History of Bioelectrical Study and the Electrophysiology ...€¦ · Evidence-BasedComplementaryandAlternativeMedicine 7 Voltage (V) Voltage (V) 400 600 800 1000 1200 1400 1600 1800

Hindawi Publishing CorporationEvidence-Based Complementary and Alternative MedicineVolume 2013 Article ID 486823 14 pageshttpdxdoiorg1011552013486823

Review ArticleHistory of Bioelectrical Study and the Electrophysiology ofthe Primo Vascular System

Sang Hyun Park1 Eung Hwi Kim1 Ho Jong Chang1 Seung Zhoo Yoon2

Ji Woong Yoon3 Seong-Jin Cho4 and Yeon-Hee Ryu45

1 Biomedical Team KAIST Institute for Information Technology Convergence Korea Advanced Institute of Science and Technology(KAIST) Daejeon 305-701 Republic of Korea

2Department of Anesthesiology and Pain Medicine College of Medicine Korea University Seoul 136-705 Republic of Korea3 Impedance Imaging Research Center ampDepartment of Public Administration Kyung Hee University Seoul 130-701 Republic of Korea4Acupuncture Moxibustion amp Meridian Research Center Division of Standard Research Korea Institute of Oriental MedicineDaejeon 305-811 Republic of Korea

5Medical Research Division Acupuncture Moxibustion and Meridian Research Group Korea Institute of Oriental Medicine1672 Yuseongdae-ro Yuseong-gu Daejeon 305-811 Republic of Korea

Correspondence should be addressed to Yeon-Hee Ryu yhryukiomrekr

Received 22 February 2013 Revised 3 May 2013 Accepted 7 May 2013

Academic Editor Byung-Cheon Lee

Copyright copy 2013 Sang Hyun Park et alThis is an open access article distributed under theCreative CommonsAttribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Background Primo vascular system is a new anatomical structure whose research results have reported the possibility of a newcirculatory system similar to the blood vascular system and cells Electrophysiology which measures and analyzes bioelectricalsignals tissues and cells is an important research area for investigating the function of tissues and cells The bioelectrical study ofthe primo vascular system has been reported by usingmodern techniques since the early 1960s by BonghanKimThis paper reviewsthe research result of the electrophysiological study of the primo vascular system for the discussion of the circulatory function Wehope it would help to study the electrophysiology of the primo vascular system for researchers This paper will use the followingexchangeable expressions Kyungrak system = Bonghan system = Bonghan circulatory system = primo vascular system = primosystem Bonghan corpuscle = primo node Bonghan duct = primo vessel We think that objective descriptions of reviewed papersare more important than unified expressions when citing the papers That said this paper will unify the expressions of the primovascular system

1 Primo Vascular System

The primo vascular system has been studied as part of theanatomical and histological research of the last 10 years Itis a novel thread-like structure that remains unrevealed inanimals and humans Particularly the primo vascular systemwas selected as the feature article and for the cover page inthe Journal of Anatomical Record [1] it provoked controversyin the anatomical society and among acupuncture scientistsSince then the discovery of the primo vascular system inlymphatic vessels [2] cardiac vascular vessels [3] and thebrain [4] has received much attention

Early scientists of the primo vascular systemwho focusedon anatomical and histological studies hypothesized about

the relationship between oriental medicine and acupunctureand the meridian These findings are helping to integrate thetheories of conventional and alternative medicine [5]

Kim who initially studied the anatomical structures ofthe acupuncture and themeridian declared that ldquoThis projectdeveloped from our inherited oriental medicine which is ourancestorsrsquo creative and noble endeavor as approved at thethird Joseon Labor Party Congressrdquo in his first paper [6]

Moreover Soh considered that Bonghan corpuscles andducts were identical to acupuncture sites and meridians [5]stating his theory that the Bonghan circulatory system maybe an extension of acupuncture and meridians [7] It wasreported that DNA was in the primo vascular system [1]

