminK CHO RT -80 2 40 01 Hz
49 hKv711hKCNE1 CHO RT -80 2 20 05 -40 15s
50 hKv711hKCNE1 HEk293 RT -80 2 20 05 -40 15s
184 IKs canineepiardial cells 37 -50 2 40 na -30 na
214 Kv711minK HEK293 na -80 5 -60 to 60 5s -50 na
215 IKs caninemyocyte na -40 5 50
224 hKCNQ1hKCNE1 HEK293 22-23 -80 3 -60 to 60 (20) na -50 na
229 hCNE1-hKCNQ1 Transgenic mousemyocyte RT -75 20 -40(Na block) 2 60 2 -40 006 Hz
230 KCNQ1KCNE1 XenopusOocyte 20-22 -80 5 30 na -70
232 KCNQ1KCNE1 HEK293 36 -80 2 -80 to 110 (10) 2 -60 6s
233 KCNQ1KCNE1 CHO na -80 80 -90 3 20 1 -50 005Hz
237 KCNQ1 CHO na -80 na -80 to 60 1 -40
239 KCNQ1KCNE1 HEK293 RT -70 1 -70 to 60 na -40
241 KCNQ1 andor minK MouseCOS-7 22-25 -80 3 -120 to 60 (20) 2 -30
242 hKvLQT1hminK HEK293 19-36 -80 3 -50 to 60 (10) 5 -50
242 hKvLQT1hminK HEK293 19-36 -50 1 or 2 50 na -50 10s
244 KCNQ1 andor KCNE1 CHO na -90 3 -70 to 70 (10) na -60
245 IKs GPMyocyte 35 -50 1 50
249 KCNQ1KCNE1 HEK293 33-34 -40 1 -40 to 80 (10) na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation(s mV)
Voltage dependent(s mV)
Deactivation(s mV)
Interval (s)
251 IKs GPMyocyte 22-2535-37 -40 2 60 2 -40 to 80 (20) na -40
252 IKs GPMyocyte 35 -80 200 -50 3 60 2 -40 60s
252 hminK XenopusOocyte 35 -80 10 20 10 -40 to 60 2 -80
256 IKs Rabbitmyocyte 34 -50 3 -40 to 50 na
260 hKCNQ1hKCNE1 HEK293 RT -80 25 -70 to 100 (10) 25 -40 15s
262 IKs GPMyocyte 35 -80 1 40 5 -10
263 Kv71 and minK CHO RT -80 2 40 na -40
264 IKs GPmyocyte na -80 200 -40 05 50 na -40
267 IKs GPmyocyte na -90 na 0 na 40 01- 015 0-80 -90
273 IKs Caninemyocyte 37 -40 3 50 na -40
274 IKs GPmyocyte 36 -30 05 50 or 2 70 05 (2) -30 to 70 na -30
275 KCNQ1KCNE1 CHO 21 -80 1 40 01Hz
276 hKCNQ1hminK CHO 22 -80 6 40 na -80 005Hz
278 IKs Canine myocyte 35-37 -50 3 -50 to 50 2s -40
283 KCNQ1KCNE1 HEK293 21-22 -80 3 40 3 -80 to 40 na -40
286 IKs GPmyocyte 34-36 -50 005 30 005 ndash3 -50 to 30 na -50
287 Iks GPmyocyte 36 -80 na -40 2 60 2 -40 to 80 na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
47 49 50
137 4 18 1 10 10 74 NaOH
184 145 4 18 18 10 11 NaH2PO4 1 E4031 (5uM) Cadmium (300 uM) 74 NaOH
214 140 4 2 1 10 74 NaOH
215 144 5 56 25 5 Dextrose 11 Nifedipine (5 uM) E4031 (1 uM) 74 NaOH
224 140 54 18 1 10 10 NaH2PO4 033 73 NaOH
229 132 48 2 12 10 5 E4031 nisoldipine 74 NaOH
230 96 2 18 1 5 74 NaOH
232 140 54 18 1 5 55 74 NaOH
233 17 120 4 2 1 10 10 74 NaOH
237 130 5 18 1 10 Na-Acetate 28 74 NaOH
239 150 54 18 4 10 74 NaOH
241 135 5 25 12 5 10 74 NaOH
242 132 4 18 12 10 111 74 NaOH
244 140 4 18 12 55 11 74 NaOH
249 140 54 18 05 5 55 NaH2PO4 033 74 NaOH
250 140 54 18 1 5 10 74 NaOH
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
251 1 1 2 10 N-methly-d-glucamine 132
lanthanum chloride 005 mMcadmium chloride 02 mM 74 HCl
252 140 47 13 1 10 10 74 NaOH
256 09 5 N-methly-d-glucamine 149
Nisoldipine 3 uME4031 10 uM 74 HCl
260 140 54 18 1 5 10 NaH2PO4 03 73 NaOH
262 137 4 18 2 10 115 Pyrubic acid 5 Thiamine 01 74 NaOH
263 130 4 1 1 10 10 TTX 003 mM CoCl2 2 mM 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10 KH2PO4 12 MgSO4 12 NaHCO3 44
Nifetidine 5 uM 4-aminopyridine 3 mM 72 NaOH
274 140 1 5 10 Cd2+ 02 mM 74 NaOH
275 130 54 18 1 10 5 74 NaOH
278 136 54 1 1 5 Dextrose 10 NaH2PO4 033 E4031 5 uM 74 NaOH
283 140 5 1 1 5 10 NaH2PO4 033 74 NaOH
286 09 5 5 N-methly-d-glucamine 149
Nisoldipine 04 uME4031 5 uM 74 NaOH
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
49 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
214 110 142 10 4 4 517 72 KOH
215 45 100 1 10 na 5 72
224 20 110 1 5 5 01 10 5 72 KOH
229 110 11 5 10 55 73
230 3000
232 20 125 10 10
233 140 1 1 20 73 KOH
237 10 125 1 10 5 10 10 72 KOH
239 30 130 01 4 10 74
241 117 2 11 11 10 1 72
242 15 119 32 5 5 5 74 KOH
244 130 1 5 5 10 74 KOH
249 130 5 5 01 5 5 74 HCl
250 80 40 10 5 01 5 40 065 72 HCl
251 50 60 1 11 5 10 1 74 KOH
252 140 11 1 10 10 72 KOH
amphotericin-B
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
256 80 40 10 5 5 5 065 72
262 150 1 5 4 5 5 73 KOH
263^ 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
274 40 106 1 5 4 5 72 KOH
276 20 110 1 10 5 10 10 1 72 KOH
278 20 110 1 10 5 01 10 5 72 KOH
283 20 110 1 5 5 01 1 5 72 KOH
286 50 1 5 5 01 5 2 10 72 KOH
^phosphatidylinositol 45-diphosphate (5 mgml 1) was added to the internal solution to avoid the rundown of the current 70 L-aspartic acid 70 KOH
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Pipette solution of ItoID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phospho creatinne
NaCl CaCl2 KOH KH2PO4 pH pH buffer
47 49 50
130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
214 110 142 10 4 10 517 72 KOH
215 45 100 1 10 na 5 72
220 25 130 1 1 5 01 5 3 725 KOH
221 130 1 5 4 01 10 72 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245 20 110 5 5 10 72
256 80 40 10 5 5 5 065 60 72
263 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
Summary of IKur (Kv1531) protocolsbull Distribution a major repolarizing current in human atria but seems to play no
role in the ventricle bull Pharmacology selective IKur blockers to restore and maintain sinus rhythm
without a risk of ventricular pro-arrhythmia
bull Channel sources1 Transfected cell lines CHO L cells HEK 293 and Xenopus Oocyte (710 =
70)2 Myocyte (30) species ndash human AM and mouse VM
bull Temperature RT = 100bull Pulse sequences
ndash Holding range -80 to -50 mV (mode -80 mV)ndash Activation 20 - 60 mV (mode 50 mV)ndash Duration of activation 150 ndash 1000 ms (mode 300 ms)
1 IKur measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKurID Channel
CurrentSpeciescells CPD Temp Vh
(mV)Prepulse Activation
(ms mV)Tail current
Voltage -Dependence(ms mV)
Interval (s)
49 hKv15 MouseL cell vanoxerine RT -80 300 20 10s
221 Kv15 HEK293 RSD1235 25 -80 200 60
224 hKv15 HEK293 Allitridi 22-23 -80 300 50 300 -40 to 60
245-1 hKv15 CHO DPO RT -80 150 40 na -30
245-2 Ikur HumanAM DPO RT -50 300 40 na -30
245-3 hKv15 XO DPO na -80 na 40
262 hKv15 CHO SSR149744C RT -70 1000 30 20s
263 hKv15 Mouse LtK- Spironolactonecanrenoic acid
RT -80 500 60 500 -40 500 -80 to 60
263 Ikur MouseVM Spironolactonecanrenoic acid
RT -80 200 40 500 50 10s
283 Ikur HumanAM clotrimazole 21-22 -50 100 40 150 50 na -30 150 -40 to 50
Ikur (Kv15)
Extracellular buffer solution of IKurID NaCl NMCG K-
gluconate KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-acetate Blocker pH pH
buffer
49 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 10 033 CdCl2 (01) BaCl2 (02) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245-1 132 4 18 12 10 111 72 na
245-2 132 4 12 10 111 nisoldipine (04)
245-3 116 4 10 725
262 140 54 2 2 15 10 74 NaOH
263 130 4 1 1 10 10 74 NaOH
281 96 2 18 1 5 74 NaOH
283 140 5 1 1 5 10 033BaCl2 (02 Ik1) CdCl2 (Ica)
74 NaOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Pipette solution of IKurID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phosphocreatinne
NaCl CaCl2 K-gluconate KH2PO4 pH pH
buffer
49 130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
221 130 1 5 4 01 10 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245-1 15 32 5 5 5 119 735
245-2 20 110 5 5 10 72
245-3 4 10 003 116 725
262 140 2 11 10 2 73 KOH
263 42 80 5 5 5 3 10 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Summary of Iks protocolsbull Distribution all cell types but less in midmyocardial myocytes
bull Pharmacology Atrial and ventricular repolarizationHeart rate dependentbull subunit KCNQ1 (channel forming) Kv71 KvLQT1bull subunit KCNE1 (accessary unit activation of KCNQ1) minK (minimum potassium
subunit)
bull Channel sources1 transfected cell lines CHO HEK 293 Cos-7 cells and Xenopus Oocyte (2136 = 58)2 Myocyte (42) species ndash Canine VM Guinea pig VM rabbit VM and mouse VM (mode
GP)
bull Temperature RT = 1629 55 physiological temperature = 45
bull Pulse sequencesndash Holding potential range = -90 to -30 (mode -80)ndash Depolarization potential 20-60 mV ndash Duration of depolarization 50 ms ndash 10s (mode 2s)ndash Repolarization potential -80 to -10 mV (mode -40 mV)ndash Duration of repolarization 05 ndash 5s (mode 2s)
bull IKs measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation (s mV)
deactivation(s mV)
Interval (s)
47 hKvLQT1hminK CHO RT -80 2 40 01 Hz
49 hKv711hKCNE1 CHO RT -80 2 20 05 -40 15s
50 hKv711hKCNE1 HEk293 RT -80 2 20 05 -40 15s
184 IKs canineepiardial cells 37 -50 2 40 na -30 na
214 Kv711minK HEK293 na -80 5 -60 to 60 5s -50 na
215 IKs caninemyocyte na -40 5 50
224 hKCNQ1hKCNE1 HEK293 22-23 -80 3 -60 to 60 (20) na -50 na
229 hCNE1-hKCNQ1 Transgenic mousemyocyte RT -75 20 -40(Na block) 2 60 2 -40 006 Hz