2 Evidence-Based Complementary and Alternative Medicine

C CopperZ ZincN Nerve

C

N

Z

Figure 1 The electrical stimulation of a frog nerve It was foundthat the current flows through the two different metals coming intocontact with animal muscle

evidence showing that the primo vascular system may be aDNA circulatory system has been reported as well [4]

Electrophysiology is a science of the electrical propertiesof biological tissues and cells It involves measurements ofvoltage changes or electric current on a wide variety of scalesfrom single-ion channel proteins to whole organs such as theheart In this paper we introduce the history of electricalsignals and the electrophysiology of the primo vascularsystem

Section 2 introduces the understanding of electrophysi-ology and modern techniques to measure the bioelectricalsignal for general readers Section 3 describes the beginningof the bioelectrical study of acupuncturemeridian briefly andreviews the electrophysiological study of the primo vascularsystem past and present The last section arranges the resultof the reviewed papers and suggests the future work for theelectrophysiology of the primo vascular system

2 Development of Electrophysiology withElectrical Signal and Basic Medicine

21 Bioelectrical Signal Studies and Electrophysiology In 1791the Italian physician andphysicist LuigiGalvani first recordedthe phenomenon of electrical signals while dissecting a frogon a table where he had been conducting experiments withstatic electricity (Figure 1) Galvani coined the term animalelectricity to describe the phenomenon while contempo-raries labeled it galvanism Galvani and his contemporariesregarded muscle activation as resulting from an electricalfluid or substance in the nerves [8 9]

In the work to study cardiac electrophysiology in orderto gain a better understanding of bioelectricity cardiacelectrophysiology emerged as an important area to elucidateand diagnose the heart function and to treat heart disease

including arrhythmia through electrocardiograms (Figure 2)[10] The function of tissues and organs in humans is closelyrelated to analyses of the bioelectrical signals from thesefeatures

22 Development of a Measurement Method for BioelectricSignals Electrophysiological studies which started fromfrog muscle response research extended the fundamentalknowledge of nerve cells through squid axon membranepotential measurements using an electrode when Hodgkinand Huxley began this work in 1952 [11] Graham and Gerardsucceeded to create and develop an electrode with a finerdiameter of 2ndash5 um to measure the intracellular potential(Figure 3) [12] With the development of this measurementtechnique Neher and Sakmann reduced the signal-to-noiseratio using a seamless seal between the cell membrane andthe electrode establishing the foundation of the study of ionchannels in the cell membrane [13] Cell membrane potentialmeasurement techniques in electrophysiological research areoutlined later

221 Intracellular Recording (Voltage Clamp and CurrentClamp) An electrical signal representing cell activities isa result of the activity of the ion channels in the cellmembrane The cell activity is associated with whether thecell membrane ion channel is activating or not and is thenrelated to the electrical signals from the cells Related to thisthe voltage clamp technique was developed to measure theintracellular ion flow with the constant membrane potentialThis technique was useful in research on the mechanism ofbiological electrical signals generated by the ions such as K+or Na+ passing through the electrically opened and closedchannels [11ndash13]

In contrast the current clamp was used to measure thechanges of the voltage inside the cell by means of a constantcurrent this was useful to classify cell types by understandingthe action potential of the cells (Figure 4) [14 15]

222 Patch Clamp As an intracellular recording techniquewhich involves inserting an electrode into a cell directlythe patch clamp technique was created for the purpose ofseparating and analyzing of each ion channel existing onthe membrane of a cell Patch clamp recordings use as anelectrode a glass micropipette that has an open tip diameterof about one micrometer (1120583m) which is a size enclosinga membrane surface area or ldquopatchrdquo that often contains justone or few ion channel molecules This type of electrode isdistinct from the ldquosharp microelectroderdquo used to impale cellsin traditional intracellular recordings in that it is sealed ontothe surface of the cell membrane rather than inserted throughit Many researchers including Neher and Sakmann studiedion channels in various cells using the patch clamp technique(Figure 5) [16ndash18]

They observed that the electrophysiological characteris-tics in each major tissue cell especially the resting potentialshowed a different form (Table 1) This cell-size level tech-nique led to the development of electrophysiology and has

Evidence-Based Complementary and Alternative Medicine 3

0 100 200 300 400 500 600 700Time (ms)

Sinusnode

Atrialmuscle

A-Vnode

CommonbundleBundle

branchesPurkinjefibers

Ventricularmuscle

P

Q

R

S

TU

Figure 2 Electrophysiology of the heart

Cell

Voltageelectrode

Figure 3 Intracellular recording technique

producedmany eminent scientists and Nobel Award winners[19ndash22]

The development of the measurement technique suchas intracellular recording and patch clamp advanced theelectrophysiological research

3 Beginning of Electrophysiology of the PrimoVascular System

Although the primo vascular system has been studiedintensively over last 10 years there is no complete evi-dence of whether the primo vascular system is an exten-sion of acupuncture and meridians system or is identicalto acupuncture and meridians which are key concepts

in oriental medicine Oriental medicine scientists scarcelyaccept hypotheses pertaining to the anatomical structuresof acupuncture and meridians However in their work onthe primo vascular system Soh [7] and Lee et al [23]declared that they were inspired by Bonghan Kimrsquos theoryfrom the 1960s [6 24ndash27] Therefore the beginning of theelectrophysiology of the primo vascular system establisheda standard from the beginning of the measurement of theelectrical properties related to acupuncture and meridians

31 Historical Trends of the Bioelectrical Study of Acupunctureand Meridians The bioelectrical signal measurements ofacupuncture and meridians were attempts to determine theexistence of these structures These studies were done usinganimals and cadavers it was assumed that human study wasonly partially complete

Jeh found that acupuncture points had lower resistancethan the surrounding skin by measuring the skin resistance[28] Overhof verified that the resistance at acupuncturepoints was lower than that of nonacupuncture points [29]Later Ogata et al reported that acupuncture points had lowerresistance than non-acupuncture points located in the samemeridian [30] In particular Niboyetrsquos methods have led tothe development of ear acupuncture which has become oneof the acupuncture treatment therapies offered in EuropeConsidering this low-resistance feature Voll designed anacupuncture diagnostic apparatus using microcurrents [31]

Studies of the low-resistance properties of acupuncturepoints have been performed intermittently Recently Ahnet al took ultrasonic images of acupuncture points andproposed anatomically that a collagenous band in connec-tive tissues was related to the low-impedance property ofacupuncture points [32]


Action potential

Depolarization

Hyperpolarization

1 2 3Time (ms)

Mem

bran

e pot

entia

l (m

V)

Neuron action potential

Thre

shol

d ex

cita

tion

5 4 3 2 1

Stimulusapplied

minus90

minus80

minus70

minus60

minus50

minus40

minus30

minus20

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010

20

30

40

(a)

Time (ms)0 2 4 6 8 10

10

20

30

40

50

60

70

0

20

40

60Action potential

Skeletal muscle action potential

minus60

minus40

minus20V(m

V)

G(m

scm

2)

Na+ conduction

K+ conduction

(b)

Cardiac muscle action potential

50

0

minus50

minus100

(mV

)

4

0

12

3

4

(c)

Mem

bran

e pot

entia

l (m

V)

Time (ms)0 2 4 6

Smooth muscle action potential

(times104)

minus70

minus60

minus50

minus40

(d)

Figure 4 A comparison of the action potentials of major tissue cells

A modern system to be able to measure the resistance ofacupuncture point has been developed recently based on theresult of the lower resistance on the acupoint [33]

But there are still limitations to measure the special pointon the skin by classical theory and method since 1950s

32 Bioelectrical Study and Electrophysiology of the PrimoVascular System in the 1960s Kim published five papersin total about the existence of the anatomical structure ofmeridians at the Kyungrak Institute in the 1960s [6 24ndash27 34] Three out of the five papers described electricalsignal measurements and analyses of acupuncture points andmeridians His first paper investigated the electrical prop-erties of acupuncture points and meridians The followingdescription was extracted from his first paper given at the

conference of the Pyongyang Medical School on October 181961

ldquoThis project was started to develop and prop-agate oriental medicine which is our ancestorrsquoscreative and noble endeavor as approved at thethird Joseon Labor Party Congress We have setourselves an assignment to find the electrical prop-erties of meridians first and then the Kyungraksystem based on these propertiesrdquo [6]