230 KCNQ1KCNE1 XenopusOocyte 20-22 -80 5 30 na -70
232 KCNQ1KCNE1 HEK293 36 -80 2 -80 to 110 (10) 2 -60 6s
233 KCNQ1KCNE1 CHO na -80 80 -90 3 20 1 -50 005Hz
237 KCNQ1 CHO na -80 na -80 to 60 1 -40
239 KCNQ1KCNE1 HEK293 RT -70 1 -70 to 60 na -40
241 KCNQ1 andor minK MouseCOS-7 22-25 -80 3 -120 to 60 (20) 2 -30
242 hKvLQT1hminK HEK293 19-36 -80 3 -50 to 60 (10) 5 -50
242 hKvLQT1hminK HEK293 19-36 -50 1 or 2 50 na -50 10s
244 KCNQ1 andor KCNE1 CHO na -90 3 -70 to 70 (10) na -60
245 IKs GPMyocyte 35 -50 1 50
249 KCNQ1KCNE1 HEK293 33-34 -40 1 -40 to 80 (10) na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation(s mV)
Voltage dependent(s mV)
Deactivation(s mV)
Interval (s)
251 IKs GPMyocyte 22-2535-37 -40 2 60 2 -40 to 80 (20) na -40
252 IKs GPMyocyte 35 -80 200 -50 3 60 2 -40 60s
252 hminK XenopusOocyte 35 -80 10 20 10 -40 to 60 2 -80
256 IKs Rabbitmyocyte 34 -50 3 -40 to 50 na
260 hKCNQ1hKCNE1 HEK293 RT -80 25 -70 to 100 (10) 25 -40 15s
262 IKs GPMyocyte 35 -80 1 40 5 -10
263 Kv71 and minK CHO RT -80 2 40 na -40
264 IKs GPmyocyte na -80 200 -40 05 50 na -40
267 IKs GPmyocyte na -90 na 0 na 40 01- 015 0-80 -90
273 IKs Caninemyocyte 37 -40 3 50 na -40
274 IKs GPmyocyte 36 -30 05 50 or 2 70 05 (2) -30 to 70 na -30
275 KCNQ1KCNE1 CHO 21 -80 1 40 01Hz
276 hKCNQ1hminK CHO 22 -80 6 40 na -80 005Hz
278 IKs Canine myocyte 35-37 -50 3 -50 to 50 2s -40
283 KCNQ1KCNE1 HEK293 21-22 -80 3 40 3 -80 to 40 na -40
286 IKs GPmyocyte 34-36 -50 005 30 005 ndash3 -50 to 30 na -50
287 Iks GPmyocyte 36 -80 na -40 2 60 2 -40 to 80 na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
47 49 50
137 4 18 1 10 10 74 NaOH
184 145 4 18 18 10 11 NaH2PO4 1 E4031 (5uM) Cadmium (300 uM) 74 NaOH
214 140 4 2 1 10 74 NaOH
215 144 5 56 25 5 Dextrose 11 Nifedipine (5 uM) E4031 (1 uM) 74 NaOH
224 140 54 18 1 10 10 NaH2PO4 033 73 NaOH
229 132 48 2 12 10 5 E4031 nisoldipine 74 NaOH
230 96 2 18 1 5 74 NaOH
232 140 54 18 1 5 55 74 NaOH
233 17 120 4 2 1 10 10 74 NaOH
237 130 5 18 1 10 Na-Acetate 28 74 NaOH
239 150 54 18 4 10 74 NaOH
241 135 5 25 12 5 10 74 NaOH
242 132 4 18 12 10 111 74 NaOH
244 140 4 18 12 55 11 74 NaOH
249 140 54 18 05 5 55 NaH2PO4 033 74 NaOH
250 140 54 18 1 5 10 74 NaOH
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
251 1 1 2 10 N-methly-d-glucamine 132
lanthanum chloride 005 mMcadmium chloride 02 mM 74 HCl
252 140 47 13 1 10 10 74 NaOH
256 09 5 N-methly-d-glucamine 149
Nisoldipine 3 uME4031 10 uM 74 HCl
260 140 54 18 1 5 10 NaH2PO4 03 73 NaOH
262 137 4 18 2 10 115 Pyrubic acid 5 Thiamine 01 74 NaOH
263 130 4 1 1 10 10 TTX 003 mM CoCl2 2 mM 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10 KH2PO4 12 MgSO4 12 NaHCO3 44
Nifetidine 5 uM 4-aminopyridine 3 mM 72 NaOH
274 140 1 5 10 Cd2+ 02 mM 74 NaOH
275 130 54 18 1 10 5 74 NaOH
278 136 54 1 1 5 Dextrose 10 NaH2PO4 033 E4031 5 uM 74 NaOH
283 140 5 1 1 5 10 NaH2PO4 033 74 NaOH
286 09 5 5 N-methly-d-glucamine 149
Nisoldipine 04 uME4031 5 uM 74 NaOH
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
49 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
214 110 142 10 4 4 517 72 KOH
215 45 100 1 10 na 5 72
224 20 110 1 5 5 01 10 5 72 KOH
229 110 11 5 10 55 73
230 3000
232 20 125 10 10
233 140 1 1 20 73 KOH
237 10 125 1 10 5 10 10 72 KOH
239 30 130 01 4 10 74
241 117 2 11 11 10 1 72
242 15 119 32 5 5 5 74 KOH
244 130 1 5 5 10 74 KOH
249 130 5 5 01 5 5 74 HCl
250 80 40 10 5 01 5 40 065 72 HCl
251 50 60 1 11 5 10 1 74 KOH
252 140 11 1 10 10 72 KOH
amphotericin-B
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
256 80 40 10 5 5 5 065 72
262 150 1 5 4 5 5 73 KOH
263^ 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
274 40 106 1 5 4 5 72 KOH
276 20 110 1 10 5 10 10 1 72 KOH
278 20 110 1 10 5 01 10 5 72 KOH
283 20 110 1 5 5 01 1 5 72 KOH
286 50 1 5 5 01 5 2 10 72 KOH
^phosphatidylinositol 45-diphosphate (5 mgml 1) was added to the internal solution to avoid the rundown of the current 70 L-aspartic acid 70 KOH
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Summary of IKur (Kv1531) protocolsbull Distribution a major repolarizing current in human atria but seems to play no
role in the ventricle bull Pharmacology selective IKur blockers to restore and maintain sinus rhythm
without a risk of ventricular pro-arrhythmia
bull Channel sources1 Transfected cell lines CHO L cells HEK 293 and Xenopus Oocyte (710 =
70)2 Myocyte (30) species ndash human AM and mouse VM
bull Temperature RT = 100bull Pulse sequences
ndash Holding range -80 to -50 mV (mode -80 mV)ndash Activation 20 - 60 mV (mode 50 mV)ndash Duration of activation 150 ndash 1000 ms (mode 300 ms)
1 IKur measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKurID Channel
CurrentSpeciescells CPD Temp Vh
(mV)Prepulse Activation
(ms mV)Tail current
Voltage -Dependence(ms mV)
Interval (s)
49 hKv15 MouseL cell vanoxerine RT -80 300 20 10s
221 Kv15 HEK293 RSD1235 25 -80 200 60
224 hKv15 HEK293 Allitridi 22-23 -80 300 50 300 -40 to 60
245-1 hKv15 CHO DPO RT -80 150 40 na -30
245-2 Ikur HumanAM DPO RT -50 300 40 na -30
245-3 hKv15 XO DPO na -80 na 40
262 hKv15 CHO SSR149744C RT -70 1000 30 20s
263 hKv15 Mouse LtK- Spironolactonecanrenoic acid
RT -80 500 60 500 -40 500 -80 to 60
263 Ikur MouseVM Spironolactonecanrenoic acid
RT -80 200 40 500 50 10s
283 Ikur HumanAM clotrimazole 21-22 -50 100 40 150 50 na -30 150 -40 to 50
Ikur (Kv15)
Extracellular buffer solution of IKurID NaCl NMCG K-
gluconate KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-acetate Blocker pH pH
buffer
49 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 10 033 CdCl2 (01) BaCl2 (02) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245-1 132 4 18 12 10 111 72 na
245-2 132 4 12 10 111 nisoldipine (04)
245-3 116 4 10 725
262 140 54 2 2 15 10 74 NaOH
263 130 4 1 1 10 10 74 NaOH
281 96 2 18 1 5 74 NaOH
283 140 5 1 1 5 10 033BaCl2 (02 Ik1) CdCl2 (Ica)
74 NaOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Pipette solution of IKurID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phosphocreatinne
NaCl CaCl2 K-gluconate KH2PO4 pH pH
buffer
49 130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
221 130 1 5 4 01 10 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245-1 15 32 5 5 5 119 735
245-2 20 110 5 5 10 72
245-3 4 10 003 116 725
262 140 2 11 10 2 73 KOH
263 42 80 5 5 5 3 10 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Summary of Iks protocolsbull Distribution all cell types but less in midmyocardial myocytes
bull Pharmacology Atrial and ventricular repolarizationHeart rate dependentbull subunit KCNQ1 (channel forming) Kv71 KvLQT1bull subunit KCNE1 (accessary unit activation of KCNQ1) minK (minimum potassium
subunit)
bull Channel sources1 transfected cell lines CHO HEK 293 Cos-7 cells and Xenopus Oocyte (2136 = 58)2 Myocyte (42) species ndash Canine VM Guinea pig VM rabbit VM and mouse VM (mode
GP)
bull Temperature RT = 1629 55 physiological temperature = 45
bull Pulse sequencesndash Holding potential range = -90 to -30 (mode -80)ndash Depolarization potential 20-60 mV ndash Duration of depolarization 50 ms ndash 10s (mode 2s)ndash Repolarization potential -80 to -10 mV (mode -40 mV)ndash Duration of repolarization 05 ndash 5s (mode 2s)
bull IKs measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation (s mV)
deactivation(s mV)
Interval (s)
47 hKvLQT1hminK CHO RT -80 2 40 01 Hz
49 hKv711hKCNE1 CHO RT -80 2 20 05 -40 15s
50 hKv711hKCNE1 HEk293 RT -80 2 20 05 -40 15s
184 IKs canineepiardial cells 37 -50 2 40 na -30 na
214 Kv711minK HEK293 na -80 5 -60 to 60 5s -50 na
215 IKs caninemyocyte na -40 5 50
224 hKCNQ1hKCNE1 HEK293 22-23 -80 3 -60 to 60 (20) na -50 na
229 hCNE1-hKCNQ1 Transgenic mousemyocyte RT -75 20 -40(Na block) 2 60 2 -40 006 Hz
230 KCNQ1KCNE1 XenopusOocyte 20-22 -80 5 30 na -70
232 KCNQ1KCNE1 HEK293 36 -80 2 -80 to 110 (10) 2 -60 6s
233 KCNQ1KCNE1 CHO na -80 80 -90 3 20 1 -50 005Hz
237 KCNQ1 CHO na -80 na -80 to 60 1 -40
239 KCNQ1KCNE1 HEK293 RT -70 1 -70 to 60 na -40
241 KCNQ1 andor minK MouseCOS-7 22-25 -80 3 -120 to 60 (20) 2 -30
242 hKvLQT1hminK HEK293 19-36 -80 3 -50 to 60 (10) 5 -50
242 hKvLQT1hminK HEK293 19-36 -50 1 or 2 50 na -50 10s
244 KCNQ1 andor KCNE1 CHO na -90 3 -70 to 70 (10) na -60
245 IKs GPMyocyte 35 -50 1 50
249 KCNQ1KCNE1 HEK293 33-34 -40 1 -40 to 80 (10) na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation(s mV)
Voltage dependent(s mV)
Deactivation(s mV)
Interval (s)
251 IKs GPMyocyte 22-2535-37 -40 2 60 2 -40 to 80 (20) na -40
252 IKs GPMyocyte 35 -80 200 -50 3 60 2 -40 60s
252 hminK XenopusOocyte 35 -80 10 20 10 -40 to 60 2 -80
256 IKs Rabbitmyocyte 34 -50 3 -40 to 50 na
260 hKCNQ1hKCNE1 HEK293 RT -80 25 -70 to 100 (10) 25 -40 15s
262 IKs GPMyocyte 35 -80 1 40 5 -10
263 Kv71 and minK CHO RT -80 2 40 na -40
264 IKs GPmyocyte na -80 200 -40 05 50 na -40
267 IKs GPmyocyte na -90 na 0 na 40 01- 015 0-80 -90
273 IKs Caninemyocyte 37 -40 3 50 na -40
274 IKs GPmyocyte 36 -30 05 50 or 2 70 05 (2) -30 to 70 na -30
275 KCNQ1KCNE1 CHO 21 -80 1 40 01Hz
276 hKCNQ1hminK CHO 22 -80 6 40 na -80 005Hz
278 IKs Canine myocyte 35-37 -50 3 -50 to 50 2s -40
283 KCNQ1KCNE1 HEK293 21-22 -80 3 40 3 -80 to 40 na -40
286 IKs GPmyocyte 34-36 -50 005 30 005 ndash3 -50 to 30 na -50
287 Iks GPmyocyte 36 -80 na -40 2 60 2 -40 to 80 na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
47 49 50
137 4 18 1 10 10 74 NaOH
184 145 4 18 18 10 11 NaH2PO4 1 E4031 (5uM) Cadmium (300 uM) 74 NaOH
214 140 4 2 1 10 74 NaOH
215 144 5 56 25 5 Dextrose 11 Nifedipine (5 uM) E4031 (1 uM) 74 NaOH
224 140 54 18 1 10 10 NaH2PO4 033 73 NaOH
229 132 48 2 12 10 5 E4031 nisoldipine 74 NaOH
230 96 2 18 1 5 74 NaOH
232 140 54 18 1 5 55 74 NaOH
233 17 120 4 2 1 10 10 74 NaOH
237 130 5 18 1 10 Na-Acetate 28 74 NaOH
239 150 54 18 4 10 74 NaOH
241 135 5 25 12 5 10 74 NaOH
242 132 4 18 12 10 111 74 NaOH
244 140 4 18 12 55 11 74 NaOH
249 140 54 18 05 5 55 NaH2PO4 033 74 NaOH
250 140 54 18 1 5 10 74 NaOH
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
251 1 1 2 10 N-methly-d-glucamine 132
lanthanum chloride 005 mMcadmium chloride 02 mM 74 HCl
252 140 47 13 1 10 10 74 NaOH
256 09 5 N-methly-d-glucamine 149
Nisoldipine 3 uME4031 10 uM 74 HCl
260 140 54 18 1 5 10 NaH2PO4 03 73 NaOH
262 137 4 18 2 10 115 Pyrubic acid 5 Thiamine 01 74 NaOH
263 130 4 1 1 10 10 TTX 003 mM CoCl2 2 mM 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10 KH2PO4 12 MgSO4 12 NaHCO3 44
Nifetidine 5 uM 4-aminopyridine 3 mM 72 NaOH
274 140 1 5 10 Cd2+ 02 mM 74 NaOH
275 130 54 18 1 10 5 74 NaOH
278 136 54 1 1 5 Dextrose 10 NaH2PO4 033 E4031 5 uM 74 NaOH
283 140 5 1 1 5 10 NaH2PO4 033 74 NaOH
286 09 5 5 N-methly-d-glucamine 149
Nisoldipine 04 uME4031 5 uM 74 NaOH
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
49 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
214 110 142 10 4 4 517 72 KOH
215 45 100 1 10 na 5 72
224 20 110 1 5 5 01 10 5 72 KOH
229 110 11 5 10 55 73
230 3000
232 20 125 10 10
233 140 1 1 20 73 KOH
237 10 125 1 10 5 10 10 72 KOH
239 30 130 01 4 10 74
241 117 2 11 11 10 1 72
242 15 119 32 5 5 5 74 KOH
244 130 1 5 5 10 74 KOH
249 130 5 5 01 5 5 74 HCl
250 80 40 10 5 01 5 40 065 72 HCl
251 50 60 1 11 5 10 1 74 KOH
252 140 11 1 10 10 72 KOH
amphotericin-B
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
256 80 40 10 5 5 5 065 72
262 150 1 5 4 5 5 73 KOH
263^ 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
274 40 106 1 5 4 5 72 KOH
276 20 110 1 10 5 10 10 1 72 KOH
278 20 110 1 10 5 01 10 5 72 KOH
283 20 110 1 5 5 01 1 5 72 KOH
286 50 1 5 5 01 5 2 10 72 KOH
^phosphatidylinositol 45-diphosphate (5 mgml 1) was added to the internal solution to avoid the rundown of the current 70 L-aspartic acid 70 KOH
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Pulse sequences IKurID Channel
CurrentSpeciescells CPD Temp Vh
(mV)Prepulse Activation
(ms mV)Tail current
Voltage -Dependence(ms mV)
Interval (s)
49 hKv15 MouseL cell vanoxerine RT -80 300 20 10s
221 Kv15 HEK293 RSD1235 25 -80 200 60
224 hKv15 HEK293 Allitridi 22-23 -80 300 50 300 -40 to 60
245-1 hKv15 CHO DPO RT -80 150 40 na -30
245-2 Ikur HumanAM DPO RT -50 300 40 na -30
245-3 hKv15 XO DPO na -80 na 40
262 hKv15 CHO SSR149744C RT -70 1000 30 20s
263 hKv15 Mouse LtK- Spironolactonecanrenoic acid
RT -80 500 60 500 -40 500 -80 to 60
263 Ikur MouseVM Spironolactonecanrenoic acid
RT -80 200 40 500 50 10s
283 Ikur HumanAM clotrimazole 21-22 -50 100 40 150 50 na -30 150 -40 to 50
Ikur (Kv15)
Extracellular buffer solution of IKurID NaCl NMCG K-
gluconate KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-acetate Blocker pH pH
buffer
49 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 10 033 CdCl2 (01) BaCl2 (02) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245-1 132 4 18 12 10 111 72 na
245-2 132 4 12 10 111 nisoldipine (04)
245-3 116 4 10 725
262 140 54 2 2 15 10 74 NaOH
263 130 4 1 1 10 10 74 NaOH
281 96 2 18 1 5 74 NaOH
283 140 5 1 1 5 10 033BaCl2 (02 Ik1) CdCl2 (Ica)
74 NaOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Pipette solution of IKurID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phosphocreatinne
NaCl CaCl2 K-gluconate KH2PO4 pH pH
buffer
49 130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
221 130 1 5 4 01 10 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245-1 15 32 5 5 5 119 735
245-2 20 110 5 5 10 72
245-3 4 10 003 116 725
262 140 2 11 10 2 73 KOH
263 42 80 5 5 5 3 10 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Summary of Iks protocolsbull Distribution all cell types but less in midmyocardial myocytes
bull Pharmacology Atrial and ventricular repolarizationHeart rate dependentbull subunit KCNQ1 (channel forming) Kv71 KvLQT1bull subunit KCNE1 (accessary unit activation of KCNQ1) minK (minimum potassium
subunit)
bull Channel sources1 transfected cell lines CHO HEK 293 Cos-7 cells and Xenopus Oocyte (2136 = 58)2 Myocyte (42) species ndash Canine VM Guinea pig VM rabbit VM and mouse VM (mode
GP)
bull Temperature RT = 1629 55 physiological temperature = 45
bull Pulse sequencesndash Holding potential range = -90 to -30 (mode -80)ndash Depolarization potential 20-60 mV ndash Duration of depolarization 50 ms ndash 10s (mode 2s)ndash Repolarization potential -80 to -10 mV (mode -40 mV)ndash Duration of repolarization 05 ndash 5s (mode 2s)
bull IKs measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation (s mV)
deactivation(s mV)
Interval (s)
47 hKvLQT1hminK CHO RT -80 2 40 01 Hz
49 hKv711hKCNE1 CHO RT -80 2 20 05 -40 15s
50 hKv711hKCNE1 HEk293 RT -80 2 20 05 -40 15s
184 IKs canineepiardial cells 37 -50 2 40 na -30 na
214 Kv711minK HEK293 na -80 5 -60 to 60 5s -50 na
215 IKs caninemyocyte na -40 5 50
224 hKCNQ1hKCNE1 HEK293 22-23 -80 3 -60 to 60 (20) na -50 na
229 hCNE1-hKCNQ1 Transgenic mousemyocyte RT -75 20 -40(Na block) 2 60 2 -40 006 Hz
230 KCNQ1KCNE1 XenopusOocyte 20-22 -80 5 30 na -70
232 KCNQ1KCNE1 HEK293 36 -80 2 -80 to 110 (10) 2 -60 6s
233 KCNQ1KCNE1 CHO na -80 80 -90 3 20 1 -50 005Hz
237 KCNQ1 CHO na -80 na -80 to 60 1 -40
239 KCNQ1KCNE1 HEK293 RT -70 1 -70 to 60 na -40
241 KCNQ1 andor minK MouseCOS-7 22-25 -80 3 -120 to 60 (20) 2 -30
242 hKvLQT1hminK HEK293 19-36 -80 3 -50 to 60 (10) 5 -50
242 hKvLQT1hminK HEK293 19-36 -50 1 or 2 50 na -50 10s
244 KCNQ1 andor KCNE1 CHO na -90 3 -70 to 70 (10) na -60
245 IKs GPMyocyte 35 -50 1 50
249 KCNQ1KCNE1 HEK293 33-34 -40 1 -40 to 80 (10) na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation(s mV)
Voltage dependent(s mV)
Deactivation(s mV)
Interval (s)
251 IKs GPMyocyte 22-2535-37 -40 2 60 2 -40 to 80 (20) na -40
252 IKs GPMyocyte 35 -80 200 -50 3 60 2 -40 60s
252 hminK XenopusOocyte 35 -80 10 20 10 -40 to 60 2 -80
256 IKs Rabbitmyocyte 34 -50 3 -40 to 50 na
260 hKCNQ1hKCNE1 HEK293 RT -80 25 -70 to 100 (10) 25 -40 15s
262 IKs GPMyocyte 35 -80 1 40 5 -10
263 Kv71 and minK CHO RT -80 2 40 na -40
264 IKs GPmyocyte na -80 200 -40 05 50 na -40
267 IKs GPmyocyte na -90 na 0 na 40 01- 015 0-80 -90
273 IKs Caninemyocyte 37 -40 3 50 na -40
274 IKs GPmyocyte 36 -30 05 50 or 2 70 05 (2) -30 to 70 na -30
275 KCNQ1KCNE1 CHO 21 -80 1 40 01Hz
276 hKCNQ1hminK CHO 22 -80 6 40 na -80 005Hz
278 IKs Canine myocyte 35-37 -50 3 -50 to 50 2s -40
283 KCNQ1KCNE1 HEK293 21-22 -80 3 40 3 -80 to 40 na -40
286 IKs GPmyocyte 34-36 -50 005 30 005 ndash3 -50 to 30 na -50
287 Iks GPmyocyte 36 -80 na -40 2 60 2 -40 to 80 na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
47 49 50
137 4 18 1 10 10 74 NaOH
184 145 4 18 18 10 11 NaH2PO4 1 E4031 (5uM) Cadmium (300 uM) 74 NaOH
214 140 4 2 1 10 74 NaOH
215 144 5 56 25 5 Dextrose 11 Nifedipine (5 uM) E4031 (1 uM) 74 NaOH
224 140 54 18 1 10 10 NaH2PO4 033 73 NaOH
229 132 48 2 12 10 5 E4031 nisoldipine 74 NaOH
230 96 2 18 1 5 74 NaOH
232 140 54 18 1 5 55 74 NaOH
233 17 120 4 2 1 10 10 74 NaOH
237 130 5 18 1 10 Na-Acetate 28 74 NaOH
239 150 54 18 4 10 74 NaOH
241 135 5 25 12 5 10 74 NaOH
242 132 4 18 12 10 111 74 NaOH
244 140 4 18 12 55 11 74 NaOH
249 140 54 18 05 5 55 NaH2PO4 033 74 NaOH
250 140 54 18 1 5 10 74 NaOH
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
251 1 1 2 10 N-methly-d-glucamine 132