Kim already knew about research reports on the electricalresistance of acupuncture points showing that it was lowerthan the areas around the spots He had attempted tomeasureand examine unseen singular points on the skin Whenwriting his paper he investigated the electrical properties ofacupuncture by measuring the resistance and voltage around


On-cellpatch

Inside-outpatch

Whole-cellclamp

Outside-outpatch

(a) (b)

(c) (d)

Figure 5 Patch clamp technique

the acupuncture points of rabbits The resistance value wasabout 20000ndash80000Ω when applying 100120583A and it waslower than the values around the point He indicated theproblem of the changing measurement value in terms of themeasurement time interval and number of measurementsHowever the locations of the low-resistance points werefixed the distribution of the locations coincided with theacupuncture points described in the Dongui Bogam whichis a physician Book of Traditional Medicine compiled by HeoJun in 1613 during the Joseon Dynasty of Korea [35]

It was surprising that he had found that the voltage valueof the acupuncture point changed consistently The voltagechange was a regular and rhythmical wave group with a waveperiod of 3ndash6 sec and an intensity level of 01mV He reportedthat 5ndash7 waves were detected continuously followed by aresting phase (Figure 6) He asserted that non-acupuncturepoints did not display this phenomenon (Figure 7)

He considered that the changing values of acupuncturepoints may be connected to physiological properties andtherefore ran an interrelationship experiment

It was an experiment to measure the interactional electri-cal signals between stimulation by a needle and large intestinemovements He reported some interaction between acupointST36and themovement of the large intestinewhenmeasuring

the electrical signals and stimulating the intestine (Figure 8)His interpretation of the result from his interaction

experiment was different from those of general acupuncturescientists at that time He suggested that acupuncture andinternal organs should be connected materially to each otherhypothesized that acupuncture points and meridians wouldbe anatomical structures in the human body and started toexplore anatomical tissues (Figure 9)

B H Kim advanced anatomical and histological researchand Kyungrak circulatory research by developing his firstpaper into his second published paper entitled On theAcupuncture Meridian System

Figure 6 The electric induction on a Nogung acupuncture point(PC8)

Figure 7The induced electricity at non-acupuncture points located1 cm from Nogung

Table 1 A comparison of electrophysiological characteristicsbetween tissues

Neuron MuscleSkeletal Cardiac Smooth

Resting potential(mV) minus70 minus80 to minus90 minus85 to minus95 minus50 to minus60

Thresholdpotential (mV) minus55 minus55 Spike

potentialSpike

potentialAction potentialduration (ms) 1 2 to 5 200 to 400 10 to 50

He named the discovered anatomical structures aroundacupuncture points Bonghan corpuscles and the thread-likestructure connected to Bonghan corpuscles Bonghan ducts(He did not use the word Bonghan system He only namedthe new anatomical and histological structures of Kyungraksystem Bonghan corpuscles and Bonghan ducts [24])

In order to investigate the circulation of the Kyungraksystem he analyzed the physiological and bioelectrical mea-surements of Bonghan corpuscles first as this was the basisof the contention that the morphological characteristics ofBonghan corpuscles were smoothmuscles and secreting cells

He inserted an electrode directly into a Bonghan cor-puscle and measured the bioelectrical signal from Bonghancorpuscle It was very similar to the extracelluar recordingmethod in the modern technical term But there werelimitations to be able to measure the bioelectrical signalfrom the tissue of Bonghan corpuscle because of noises Nowmodern technique to measure the bioelectrical signal canmeasure the electrical signal from single cells [36]

The physiological study of Bonghan corpuscles and ductsprogressed as the manner in which bioelectrical signalschanged by various stimulations was studied

He proposed that Bonghan corpuscles and ductsresponded to various stimuli and that the responses weresimilar to those of nerves There were many experimentsconducted to determine where the signal was conductedmdashwhether it was along the Bonghan corpuscle and duct oranother area [6]