lanthanum chloride 005 mMcadmium chloride 02 mM 74 HCl
252 140 47 13 1 10 10 74 NaOH
256 09 5 N-methly-d-glucamine 149
Nisoldipine 3 uME4031 10 uM 74 HCl
260 140 54 18 1 5 10 NaH2PO4 03 73 NaOH
262 137 4 18 2 10 115 Pyrubic acid 5 Thiamine 01 74 NaOH
263 130 4 1 1 10 10 TTX 003 mM CoCl2 2 mM 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10 KH2PO4 12 MgSO4 12 NaHCO3 44
Nifetidine 5 uM 4-aminopyridine 3 mM 72 NaOH
274 140 1 5 10 Cd2+ 02 mM 74 NaOH
275 130 54 18 1 10 5 74 NaOH
278 136 54 1 1 5 Dextrose 10 NaH2PO4 033 E4031 5 uM 74 NaOH
283 140 5 1 1 5 10 NaH2PO4 033 74 NaOH
286 09 5 5 N-methly-d-glucamine 149
Nisoldipine 04 uME4031 5 uM 74 NaOH
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
49 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
214 110 142 10 4 4 517 72 KOH
215 45 100 1 10 na 5 72
224 20 110 1 5 5 01 10 5 72 KOH
229 110 11 5 10 55 73
230 3000
232 20 125 10 10
233 140 1 1 20 73 KOH
237 10 125 1 10 5 10 10 72 KOH
239 30 130 01 4 10 74
241 117 2 11 11 10 1 72
242 15 119 32 5 5 5 74 KOH
244 130 1 5 5 10 74 KOH
249 130 5 5 01 5 5 74 HCl
250 80 40 10 5 01 5 40 065 72 HCl
251 50 60 1 11 5 10 1 74 KOH
252 140 11 1 10 10 72 KOH
amphotericin-B
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
256 80 40 10 5 5 5 065 72
262 150 1 5 4 5 5 73 KOH
263^ 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
274 40 106 1 5 4 5 72 KOH
276 20 110 1 10 5 10 10 1 72 KOH
278 20 110 1 10 5 01 10 5 72 KOH
283 20 110 1 5 5 01 1 5 72 KOH
286 50 1 5 5 01 5 2 10 72 KOH
^phosphatidylinositol 45-diphosphate (5 mgml 1) was added to the internal solution to avoid the rundown of the current 70 L-aspartic acid 70 KOH
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Extracellular buffer solution of IKurID NaCl NMCG K-
gluconate KCl CaCl2 MgCl2 HEPES glucose dextrose NaH2PO4 Na-acetate Blocker pH pH
buffer
49 137 4 18 1 10 10 74 NaOH
206 135 54 18 1 10 10 033 CdCl2 (01) BaCl2 (02) 74 NaOH
210 137 4 18 12 10 11 CdCl2 (02) 74 NaOH
221 135 5 1 1 10 28 74 NaOH
224 140 54 18 1 10 10 033 73 NaOH
245-1 132 4 18 12 10 111 72 na
245-2 132 4 12 10 111 nisoldipine (04)
245-3 116 4 10 725
262 140 54 2 2 15 10 74 NaOH
263 130 4 1 1 10 10 74 NaOH
281 96 2 18 1 5 74 NaOH
283 140 5 1 1 5 10 033BaCl2 (02 Ik1) CdCl2 (Ica)
74 NaOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Pipette solution of IKurID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phosphocreatinne
NaCl CaCl2 K-gluconate KH2PO4 pH pH
buffer
49 130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
221 130 1 5 4 01 10 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245-1 15 32 5 5 5 119 735
245-2 20 110 5 5 10 72
245-3 4 10 003 116 725
262 140 2 11 10 2 73 KOH
263 42 80 5 5 5 3 10 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Summary of Iks protocolsbull Distribution all cell types but less in midmyocardial myocytes
bull Pharmacology Atrial and ventricular repolarizationHeart rate dependentbull subunit KCNQ1 (channel forming) Kv71 KvLQT1bull subunit KCNE1 (accessary unit activation of KCNQ1) minK (minimum potassium
subunit)
bull Channel sources1 transfected cell lines CHO HEK 293 Cos-7 cells and Xenopus Oocyte (2136 = 58)2 Myocyte (42) species ndash Canine VM Guinea pig VM rabbit VM and mouse VM (mode
GP)
bull Temperature RT = 1629 55 physiological temperature = 45
bull Pulse sequencesndash Holding potential range = -90 to -30 (mode -80)ndash Depolarization potential 20-60 mV ndash Duration of depolarization 50 ms ndash 10s (mode 2s)ndash Repolarization potential -80 to -10 mV (mode -40 mV)ndash Duration of repolarization 05 ndash 5s (mode 2s)
bull IKs measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation (s mV)
deactivation(s mV)
Interval (s)
47 hKvLQT1hminK CHO RT -80 2 40 01 Hz
49 hKv711hKCNE1 CHO RT -80 2 20 05 -40 15s
50 hKv711hKCNE1 HEk293 RT -80 2 20 05 -40 15s
184 IKs canineepiardial cells 37 -50 2 40 na -30 na
214 Kv711minK HEK293 na -80 5 -60 to 60 5s -50 na
215 IKs caninemyocyte na -40 5 50
224 hKCNQ1hKCNE1 HEK293 22-23 -80 3 -60 to 60 (20) na -50 na
229 hCNE1-hKCNQ1 Transgenic mousemyocyte RT -75 20 -40(Na block) 2 60 2 -40 006 Hz
230 KCNQ1KCNE1 XenopusOocyte 20-22 -80 5 30 na -70
232 KCNQ1KCNE1 HEK293 36 -80 2 -80 to 110 (10) 2 -60 6s
233 KCNQ1KCNE1 CHO na -80 80 -90 3 20 1 -50 005Hz
237 KCNQ1 CHO na -80 na -80 to 60 1 -40
239 KCNQ1KCNE1 HEK293 RT -70 1 -70 to 60 na -40
241 KCNQ1 andor minK MouseCOS-7 22-25 -80 3 -120 to 60 (20) 2 -30
242 hKvLQT1hminK HEK293 19-36 -80 3 -50 to 60 (10) 5 -50
242 hKvLQT1hminK HEK293 19-36 -50 1 or 2 50 na -50 10s
244 KCNQ1 andor KCNE1 CHO na -90 3 -70 to 70 (10) na -60
245 IKs GPMyocyte 35 -50 1 50
249 KCNQ1KCNE1 HEK293 33-34 -40 1 -40 to 80 (10) na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation(s mV)
Voltage dependent(s mV)
Deactivation(s mV)
Interval (s)
251 IKs GPMyocyte 22-2535-37 -40 2 60 2 -40 to 80 (20) na -40
252 IKs GPMyocyte 35 -80 200 -50 3 60 2 -40 60s
252 hminK XenopusOocyte 35 -80 10 20 10 -40 to 60 2 -80
256 IKs Rabbitmyocyte 34 -50 3 -40 to 50 na
260 hKCNQ1hKCNE1 HEK293 RT -80 25 -70 to 100 (10) 25 -40 15s
262 IKs GPMyocyte 35 -80 1 40 5 -10
263 Kv71 and minK CHO RT -80 2 40 na -40
264 IKs GPmyocyte na -80 200 -40 05 50 na -40
267 IKs GPmyocyte na -90 na 0 na 40 01- 015 0-80 -90
273 IKs Caninemyocyte 37 -40 3 50 na -40
274 IKs GPmyocyte 36 -30 05 50 or 2 70 05 (2) -30 to 70 na -30
275 KCNQ1KCNE1 CHO 21 -80 1 40 01Hz
276 hKCNQ1hminK CHO 22 -80 6 40 na -80 005Hz
278 IKs Canine myocyte 35-37 -50 3 -50 to 50 2s -40
283 KCNQ1KCNE1 HEK293 21-22 -80 3 40 3 -80 to 40 na -40
286 IKs GPmyocyte 34-36 -50 005 30 005 ndash3 -50 to 30 na -50
287 Iks GPmyocyte 36 -80 na -40 2 60 2 -40 to 80 na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
47 49 50
137 4 18 1 10 10 74 NaOH
184 145 4 18 18 10 11 NaH2PO4 1 E4031 (5uM) Cadmium (300 uM) 74 NaOH
214 140 4 2 1 10 74 NaOH
215 144 5 56 25 5 Dextrose 11 Nifedipine (5 uM) E4031 (1 uM) 74 NaOH
224 140 54 18 1 10 10 NaH2PO4 033 73 NaOH
229 132 48 2 12 10 5 E4031 nisoldipine 74 NaOH
230 96 2 18 1 5 74 NaOH
232 140 54 18 1 5 55 74 NaOH
233 17 120 4 2 1 10 10 74 NaOH
237 130 5 18 1 10 Na-Acetate 28 74 NaOH
239 150 54 18 4 10 74 NaOH
241 135 5 25 12 5 10 74 NaOH
242 132 4 18 12 10 111 74 NaOH
244 140 4 18 12 55 11 74 NaOH
249 140 54 18 05 5 55 NaH2PO4 033 74 NaOH
250 140 54 18 1 5 10 74 NaOH
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
251 1 1 2 10 N-methly-d-glucamine 132
lanthanum chloride 005 mMcadmium chloride 02 mM 74 HCl
252 140 47 13 1 10 10 74 NaOH
256 09 5 N-methly-d-glucamine 149
Nisoldipine 3 uME4031 10 uM 74 HCl
260 140 54 18 1 5 10 NaH2PO4 03 73 NaOH
262 137 4 18 2 10 115 Pyrubic acid 5 Thiamine 01 74 NaOH
263 130 4 1 1 10 10 TTX 003 mM CoCl2 2 mM 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10 KH2PO4 12 MgSO4 12 NaHCO3 44
Nifetidine 5 uM 4-aminopyridine 3 mM 72 NaOH
274 140 1 5 10 Cd2+ 02 mM 74 NaOH
275 130 54 18 1 10 5 74 NaOH
278 136 54 1 1 5 Dextrose 10 NaH2PO4 033 E4031 5 uM 74 NaOH
283 140 5 1 1 5 10 NaH2PO4 033 74 NaOH
286 09 5 5 N-methly-d-glucamine 149
Nisoldipine 04 uME4031 5 uM 74 NaOH
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
49 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
214 110 142 10 4 4 517 72 KOH
215 45 100 1 10 na 5 72
224 20 110 1 5 5 01 10 5 72 KOH
229 110 11 5 10 55 73
230 3000
232 20 125 10 10
233 140 1 1 20 73 KOH
237 10 125 1 10 5 10 10 72 KOH
239 30 130 01 4 10 74
241 117 2 11 11 10 1 72
242 15 119 32 5 5 5 74 KOH
244 130 1 5 5 10 74 KOH
249 130 5 5 01 5 5 74 HCl
250 80 40 10 5 01 5 40 065 72 HCl
251 50 60 1 11 5 10 1 74 KOH
252 140 11 1 10 10 72 KOH
amphotericin-B
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
256 80 40 10 5 5 5 065 72
262 150 1 5 4 5 5 73 KOH
263^ 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
274 40 106 1 5 4 5 72 KOH
276 20 110 1 10 5 10 10 1 72 KOH
278 20 110 1 10 5 01 10 5 72 KOH
283 20 110 1 5 5 01 1 5 72 KOH
286 50 1 5 5 01 5 2 10 72 KOH
^phosphatidylinositol 45-diphosphate (5 mgml 1) was added to the internal solution to avoid the rundown of the current 70 L-aspartic acid 70 KOH
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Pipette solution of IKurID KCl K-
aspartate MgCl2 EGTA ATP GTP HEPESNa2-
phosphocreatinne
NaCl CaCl2 K-gluconate KH2PO4 pH pH
buffer
49 130 5 5 4 10 72 KOH
206 150 1 5 3 5 73 KOH
210 130 1 5 5 5 7 72 KOH
221 130 1 5 4 01 10 72 KOH
224 20 110 1 5 5 01 10 5 72 KOH
245-1 15 32 5 5 5 119 735
245-2 20 110 5 5 10 72
245-3 4 10 003 116 725
262 140 2 11 10 2 73 KOH
263 42 80 5 5 5 3 10 72 KOH
283 20 110 1 5 5 01 10 5 72 KOH
245-1 CHO 245-2 human AM 245-3 XO (inside out VC)
Summary of Iks protocolsbull Distribution all cell types but less in midmyocardial myocytes
bull Pharmacology Atrial and ventricular repolarizationHeart rate dependentbull subunit KCNQ1 (channel forming) Kv71 KvLQT1bull subunit KCNE1 (accessary unit activation of KCNQ1) minK (minimum potassium
subunit)
bull Channel sources1 transfected cell lines CHO HEK 293 Cos-7 cells and Xenopus Oocyte (2136 = 58)2 Myocyte (42) species ndash Canine VM Guinea pig VM rabbit VM and mouse VM (mode
GP)
bull Temperature RT = 1629 55 physiological temperature = 45
bull Pulse sequencesndash Holding potential range = -90 to -30 (mode -80)ndash Depolarization potential 20-60 mV ndash Duration of depolarization 50 ms ndash 10s (mode 2s)ndash Repolarization potential -80 to -10 mV (mode -40 mV)ndash Duration of repolarization 05 ndash 5s (mode 2s)
bull IKs measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation (s mV)
deactivation(s mV)
Interval (s)
47 hKvLQT1hminK CHO RT -80 2 40 01 Hz
49 hKv711hKCNE1 CHO RT -80 2 20 05 -40 15s
50 hKv711hKCNE1 HEk293 RT -80 2 20 05 -40 15s
184 IKs canineepiardial cells 37 -50 2 40 na -30 na
214 Kv711minK HEK293 na -80 5 -60 to 60 5s -50 na
215 IKs caninemyocyte na -40 5 50
224 hKCNQ1hKCNE1 HEK293 22-23 -80 3 -60 to 60 (20) na -50 na
229 hCNE1-hKCNQ1 Transgenic mousemyocyte RT -75 20 -40(Na block) 2 60 2 -40 006 Hz
230 KCNQ1KCNE1 XenopusOocyte 20-22 -80 5 30 na -70
232 KCNQ1KCNE1 HEK293 36 -80 2 -80 to 110 (10) 2 -60 6s
233 KCNQ1KCNE1 CHO na -80 80 -90 3 20 1 -50 005Hz
237 KCNQ1 CHO na -80 na -80 to 60 1 -40
239 KCNQ1KCNE1 HEK293 RT -70 1 -70 to 60 na -40
241 KCNQ1 andor minK MouseCOS-7 22-25 -80 3 -120 to 60 (20) 2 -30
242 hKvLQT1hminK HEK293 19-36 -80 3 -50 to 60 (10) 5 -50
242 hKvLQT1hminK HEK293 19-36 -50 1 or 2 50 na -50 10s
244 KCNQ1 andor KCNE1 CHO na -90 3 -70 to 70 (10) na -60
245 IKs GPMyocyte 35 -50 1 50
249 KCNQ1KCNE1 HEK293 33-34 -40 1 -40 to 80 (10) na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation(s mV)
Voltage dependent(s mV)
Deactivation(s mV)
Interval (s)
251 IKs GPMyocyte 22-2535-37 -40 2 60 2 -40 to 80 (20) na -40
252 IKs GPMyocyte 35 -80 200 -50 3 60 2 -40 60s
252 hminK XenopusOocyte 35 -80 10 20 10 -40 to 60 2 -80
256 IKs Rabbitmyocyte 34 -50 3 -40 to 50 na
260 hKCNQ1hKCNE1 HEK293 RT -80 25 -70 to 100 (10) 25 -40 15s
262 IKs GPMyocyte 35 -80 1 40 5 -10
263 Kv71 and minK CHO RT -80 2 40 na -40
264 IKs GPmyocyte na -80 200 -40 05 50 na -40
267 IKs GPmyocyte na -90 na 0 na 40 01- 015 0-80 -90
273 IKs Caninemyocyte 37 -40 3 50 na -40
274 IKs GPmyocyte 36 -30 05 50 or 2 70 05 (2) -30 to 70 na -30
275 KCNQ1KCNE1 CHO 21 -80 1 40 01Hz
276 hKCNQ1hminK CHO 22 -80 6 40 na -80 005Hz
278 IKs Canine myocyte 35-37 -50 3 -50 to 50 2s -40
283 KCNQ1KCNE1 HEK293 21-22 -80 3 40 3 -80 to 40 na -40
286 IKs GPmyocyte 34-36 -50 005 30 005 ndash3 -50 to 30 na -50
287 Iks GPmyocyte 36 -80 na -40 2 60 2 -40 to 80 na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
47 49 50
137 4 18 1 10 10 74 NaOH
184 145 4 18 18 10 11 NaH2PO4 1 E4031 (5uM) Cadmium (300 uM) 74 NaOH
214 140 4 2 1 10 74 NaOH
215 144 5 56 25 5 Dextrose 11 Nifedipine (5 uM) E4031 (1 uM) 74 NaOH
224 140 54 18 1 10 10 NaH2PO4 033 73 NaOH
229 132 48 2 12 10 5 E4031 nisoldipine 74 NaOH
230 96 2 18 1 5 74 NaOH
232 140 54 18 1 5 55 74 NaOH
233 17 120 4 2 1 10 10 74 NaOH
237 130 5 18 1 10 Na-Acetate 28 74 NaOH
239 150 54 18 4 10 74 NaOH
241 135 5 25 12 5 10 74 NaOH
242 132 4 18 12 10 111 74 NaOH
244 140 4 18 12 55 11 74 NaOH
249 140 54 18 05 5 55 NaH2PO4 033 74 NaOH
250 140 54 18 1 5 10 74 NaOH
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
251 1 1 2 10 N-methly-d-glucamine 132
lanthanum chloride 005 mMcadmium chloride 02 mM 74 HCl
252 140 47 13 1 10 10 74 NaOH
256 09 5 N-methly-d-glucamine 149
Nisoldipine 3 uME4031 10 uM 74 HCl
260 140 54 18 1 5 10 NaH2PO4 03 73 NaOH
262 137 4 18 2 10 115 Pyrubic acid 5 Thiamine 01 74 NaOH
263 130 4 1 1 10 10 TTX 003 mM CoCl2 2 mM 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10 KH2PO4 12 MgSO4 12 NaHCO3 44
Nifetidine 5 uM 4-aminopyridine 3 mM 72 NaOH
274 140 1 5 10 Cd2+ 02 mM 74 NaOH
275 130 54 18 1 10 5 74 NaOH
278 136 54 1 1 5 Dextrose 10 NaH2PO4 033 E4031 5 uM 74 NaOH
283 140 5 1 1 5 10 NaH2PO4 033 74 NaOH
286 09 5 5 N-methly-d-glucamine 149
Nisoldipine 04 uME4031 5 uM 74 NaOH
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
49 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
214 110 142 10 4 4 517 72 KOH
215 45 100 1 10 na 5 72
224 20 110 1 5 5 01 10 5 72 KOH
229 110 11 5 10 55 73
230 3000
232 20 125 10 10
233 140 1 1 20 73 KOH
237 10 125 1 10 5 10 10 72 KOH
239 30 130 01 4 10 74
241 117 2 11 11 10 1 72
242 15 119 32 5 5 5 74 KOH
244 130 1 5 5 10 74 KOH
249 130 5 5 01 5 5 74 HCl
250 80 40 10 5 01 5 40 065 72 HCl
251 50 60 1 11 5 10 1 74 KOH
252 140 11 1 10 10 72 KOH
amphotericin-B
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
256 80 40 10 5 5 5 065 72
262 150 1 5 4 5 5 73 KOH
263^ 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
274 40 106 1 5 4 5 72 KOH
276 20 110 1 10 5 10 10 1 72 KOH
278 20 110 1 10 5 01 10 5 72 KOH
283 20 110 1 5 5 01 1 5 72 KOH
286 50 1 5 5 01 5 2 10 72 KOH
^phosphatidylinositol 45-diphosphate (5 mgml 1) was added to the internal solution to avoid the rundown of the current 70 L-aspartic acid 70 KOH
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Summary of Iks protocolsbull Distribution all cell types but less in midmyocardial myocytes
bull Pharmacology Atrial and ventricular repolarizationHeart rate dependentbull subunit KCNQ1 (channel forming) Kv71 KvLQT1bull subunit KCNE1 (accessary unit activation of KCNQ1) minK (minimum potassium
subunit)
bull Channel sources1 transfected cell lines CHO HEK 293 Cos-7 cells and Xenopus Oocyte (2136 = 58)2 Myocyte (42) species ndash Canine VM Guinea pig VM rabbit VM and mouse VM (mode
GP)
bull Temperature RT = 1629 55 physiological temperature = 45
bull Pulse sequencesndash Holding potential range = -90 to -30 (mode -80)ndash Depolarization potential 20-60 mV ndash Duration of depolarization 50 ms ndash 10s (mode 2s)ndash Repolarization potential -80 to -10 mV (mode -40 mV)ndash Duration of repolarization 05 ndash 5s (mode 2s)
bull IKs measurement steady state current at the end of depolarization and tail currents
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation (s mV)
deactivation(s mV)
Interval (s)
47 hKvLQT1hminK CHO RT -80 2 40 01 Hz
49 hKv711hKCNE1 CHO RT -80 2 20 05 -40 15s
50 hKv711hKCNE1 HEk293 RT -80 2 20 05 -40 15s
184 IKs canineepiardial cells 37 -50 2 40 na -30 na
214 Kv711minK HEK293 na -80 5 -60 to 60 5s -50 na
215 IKs caninemyocyte na -40 5 50
224 hKCNQ1hKCNE1 HEK293 22-23 -80 3 -60 to 60 (20) na -50 na
229 hCNE1-hKCNQ1 Transgenic mousemyocyte RT -75 20 -40(Na block) 2 60 2 -40 006 Hz
230 KCNQ1KCNE1 XenopusOocyte 20-22 -80 5 30 na -70
232 KCNQ1KCNE1 HEK293 36 -80 2 -80 to 110 (10) 2 -60 6s
233 KCNQ1KCNE1 CHO na -80 80 -90 3 20 1 -50 005Hz
237 KCNQ1 CHO na -80 na -80 to 60 1 -40
239 KCNQ1KCNE1 HEK293 RT -70 1 -70 to 60 na -40
241 KCNQ1 andor minK MouseCOS-7 22-25 -80 3 -120 to 60 (20) 2 -30
242 hKvLQT1hminK HEK293 19-36 -80 3 -50 to 60 (10) 5 -50
242 hKvLQT1hminK HEK293 19-36 -50 1 or 2 50 na -50 10s
244 KCNQ1 andor KCNE1 CHO na -90 3 -70 to 70 (10) na -60
245 IKs GPMyocyte 35 -50 1 50
249 KCNQ1KCNE1 HEK293 33-34 -40 1 -40 to 80 (10) na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation(s mV)
Voltage dependent(s mV)
Deactivation(s mV)
Interval (s)
251 IKs GPMyocyte 22-2535-37 -40 2 60 2 -40 to 80 (20) na -40
252 IKs GPMyocyte 35 -80 200 -50 3 60 2 -40 60s
252 hminK XenopusOocyte 35 -80 10 20 10 -40 to 60 2 -80
256 IKs Rabbitmyocyte 34 -50 3 -40 to 50 na
260 hKCNQ1hKCNE1 HEK293 RT -80 25 -70 to 100 (10) 25 -40 15s
262 IKs GPMyocyte 35 -80 1 40 5 -10
263 Kv71 and minK CHO RT -80 2 40 na -40
264 IKs GPmyocyte na -80 200 -40 05 50 na -40
267 IKs GPmyocyte na -90 na 0 na 40 01- 015 0-80 -90
273 IKs Caninemyocyte 37 -40 3 50 na -40
274 IKs GPmyocyte 36 -30 05 50 or 2 70 05 (2) -30 to 70 na -30
275 KCNQ1KCNE1 CHO 21 -80 1 40 01Hz
276 hKCNQ1hminK CHO 22 -80 6 40 na -80 005Hz
278 IKs Canine myocyte 35-37 -50 3 -50 to 50 2s -40
283 KCNQ1KCNE1 HEK293 21-22 -80 3 40 3 -80 to 40 na -40
286 IKs GPmyocyte 34-36 -50 005 30 005 ndash3 -50 to 30 na -50
287 Iks GPmyocyte 36 -80 na -40 2 60 2 -40 to 80 na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
47 49 50
137 4 18 1 10 10 74 NaOH
184 145 4 18 18 10 11 NaH2PO4 1 E4031 (5uM) Cadmium (300 uM) 74 NaOH
214 140 4 2 1 10 74 NaOH