The bioelectrical research into Bonghan corpusclesrecorded the changing values of the bioelectrical signalsby inserting an electrode into a Bonghan corpuscle Theexperimental result reported that the change of the potential


Figure 8 The induced electricity at an acupuncture point afterhyperkinesis of the large intestine

Figure 9The electric induction on the Susamni acupuncture point(LI10)

Figure 10 The bioelectrical activity from a Bonghan corpuscle

Figure 11 The changes in the bioelectrical activity of a Bonghancorpuscle after an injection of acetylcholine

in a Bonghan corpuscle was similar to a sine curve termingthis curve a ldquoErdquo (geu) wave with a period of 3ndash6 secondsAlso identified was a ldquoFrdquo (neu) wave at 7ndash10 seconds anda ldquoGrdquo (deu) wave at 20ndash25 seconds The amplitude of eachwave was 01mV (Figure 10)

Particularly these electric potential changes disappearedwhen the temperature of the environment dropped below27∘Cbut reappearedwhen the temperaturewas raised to 39∘Cimmediately He suggested that the bioelectrical propertiesof acupuncture points as explained by the nervous systemwere incorrect as the phenomena of the changing signalappeared from Bonghan corpuscles which are not related tothe nervous system

The next experiment involved assessing the excitabilityand response of Bonghan corpuscle by various stimuli BH Kim expressed that ldquothe study of excitability of Bonghancorpuscles was a basic problem to elucidate the physiologicalfunctions of the Kyungrak systemrdquo Acetylcholine and pilo-carpine were used and the electrical potential changes wererecorded

The experiment result reported that the bioelectricalsignals changed after stimulation with acetylcholine pilo-carpine and Novocain He noted that the signals variedaccording to the different types and concentration of drugs(Figure 11) Finally the experiment of the bioelectrical signal

Figure 12 Strong electrical stimulation amplifies the changes in thebioelectrical activity of a Bonghan corpuscle

conduction of the Kyungrak system was based on his firstpaper which reported the interaction materially and func-tionally between Bonghan corpuscles and internal organs(Figure 12) [6] This study was significant in that it deter-mined how the stimulation signals of Bonghan corpuscleswere delivered in the Kyungrak system He stimulated Bong-han corpuscles connected to a Bonghan duct to measure thebioelectrical potential of the next Bonghan corpuscle Theexperimental result reported that the bioelectrical potentialchanged after a certain time and that the delivery speed ofthe effect stimuli was 30mmsec at a superficial Bonghancorpuscle (a Bonghan corpuscle in the skin)

The relationship betweenmechanical movements and thebioelectrical signals of Bonghan ducts was reported in histhird paper It started with an exploration of Bonghan ductsnot Bonghan corpuscles and developed this line of researchfrom previous results The properties of bioelectrical signalswere similar to those of a superficial Bonghan corpuscle(a Bonghan corpuscle in the skin) and the delivery speedwas 1ndash3mmsec for communication in both directions Heasserted that the phenomena of longitudinal periodic lateraland pulsatory mixed movement could be observed [25] Themovement delivery speed of the Bonghan duct was 01ndash06mmsec which was much slower than that of the bioelec-trical signals However the experimental results pertaining tothe relationship between the mechanical movement and bio-electrical changing were limited because the explanation ofthe experiment setup and data was insufficient Nonethelesshe insisted that there was convincing evidence of the flow ofliquid actively through Bonghan ducts Afterward his studyconcentrated on Sanal cells (primomicrocells) and the liquidthrough Bonghan ducts

4 Recent Bioelectrical and ElectrophysiologyResearch on the Primo Vascular System

Electrophysiological research on the primo vascular systemhalted after his third paper His fourth and fifth papers werefocused on flowing Bonghan liquids and Sanal cells throughBonghan corpuscles and ducts Park started to measure andanalyze the electrical potential of Bonghan corpuscles in the2000s (Figure 13) [37] Various experiments involving thestaining Bonghan corpuscles and ducts were attempted todistinguish them from other tissues [4 23 38ndash42] Lee et alreported that trypan blue was the most effective type of stain