215 144 5 56 25 5 Dextrose 11 Nifedipine (5 uM) E4031 (1 uM) 74 NaOH
224 140 54 18 1 10 10 NaH2PO4 033 73 NaOH
229 132 48 2 12 10 5 E4031 nisoldipine 74 NaOH
230 96 2 18 1 5 74 NaOH
232 140 54 18 1 5 55 74 NaOH
233 17 120 4 2 1 10 10 74 NaOH
237 130 5 18 1 10 Na-Acetate 28 74 NaOH
239 150 54 18 4 10 74 NaOH
241 135 5 25 12 5 10 74 NaOH
242 132 4 18 12 10 111 74 NaOH
244 140 4 18 12 55 11 74 NaOH
249 140 54 18 05 5 55 NaH2PO4 033 74 NaOH
250 140 54 18 1 5 10 74 NaOH
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
251 1 1 2 10 N-methly-d-glucamine 132
lanthanum chloride 005 mMcadmium chloride 02 mM 74 HCl
252 140 47 13 1 10 10 74 NaOH
256 09 5 N-methly-d-glucamine 149
Nisoldipine 3 uME4031 10 uM 74 HCl
260 140 54 18 1 5 10 NaH2PO4 03 73 NaOH
262 137 4 18 2 10 115 Pyrubic acid 5 Thiamine 01 74 NaOH
263 130 4 1 1 10 10 TTX 003 mM CoCl2 2 mM 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10 KH2PO4 12 MgSO4 12 NaHCO3 44
Nifetidine 5 uM 4-aminopyridine 3 mM 72 NaOH
274 140 1 5 10 Cd2+ 02 mM 74 NaOH
275 130 54 18 1 10 5 74 NaOH
278 136 54 1 1 5 Dextrose 10 NaH2PO4 033 E4031 5 uM 74 NaOH
283 140 5 1 1 5 10 NaH2PO4 033 74 NaOH
286 09 5 5 N-methly-d-glucamine 149
Nisoldipine 04 uME4031 5 uM 74 NaOH
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
49 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
214 110 142 10 4 4 517 72 KOH
215 45 100 1 10 na 5 72
224 20 110 1 5 5 01 10 5 72 KOH
229 110 11 5 10 55 73
230 3000
232 20 125 10 10
233 140 1 1 20 73 KOH
237 10 125 1 10 5 10 10 72 KOH
239 30 130 01 4 10 74
241 117 2 11 11 10 1 72
242 15 119 32 5 5 5 74 KOH
244 130 1 5 5 10 74 KOH
249 130 5 5 01 5 5 74 HCl
250 80 40 10 5 01 5 40 065 72 HCl
251 50 60 1 11 5 10 1 74 KOH
252 140 11 1 10 10 72 KOH
amphotericin-B
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
256 80 40 10 5 5 5 065 72
262 150 1 5 4 5 5 73 KOH
263^ 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
274 40 106 1 5 4 5 72 KOH
276 20 110 1 10 5 10 10 1 72 KOH
278 20 110 1 10 5 01 10 5 72 KOH
283 20 110 1 5 5 01 1 5 72 KOH
286 50 1 5 5 01 5 2 10 72 KOH
^phosphatidylinositol 45-diphosphate (5 mgml 1) was added to the internal solution to avoid the rundown of the current 70 L-aspartic acid 70 KOH
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation (s mV)
deactivation(s mV)
Interval (s)
47 hKvLQT1hminK CHO RT -80 2 40 01 Hz
49 hKv711hKCNE1 CHO RT -80 2 20 05 -40 15s
50 hKv711hKCNE1 HEk293 RT -80 2 20 05 -40 15s
184 IKs canineepiardial cells 37 -50 2 40 na -30 na
214 Kv711minK HEK293 na -80 5 -60 to 60 5s -50 na
215 IKs caninemyocyte na -40 5 50
224 hKCNQ1hKCNE1 HEK293 22-23 -80 3 -60 to 60 (20) na -50 na
229 hCNE1-hKCNQ1 Transgenic mousemyocyte RT -75 20 -40(Na block) 2 60 2 -40 006 Hz
230 KCNQ1KCNE1 XenopusOocyte 20-22 -80 5 30 na -70
232 KCNQ1KCNE1 HEK293 36 -80 2 -80 to 110 (10) 2 -60 6s
233 KCNQ1KCNE1 CHO na -80 80 -90 3 20 1 -50 005Hz
237 KCNQ1 CHO na -80 na -80 to 60 1 -40
239 KCNQ1KCNE1 HEK293 RT -70 1 -70 to 60 na -40
241 KCNQ1 andor minK MouseCOS-7 22-25 -80 3 -120 to 60 (20) 2 -30
242 hKvLQT1hminK HEK293 19-36 -80 3 -50 to 60 (10) 5 -50
242 hKvLQT1hminK HEK293 19-36 -50 1 or 2 50 na -50 10s
244 KCNQ1 andor KCNE1 CHO na -90 3 -70 to 70 (10) na -60
245 IKs GPMyocyte 35 -50 1 50
249 KCNQ1KCNE1 HEK293 33-34 -40 1 -40 to 80 (10) na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation(s mV)
Voltage dependent(s mV)
Deactivation(s mV)
Interval (s)
251 IKs GPMyocyte 22-2535-37 -40 2 60 2 -40 to 80 (20) na -40
252 IKs GPMyocyte 35 -80 200 -50 3 60 2 -40 60s
252 hminK XenopusOocyte 35 -80 10 20 10 -40 to 60 2 -80
256 IKs Rabbitmyocyte 34 -50 3 -40 to 50 na
260 hKCNQ1hKCNE1 HEK293 RT -80 25 -70 to 100 (10) 25 -40 15s
262 IKs GPMyocyte 35 -80 1 40 5 -10
263 Kv71 and minK CHO RT -80 2 40 na -40
264 IKs GPmyocyte na -80 200 -40 05 50 na -40
267 IKs GPmyocyte na -90 na 0 na 40 01- 015 0-80 -90
273 IKs Caninemyocyte 37 -40 3 50 na -40
274 IKs GPmyocyte 36 -30 05 50 or 2 70 05 (2) -30 to 70 na -30
275 KCNQ1KCNE1 CHO 21 -80 1 40 01Hz
276 hKCNQ1hminK CHO 22 -80 6 40 na -80 005Hz
278 IKs Canine myocyte 35-37 -50 3 -50 to 50 2s -40
283 KCNQ1KCNE1 HEK293 21-22 -80 3 40 3 -80 to 40 na -40
286 IKs GPmyocyte 34-36 -50 005 30 005 ndash3 -50 to 30 na -50
287 Iks GPmyocyte 36 -80 na -40 2 60 2 -40 to 80 na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
47 49 50
137 4 18 1 10 10 74 NaOH
184 145 4 18 18 10 11 NaH2PO4 1 E4031 (5uM) Cadmium (300 uM) 74 NaOH
214 140 4 2 1 10 74 NaOH
215 144 5 56 25 5 Dextrose 11 Nifedipine (5 uM) E4031 (1 uM) 74 NaOH
224 140 54 18 1 10 10 NaH2PO4 033 73 NaOH
229 132 48 2 12 10 5 E4031 nisoldipine 74 NaOH
230 96 2 18 1 5 74 NaOH
232 140 54 18 1 5 55 74 NaOH
233 17 120 4 2 1 10 10 74 NaOH
237 130 5 18 1 10 Na-Acetate 28 74 NaOH
239 150 54 18 4 10 74 NaOH
241 135 5 25 12 5 10 74 NaOH
242 132 4 18 12 10 111 74 NaOH
244 140 4 18 12 55 11 74 NaOH
249 140 54 18 05 5 55 NaH2PO4 033 74 NaOH
250 140 54 18 1 5 10 74 NaOH
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
251 1 1 2 10 N-methly-d-glucamine 132
lanthanum chloride 005 mMcadmium chloride 02 mM 74 HCl
252 140 47 13 1 10 10 74 NaOH
256 09 5 N-methly-d-glucamine 149
Nisoldipine 3 uME4031 10 uM 74 HCl
260 140 54 18 1 5 10 NaH2PO4 03 73 NaOH
262 137 4 18 2 10 115 Pyrubic acid 5 Thiamine 01 74 NaOH
263 130 4 1 1 10 10 TTX 003 mM CoCl2 2 mM 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10 KH2PO4 12 MgSO4 12 NaHCO3 44
Nifetidine 5 uM 4-aminopyridine 3 mM 72 NaOH
274 140 1 5 10 Cd2+ 02 mM 74 NaOH
275 130 54 18 1 10 5 74 NaOH
278 136 54 1 1 5 Dextrose 10 NaH2PO4 033 E4031 5 uM 74 NaOH
283 140 5 1 1 5 10 NaH2PO4 033 74 NaOH
286 09 5 5 N-methly-d-glucamine 149
Nisoldipine 04 uME4031 5 uM 74 NaOH
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
49 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
214 110 142 10 4 4 517 72 KOH
215 45 100 1 10 na 5 72
224 20 110 1 5 5 01 10 5 72 KOH
229 110 11 5 10 55 73
230 3000
232 20 125 10 10
233 140 1 1 20 73 KOH
237 10 125 1 10 5 10 10 72 KOH
239 30 130 01 4 10 74
241 117 2 11 11 10 1 72
242 15 119 32 5 5 5 74 KOH
244 130 1 5 5 10 74 KOH
249 130 5 5 01 5 5 74 HCl
250 80 40 10 5 01 5 40 065 72 HCl
251 50 60 1 11 5 10 1 74 KOH
252 140 11 1 10 10 72 KOH
amphotericin-B
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
256 80 40 10 5 5 5 065 72
262 150 1 5 4 5 5 73 KOH
263^ 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
274 40 106 1 5 4 5 72 KOH
276 20 110 1 10 5 10 10 1 72 KOH
278 20 110 1 10 5 01 10 5 72 KOH
283 20 110 1 5 5 01 1 5 72 KOH
286 50 1 5 5 01 5 2 10 72 KOH
^phosphatidylinositol 45-diphosphate (5 mgml 1) was added to the internal solution to avoid the rundown of the current 70 L-aspartic acid 70 KOH
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Pulse sequences IKsID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse (ms mV)
Activation(s mV)
Voltage dependent(s mV)
Deactivation(s mV)
Interval (s)
251 IKs GPMyocyte 22-2535-37 -40 2 60 2 -40 to 80 (20) na -40
252 IKs GPMyocyte 35 -80 200 -50 3 60 2 -40 60s
252 hminK XenopusOocyte 35 -80 10 20 10 -40 to 60 2 -80
256 IKs Rabbitmyocyte 34 -50 3 -40 to 50 na
260 hKCNQ1hKCNE1 HEK293 RT -80 25 -70 to 100 (10) 25 -40 15s
262 IKs GPMyocyte 35 -80 1 40 5 -10
263 Kv71 and minK CHO RT -80 2 40 na -40
264 IKs GPmyocyte na -80 200 -40 05 50 na -40
267 IKs GPmyocyte na -90 na 0 na 40 01- 015 0-80 -90
273 IKs Caninemyocyte 37 -40 3 50 na -40
274 IKs GPmyocyte 36 -30 05 50 or 2 70 05 (2) -30 to 70 na -30
275 KCNQ1KCNE1 CHO 21 -80 1 40 01Hz
276 hKCNQ1hminK CHO 22 -80 6 40 na -80 005Hz
278 IKs Canine myocyte 35-37 -50 3 -50 to 50 2s -40
283 KCNQ1KCNE1 HEK293 21-22 -80 3 40 3 -80 to 40 na -40
286 IKs GPmyocyte 34-36 -50 005 30 005 ndash3 -50 to 30 na -50
287 Iks GPmyocyte 36 -80 na -40 2 60 2 -40 to 80 na -40
Iks (Kv71minK kvLQT1 KCNQ1KCNE1)
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
47 49 50
137 4 18 1 10 10 74 NaOH
184 145 4 18 18 10 11 NaH2PO4 1 E4031 (5uM) Cadmium (300 uM) 74 NaOH
214 140 4 2 1 10 74 NaOH
215 144 5 56 25 5 Dextrose 11 Nifedipine (5 uM) E4031 (1 uM) 74 NaOH
224 140 54 18 1 10 10 NaH2PO4 033 73 NaOH
229 132 48 2 12 10 5 E4031 nisoldipine 74 NaOH
230 96 2 18 1 5 74 NaOH
232 140 54 18 1 5 55 74 NaOH
233 17 120 4 2 1 10 10 74 NaOH
237 130 5 18 1 10 Na-Acetate 28 74 NaOH
239 150 54 18 4 10 74 NaOH
241 135 5 25 12 5 10 74 NaOH
242 132 4 18 12 10 111 74 NaOH
244 140 4 18 12 55 11 74 NaOH
249 140 54 18 05 5 55 NaH2PO4 033 74 NaOH
250 140 54 18 1 5 10 74 NaOH
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
251 1 1 2 10 N-methly-d-glucamine 132