Evidence-Based Complementary and Alternative Medicine 7Vo

ltage

(V)

Volta

ge (V

)

400 600 800 1000 1200 1400 1600 1800

Time (s)

Time (s)

032

034

Spontaneous activity

Resting potentialDs

Te

Ve

032

034

535 555 575

(a)

Volta

ge (V

)

Time (s)

0334

0336

2830 2840 2850 2860 2870 2880 2890 2900 2910

Ts

Vs

Ds

(b)

Volta

ge (V

)

Time (s)2148 2150 2152 2154 2156 2158 2160 2162 2164 2166 2168

0328

0327

0326

0325

0324

Spontaneous evoked spike

TsFs

Vs

(c)

Figure 13 Resting potential and spontaneous activity of the potential in a Bonghan corpuscle (BHC) (a) at themoment of themicrocapillaryinsertion into a cell membrane of a BHCThe potential decreases abruptly by about 38mV from the reference potential of the bath 119881

119889is the

potential drop The potential increased slowly to the resting potential by 67mV 119881119890is the small increase and 119879

119890is the time of the increase

(452 sec) The resting potential remained stable with fine background fluctuations and the irregular activity of spontaneously evoked spikesin the resting potential arose for a duration (119863

119904) of about 166 seconds (b) The spontaneous activity in the resting potential was examined

more closely The average amplitude (119881119904) is 11mV and the average period (119879

119904) is 08 sec (c) More magnified view of the spontaneous activity

A spike has an average half-width (119865119904) of 027 seconds

for these corpuscles and ducts in 2007 [4] The trypan bluestaining method was advanced by B C Lee Park attemptedto stain Bonghan corpuscles using the trypan blue stainingmethod and attempted to measure and analyze the electricalsignals from Bonghan corpuscles of the intestines of rats [37]Heworked on threemain research topicsmeasuring and ana-lyzing the resting potential and spontaneous action potentialusing an intracellular recording method He also observedthe changing electrical potential by stimulation with variousdrugs such as acetylcholine modeled the electrical signals byBVP (Bonhoeffer-Van der Pol) modeling and analyzed thefindings by fractal theory J H Choi C J Choi and Chodeveloped the electrophysiology of the primo vascular systemafter this research [43ndash45]

41 Measurement of the Resting Potential and SpontaneousBioelectrical Potential of Primo Nodes and Vessels Park rec-ognized that the measurement of electrical signals was veryimportant because if Bonghan corpuscles and ducts areconnected to each other and some liquid flows throughthe ducts a driving force is necessary to deliver the liquidAccording to this result a resting potential and spontaneousaction potential existed They reported experimental valuesof minus399 plusmn 155mV and 12 plusmn 06mV [37]

This result was highly significant as it was the first evi-dence that Bonghan corpuscles were composed of excitablecells and not types of fibrins and collagen fibers In thosedays almost every anatomical and histological expert insistedthat Bonghan corpuscles and ducts are just collagen fibers


00004

00002

0001 01 1 10

Am

plitu

de

Frequency (Hz)

Figure 14 Fast Fourier transform (FFT) of the membrane poten-tial In this particular experiment there were four values 062ndash084Hz 036Hz 005Hz and 002Hz appearing above the broadbackground

and types of fibrins So the corpuscles and ducts are notonly new structures but are nonfunctioning despite the factthat they may be new structures However his result showedthat the spontaneous action potential was observed thus thecorpuscle must have some special function

Moreover when the electrical signals were analyzed theprimary wave groups were 062ndash084Hz 036Hz 005Hzand 002Hz (Figure 14) It is interesting to note that this issimilar to the E (geu) F (neu) and G (deu) wave formsreported by Kim in the 1960s [6 24ndash27 34]

Choi developed a more precise experimental setup andexperiment thatmeasured the electrical potential fromBong-han ducts [44] He was an oriental medical doctor and aphysicist who thought that Bonghan corpuscles and ductswere not related to the Kyungrak system In addition hehypothesized that Bonghan ducts were types of fibrin andwere similar to lymphatic vessels by conducting a comparisonexperiment