lanthanum chloride 005 mMcadmium chloride 02 mM 74 HCl
252 140 47 13 1 10 10 74 NaOH
256 09 5 N-methly-d-glucamine 149
Nisoldipine 3 uME4031 10 uM 74 HCl
260 140 54 18 1 5 10 NaH2PO4 03 73 NaOH
262 137 4 18 2 10 115 Pyrubic acid 5 Thiamine 01 74 NaOH
263 130 4 1 1 10 10 TTX 003 mM CoCl2 2 mM 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10 KH2PO4 12 MgSO4 12 NaHCO3 44
Nifetidine 5 uM 4-aminopyridine 3 mM 72 NaOH
274 140 1 5 10 Cd2+ 02 mM 74 NaOH
275 130 54 18 1 10 5 74 NaOH
278 136 54 1 1 5 Dextrose 10 NaH2PO4 033 E4031 5 uM 74 NaOH
283 140 5 1 1 5 10 NaH2PO4 033 74 NaOH
286 09 5 5 N-methly-d-glucamine 149
Nisoldipine 04 uME4031 5 uM 74 NaOH
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
49 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
214 110 142 10 4 4 517 72 KOH
215 45 100 1 10 na 5 72
224 20 110 1 5 5 01 10 5 72 KOH
229 110 11 5 10 55 73
230 3000
232 20 125 10 10
233 140 1 1 20 73 KOH
237 10 125 1 10 5 10 10 72 KOH
239 30 130 01 4 10 74
241 117 2 11 11 10 1 72
242 15 119 32 5 5 5 74 KOH
244 130 1 5 5 10 74 KOH
249 130 5 5 01 5 5 74 HCl
250 80 40 10 5 01 5 40 065 72 HCl
251 50 60 1 11 5 10 1 74 KOH
252 140 11 1 10 10 72 KOH
amphotericin-B
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
256 80 40 10 5 5 5 065 72
262 150 1 5 4 5 5 73 KOH
263^ 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
274 40 106 1 5 4 5 72 KOH
276 20 110 1 10 5 10 10 1 72 KOH
278 20 110 1 10 5 01 10 5 72 KOH
283 20 110 1 5 5 01 1 5 72 KOH
286 50 1 5 5 01 5 2 10 72 KOH
^phosphatidylinositol 45-diphosphate (5 mgml 1) was added to the internal solution to avoid the rundown of the current 70 L-aspartic acid 70 KOH
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
47 49 50
137 4 18 1 10 10 74 NaOH
184 145 4 18 18 10 11 NaH2PO4 1 E4031 (5uM) Cadmium (300 uM) 74 NaOH
214 140 4 2 1 10 74 NaOH
215 144 5 56 25 5 Dextrose 11 Nifedipine (5 uM) E4031 (1 uM) 74 NaOH
224 140 54 18 1 10 10 NaH2PO4 033 73 NaOH
229 132 48 2 12 10 5 E4031 nisoldipine 74 NaOH
230 96 2 18 1 5 74 NaOH
232 140 54 18 1 5 55 74 NaOH
233 17 120 4 2 1 10 10 74 NaOH
237 130 5 18 1 10 Na-Acetate 28 74 NaOH
239 150 54 18 4 10 74 NaOH
241 135 5 25 12 5 10 74 NaOH
242 132 4 18 12 10 111 74 NaOH
244 140 4 18 12 55 11 74 NaOH
249 140 54 18 05 5 55 NaH2PO4 033 74 NaOH
250 140 54 18 1 5 10 74 NaOH
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
251 1 1 2 10 N-methly-d-glucamine 132
lanthanum chloride 005 mMcadmium chloride 02 mM 74 HCl
252 140 47 13 1 10 10 74 NaOH
256 09 5 N-methly-d-glucamine 149
Nisoldipine 3 uME4031 10 uM 74 HCl
260 140 54 18 1 5 10 NaH2PO4 03 73 NaOH
262 137 4 18 2 10 115 Pyrubic acid 5 Thiamine 01 74 NaOH
263 130 4 1 1 10 10 TTX 003 mM CoCl2 2 mM 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10 KH2PO4 12 MgSO4 12 NaHCO3 44
Nifetidine 5 uM 4-aminopyridine 3 mM 72 NaOH
274 140 1 5 10 Cd2+ 02 mM 74 NaOH
275 130 54 18 1 10 5 74 NaOH
278 136 54 1 1 5 Dextrose 10 NaH2PO4 033 E4031 5 uM 74 NaOH
283 140 5 1 1 5 10 NaH2PO4 033 74 NaOH
286 09 5 5 N-methly-d-glucamine 149
Nisoldipine 04 uME4031 5 uM 74 NaOH
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
49 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
214 110 142 10 4 4 517 72 KOH
215 45 100 1 10 na 5 72
224 20 110 1 5 5 01 10 5 72 KOH
229 110 11 5 10 55 73
230 3000
232 20 125 10 10
233 140 1 1 20 73 KOH
237 10 125 1 10 5 10 10 72 KOH
239 30 130 01 4 10 74
241 117 2 11 11 10 1 72
242 15 119 32 5 5 5 74 KOH
244 130 1 5 5 10 74 KOH
249 130 5 5 01 5 5 74 HCl
250 80 40 10 5 01 5 40 065 72 HCl
251 50 60 1 11 5 10 1 74 KOH
252 140 11 1 10 10 72 KOH
amphotericin-B
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
256 80 40 10 5 5 5 065 72
262 150 1 5 4 5 5 73 KOH
263^ 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
274 40 106 1 5 4 5 72 KOH
276 20 110 1 10 5 10 10 1 72 KOH
278 20 110 1 10 5 01 10 5 72 KOH
283 20 110 1 5 5 01 1 5 72 KOH
286 50 1 5 5 01 5 2 10 72 KOH
^phosphatidylinositol 45-diphosphate (5 mgml 1) was added to the internal solution to avoid the rundown of the current 70 L-aspartic acid 70 KOH
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Extracellular buffer solution of IKsID NaCl Na-
Gluc KCl CaCl2 MgCl2 HEPES glucose Minor components Blocker pH pH buffer
251 1 1 2 10 N-methly-d-glucamine 132
lanthanum chloride 005 mMcadmium chloride 02 mM 74 HCl
252 140 47 13 1 10 10 74 NaOH
256 09 5 N-methly-d-glucamine 149
Nisoldipine 3 uME4031 10 uM 74 HCl
260 140 54 18 1 5 10 NaH2PO4 03 73 NaOH
262 137 4 18 2 10 115 Pyrubic acid 5 Thiamine 01 74 NaOH
263 130 4 1 1 10 10 TTX 003 mM CoCl2 2 mM 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10 KH2PO4 12 MgSO4 12 NaHCO3 44
Nifetidine 5 uM 4-aminopyridine 3 mM 72 NaOH
274 140 1 5 10 Cd2+ 02 mM 74 NaOH
275 130 54 18 1 10 5 74 NaOH
278 136 54 1 1 5 Dextrose 10 NaH2PO4 033 E4031 5 uM 74 NaOH
283 140 5 1 1 5 10 NaH2PO4 033 74 NaOH
286 09 5 5 N-methly-d-glucamine 149
Nisoldipine 04 uME4031 5 uM 74 NaOH
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
49 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
214 110 142 10 4 4 517 72 KOH
215 45 100 1 10 na 5 72
224 20 110 1 5 5 01 10 5 72 KOH
229 110 11 5 10 55 73
230 3000
232 20 125 10 10
233 140 1 1 20 73 KOH
237 10 125 1 10 5 10 10 72 KOH
239 30 130 01 4 10 74
241 117 2 11 11 10 1 72
242 15 119 32 5 5 5 74 KOH
244 130 1 5 5 10 74 KOH
249 130 5 5 01 5 5 74 HCl
250 80 40 10 5 01 5 40 065 72 HCl
251 50 60 1 11 5 10 1 74 KOH
252 140 11 1 10 10 72 KOH
amphotericin-B
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
256 80 40 10 5 5 5 065 72
262 150 1 5 4 5 5 73 KOH
263^ 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
274 40 106 1 5 4 5 72 KOH
276 20 110 1 10 5 10 10 1 72 KOH
278 20 110 1 10 5 01 10 5 72 KOH
283 20 110 1 5 5 01 1 5 72 KOH
286 50 1 5 5 01 5 2 10 72 KOH
^phosphatidylinositol 45-diphosphate (5 mgml 1) was added to the internal solution to avoid the rundown of the current 70 L-aspartic acid 70 KOH
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
49 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
214 110 142 10 4 4 517 72 KOH
215 45 100 1 10 na 5 72
224 20 110 1 5 5 01 10 5 72 KOH
229 110 11 5 10 55 73
230 3000
232 20 125 10 10
233 140 1 1 20 73 KOH
237 10 125 1 10 5 10 10 72 KOH
239 30 130 01 4 10 74
241 117 2 11 11 10 1 72
242 15 119 32 5 5 5 74 KOH
244 130 1 5 5 10 74 KOH
249 130 5 5 01 5 5 74 HCl
250 80 40 10 5 01 5 40 065 72 HCl
251 50 60 1 11 5 10 1 74 KOH
252 140 11 1 10 10 72 KOH
amphotericin-B
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
256 80 40 10 5 5 5 065 72
262 150 1 5 4 5 5 73 KOH
263^ 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
274 40 106 1 5 4 5 72 KOH
276 20 110 1 10 5 10 10 1 72 KOH
278 20 110 1 10 5 01 10 5 72 KOH
283 20 110 1 5 5 01 1 5 72 KOH
286 50 1 5 5 01 5 2 10 72 KOH
^phosphatidylinositol 45-diphosphate (5 mgml 1) was added to the internal solution to avoid the rundown of the current 70 L-aspartic acid 70 KOH
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Pipette solution of IKsID KCl K-asp
K-glucona
te
K-glutama
teMgCl2 EGTA ATP GTP HEPES Na2-
PhosCre NaCl CaCl2 KH2PO4 pH pH buffer
256 80 40 10 5 5 5 065 72
262 150 1 5 4 5 5 73 KOH
263^ 42 80 5 5 5 3 10 72 KOH
267 135 5 5 5-10 10 19 72 KOH
273 45 100 1 10 3 5 72 KOH
274 40 106 1 5 4 5 72 KOH
276 20 110 1 10 5 10 10 1 72 KOH
278 20 110 1 10 5 01 10 5 72 KOH
283 20 110 1 5 5 01 1 5 72 KOH
286 50 1 5 5 01 5 2 10 72 KOH
^phosphatidylinositol 45-diphosphate (5 mgml 1) was added to the internal solution to avoid the rundown of the current 70 L-aspartic acid 70 KOH
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Summary of IK1 protocolsbull Distribution higher in ventricle than atrial
bull Pharmacology establish resting membrane potential selective antiarrhythmia strategy
bull Channel sources1 Transfected cell lines CHO HEK 293 and Xenopus Oocyte (831 = 26)2 Myocyte (74) species ndash Canine VM rat VM Frog VM Fish VM Cat VM Guinea pig VM
and rabbit VM (mode Rat and GP)
bull Temperaturendash RT = 1020 50 ndash physiological temperature = 50
bull Pulse sequencesndash Holding range = -120 to -30 (mode -40)ndash Activation range -140 to 100 mV (mostly voltage dependence protocols)ndash Duration of activation 01 to 4s (mode 05s)
bull IK1 measurement steady state current at the end of depolarization
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Pulse sequences IK1
ID ChannelCurrent
Speciescells Temp Vh(mV)
Potency (s mV)
Voltage -Dependence(m mV)
Interval Frequency
47 hKir21 HamsterCHO RT -30 1 -120 01 Hz
184 IK1 CanineMyocyte 37 -40 1 -100 to 0
206 IK1 RatMyocyte 24 -40 05 -120 to 20
207 IK1 RatMyocyte 35 -40 05 -10 to 100 (10) 5s
208 IK1 RatMyocyte 23 -40 03 -120 to 50 3s
209 IK1 FrogMyocyte 20-21 -100 na -120 to 50 02 Hz
210 IK1 HumanAM 32-34 -75 05 -100
213 IK1 FishMyocyte 18 -80 1 -120 to 30 5s
213 IK1 RatMyocyte 375 -40 1 -120 to 30 5s
214 hKIr21 22 23 HumanHEK293 na -40 na -100 to 100 5s
215 IK1 CanineMyocyte na -40 04 -135 to -65
216 IK1 RatMyocyte 20-22 -80 na -120 to -80
217 Kir21 23 HumanHEK293 na -80 4 -120 to -20
217 IK1 CatMyocyte na -40 3 -120 to -30
220 IK1 RatMyocyte RT -120 na -135 to -75
222 IK1 RatMyocyte 22-24 -80 4 -120 to 50
224 hKIr21 HumanHEK293 22-23 -40 na -120 to 0
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Pulse sequences IK1ID Channel
CurrentSpeciescells Temp Vh
(mV)Prepulse(ms mV)
Potency (ms mV)
Voltage -Dependence(ms mV)
IntervalFrequency
225 Kir21 HEK293 na -40 na -117 na -120 to 0
225 Kv21 CHO -70 na -70 to na
226 Kir21 HEK293 na -80 4 -140 to 0 (10)
245 IK1 GPMyocyte 35 -50 05 -80 to- 50 (ramp)
247 IK1 GPMyocyte na -40 025 -110 to 60 (10)
252 Ik1 GPMyocyte 35 -80 02 -120
256 IK1 RabbitMyocyte 34 -40 03 -100 to 50 01 Hz
262 IK1 GPMyocyte 35 na 01 -100
264 IK1 GPMyocyte -40-80 05 -40 05 -120 to 0
264 IK1 GPMyocyte -80 05 -40
267 IK1 GPMyocyte -40 na -140 to 0
273 IK1 CanineM 37 -80 04 -125 to 65
274 IK1 GPMyocyte 36 -80 -90 -40 05 -40 05 -120 to -10
281 Kir21 XenopusOocyte 22 -40 05 -120
287 IK1 GPMyocyte 36 -80 05 -40 01 Hz
-80 mV
-40 mV
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Sepcial
components Blocker (mM) pH pH buffer
47 137 4 18 1 10 10 74 NaOH
184 132 4 2 12 20 11 Ouabain (3uM) nifedipine (5uM) 74 NaOH
206 135 54 18 1 10 10 NaH2PO4 (033) 74 NaOH
207 140 54 18 15 10 55 NaH2PO4 (033) 10 uM Nifedipine BaCl2 (1mM) 74 NaOH
208 136 54 18 1 10 10 Cd2+ (03) 4-AP (2) 74 NaOH
209 1105 25 2 1 10 10 Na-Pyrubate (5)CdCl2 (1) TTA (00006) 4-AP (2) TEA (10) CoCl2 (1) NiCl2 (05) SITS (2)
74 NaOH
210 137 4 18 12 10 dextrose (11) CdCl2 (02) 74 NaOH
213 150 54 18 12 10 10 TTX (05 uM) nifedipine (001) E4031 (0001) 76 NaOH
214-1 125 4 EDTA (4) KH2PO4 (28) K2HPO4 (72) 72 NaOH
214-2 140 5 18 1 6 6
215 144 56 25 12 5 Dextrose (11) Nifedipine ( 0005) E4031 (0001) 74 na
216 140 5 1 1 10 10 CdCl2 (02) 74 na
217 130 4 18 1 10 10 74 NaOH
217 136 54 01 1 10 11 CoCl2 (05) 74 na
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Extracellular buffer solution of IK1ID NaCl KCl CaCl2 MgCl2 HEPES glucose Special comp Blocker (mM) pH pH buffer
220 135 54 09 09 10 10 NaH2Po4 (033) CoCl2 (2) 4-AP (3) 74 NaOH
222 135 4 18 1 2 11 TTX (001) Co2+ (2) 74 NaOH
224 140 54 18 1 10 10 NaH2Po4 (033) 73 NaOH
225 140 54 2 08 10 10 74 NaOH
226 130 4 18 1 10 10 74 NaOH
247 140 54 05 5 55 NaH2Po4 (033) Nisoldipine (04) 74 NaOH
252 140 47 13 1 10 10 74 NaOH
262 137 4 18 2 10 115 Pyruvic acid (5) thiamine (01) 74 NaOH
264 140 54 1 5 10 74 NaOH
267 137 54 2 1 10 10 74 NaOH
273 135 47 10 10KH2PO4 (12) MgSO4 (12) NaHCO3 (44)
Nifedipine (5) 4-aminopyridine (3) 74 NaOH
281 96 2 18 1 5 73 NaOH
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Pipette solution of IK1ID KCl K-aspartate K-glutamate MgCl2 EGTA ATP GTP HEPES Special component pH pH
buffer
47 130 5 5 4 10 72 KOH
184 20 125 1 10 5 5 71 KOH
206 150 1 5 3 5 74 KOH
207 10 130 1 10 CaCl2 (05) glucose (10) Phosphocreatinine (10) 72 KOH
208 20 100 1 10 5 5 KOH (110) 72 KOH
209 150 5 5 10 Phosphocreatinine (5) 723 KOH
210 130 1 5 5 5 NaCl (7) 72 KOH
213 140 1 5 4 003 10 72 KOH
214 145 5 CaCl2 (1) 72 KOH
215 45 100 1 10 Not indicated 5 72
216 20 130 1 5 10 NaCl (5) 74
217-1 110 1 5 10 K4BAPTA (5) 72
217-2 123 5 K2HPO4 (72) KH2PO4 (8) Na3VO4 (01) KF (5) K4P2O7 (10) 72
217-3 40 80 10 3 02 5 KH2PO4 (10) MgSo4 (1) phosphocreatinine (5) 735
220 25 130 1 1 5 01 5 Phosphocreatinine (3) 725 KOH
222 130 05 3 04 25 72 KOH
224 20 110 1 5 5 01 10 Phosphocreatinine (5) 72 KOH
225 140 2 1 1 10 72 KOH
226 100 1 5 10 K4BAPTA (5) 72 KOH
245 25 500 5 72 KOH
247 30 70 3 5 5 01 5 KH2PO4 (10) 72 KOH
252 140 11 1 10 NaCl (10) 72 KOH
262 150 1 5 4 5 Hydroxybutyric acid (2) Phosphocreatine (5) 73 KOH
264 274 287 40 106 1 5 4 5 72 KOH
267 135 5 5 5-10 NaCl (10) CaCl2 (19) 72 KOH
273 45 100 1 10 3 5 72 KOH
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Summary of minor K protocols
summary 28
IKr Ito1 IKur IKs IK1
Sample literatures 18 18 8 36 31
cells cell line gt myocyte cell line gt myocyte cell line gt myocyte cell line myocyte cell line lt myocyte
Temp RT lt Phy RT gt Phy(80 vs 20)
RT(100)
RT Phy( 55 vs 45)
RT Phy (50 vs 50)
Pulses (mode) for IC50
Vh= -80 mV Vd= 0-70 mV Td=1-4s Vr= -80 -+20 Tr= 2-6s
Vh= -90 to -40 (-80)Vd= 0-70 mV (50) Td= 150- 500 ms (300)
Vh= -80 to -50 (-80)Vd=20-60 mV (50) Td= 150- 1000 ms (300)
Vh= -90 to -30 (-80)Vd=20-60 mV (40) Td= 05- 10s (2)Vr= -80 to -10 (-40)Tr= 05 ndash 5 s (2s )
Vh= -120 to -30 (-40)Vd=-140 to 100 mV (hyperpolarizing start from -120)Td=01 ndash 4s (05 s)
measure Tail current Peak current Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization and tail current
Steady state current at the end of depolarization
pulse trace
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Extracellular buffer IKr Ito Ikur Iks Ik1
NaCl
KCl
CaCl2
MgCl2
HEPES
glucose
Na2HPO4 () () () ()
Na-acetate ()
NMDG ()
Na-acetate () () ()
NMDG ()
dextrose () () () ()
K-gluconate () () ()
Na-pyruvate
EDTA ()
KH2PO4K2HPO4 () ()
Blockers () included in less than half of protocols summary 29
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Intracellular buffer IKr Ito Ikur Iks Ik1
Current carrier K-aspartateK-gluconateK-glutamate
KCl
MgCl2
EGTA
ATP Na2ATPK2ATPMgATP
HEPES
GTP () () () ()
ATPregeneration system phosphocreatinine () () () ()
CaCl2 () () () ()
NaCl () () () ()
KOH () ()
KH2PO4 () () ()
glucose ()
K4BAPTA ()
summary 30
() included in less than half of protocols
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
Representative current traces Ito (Kir 43)IKr
IK1 (Kir21)
Ikur (Kv15)
Iks (Kv71minK kvLQT1)
ICa
INa
ID 214 217
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
ReferencesID Journal Year volume first page47 J Cariovasc Pharmacol 2010 56 42049 J Cardiolvasc Electrophysiol 2010 21 30150 J Pharmacol Exp Ther 2008 324 427100 Am J Physiol Heart Circ Physiol 1998 275 H2016160 Front Pharmacol 2013 4 1173 J Clin Invest 1997 99 1714174 J Cariovasc Pharmacol 1998 31 800177 Mol Pharmacol 2005 57 101178 J Mol Cell Cardiol 2001 33 923179 British Journal of Pharmacology 2000 130 1753180 J Mol Cell Cardiol 2002 34 1029184 Circulation 2004 110 904186 Cardiovascular Research 2006 69 116195 J Pharmacol Exp Ther 2013 344 23199 J Mol Cell Cardiol 2012 53 593205 Heart Rhythm 2009 6 1625206 J Cariovasc Pharmacol 2012 59 37207 Mol Cell Biochem 2011 352 117208 Cell Physiol Biochem 2010 26 967209 J Nat Prod 2006 69 558210 European Journal of Pharmacology 2006 529 47213 Comparative Biochemistry and physiology 2013 158 181214 Cardiovascular Research 2013 99 203215 European Journal of Pharmacology 2013 710 10216 Med Sci Mont 2011 17 BR165217 J Cariovasc Pharmacol 2011 57 407220 Gen Physiol Biophys 2005 24 311221 J Cardiolvasc Electrophysiol 2005 16 1227222 Am J Physiol Heart Circ Physiol 2005 288 H778224 Plos One 2012 7 e51550
ID Journal Year volume first page225 J Biol Chem 2009 284 15432226 PNAS 2008 105 1364229 J Biol Chem 2009 284 9140230 Pflugers Arch 2013 epub232 Circ Arrhythm Electrophysiol 2013 epub233 Hum Exp Toxicol 2013 epub237 Can J Physiol Pharmacol 2010 88 1181239 J cardiolvasc Pharmacol 2010 55 145241 Cardiovascular Research 2009 82 430242 Am J Physiol Heart Circ Physiol 2008 295 H1867244 Circulation Research 2006 98 1055245 J Pharmacol Exp Ther 2006 317 1054247 J Cariovasc Pharmacol 2005 46 167249 J Cariovasc Pharmacol 2004 43 178251 Anesth Analog 2003 96 1308252 Naunyn Schmiedebergs Arch Pharmacol 2002 365 29256 European Journal of Pharmacology 1999 376 189260 British Journal of Pharmacology 2010 159 93262 J Cariovasc Pharmacol 2004 44 244263 British Journal of Pharmacology 2005 146 146264 British Journal of Pharmacology 2000 131 1809267 J Pharmacol Exp Ther 1997 281 233273 British Journal of Pharmacology 2002 136 330274 British Journal of Pharmacology 2003 140 863275 European Journal of Pharmacology 2006 532 44276 J Cardiolvasc Electrophysiol 2005 16 329278 British Journal of Pharmacology 2004 142 1300281 British Journal of Pharmacology 1997 120 968283 British Journal of Pharmacology 2006 147 289286 J Cardiolvasc Electrophysiol 2005 16 314287 European Journal of Pharmacology 2000 401 137
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