However he observed the resting potential (119899 = 17) andspontaneous action potential (119899 = 2) of Bonghan ductsreporting a resting potential of minus100 plusmn 47mV from cellsembedded in the surfaces of Bonghan ducts He concludedthe bioelectrical signals from Bonghan ducts and lymphaticvessels were different and that it would be pointless to discussa comparison with fibrins This was an important resultwhich overturned his previous hypothesis representing thefirst report of the resting potential and spontaneous actionpotential of Bonghan ducts (Figure 15) [44 46] There wassome discussion about revising the nomenclature to expandthe study of the Bonghan system around that time (Table 2)

The intracellular recording method and the patch clampmethod were used to measure the bioelectrical signals fromBonghan corpuscles and ducts by Choi [44] He measuredthe electrical signals using a whole-cell slice-patch recordingmethod for primo nodes and an intracellular recordingmethod for primo vessels His result reported a resting

Table 2 Revised nomenclature for the Bonghan system

Before AfterBonghan system Primo vascular systemBonghan duct Primo vesselBonghan corpuscle Primo nodeBonghan liquor Primo fluidBonghan sanal Primo microcell

potential of ndash3660 plusmn 138mV of the primo nodes but nospontaneous action potential Small round cells are mostabundant in primo nodes On the basis of the current-voltage(I-V) relationships and kinetics of the outward currents thecells of primonodes could be grouped into four types Amongthese type I cells were the majority (69) [43] He reportedthat the resting potential of primo vessels was 210 plusmn 22mVand that there were two groups based on the resting potential(Type A (70) minus1313plusmn066mV and Type B (30) minus3864plusmn296mV) However he reported that there were no propertiesof the spontaneous action potential of primo nodes andvessels even if there were 2ndash4 cell groups (Figure 16) [43]

The Korea Institute of Oriental Medicine began researchon the primo vascular system in 2009 Lee et al reported amethod of distinguishing between tornmesentery and primovessels [47] Cho reported that there was the spontaneousaction potential of primo vessels on internal organs using anextracellular recording method The value of the potentialwas different from the action potential of a pacemaker ofintestinesTherewere two types of cell groups reported aswell[45]

Summarizing the research results pertaining to the rest-ing potential and spontaneous action potential of primonodes and vessels resting potential and spontaneous actionpotential of cells from primo nodes and vessels and severalcell type groups were discovered

This conclusion was similar to the report of B H Kimwho found that there were distinct bioelectrical signals fromthe tissues of Bonghan corpuscles and ducts also findingthat modern electrophysiological technology could increasethe confidence in results pertaining to the properties ofbioelectrical signals from the primo vascular system

42 Electrophysiological Study of the PrimoVascular System byResponding Drugs Park was interested in the changing bio-electrical signals of Bonghan corpuscles by drug stimulationafter measuring the resting potential and spontaneous actionpotential [37]

This was direct evidence that the Bonghan system maybe a circulatory system such as the cardiac vascular systemand the lymphatic system because if there were excitablecells of the Bonghan system that responded to drugs thesystem could contribute to the functions of the living bodylike nerves andmuscles He attempted to find the response ofbioelectrical signals from Bonghan corpuscles using acetyl-choline pilocarpine atropine and nifedipine Acetylcholineis a major neurotransmitter in an autonomic nervous systemIt stimulates both nicotinic and muscarinic receptors and


AcetylcholineWashing

Time (s)0 400 800 1200 1600

Time (s)760 800 840 880

Time (s)894 895 896

Mem

bran

e pot

entia

l (m

V)

Mem

bran

e pot

entia

l (m

V)

Mem

bran

e pot

entia

l (m

V)

minus25

minus20

minus15

minus10

minus5

minus25

minus20

minus15

minus10

minus5

minus40

minus20

0

Figure 15 Spontaneous action potentials of the novel thread part The resting potential was minus22mV this is a quite unique value comparedto that of other excitable cells The bursting frequency decreased after applying acetylcholine and increased slightly after washing

is a common drug used to stimulate muscles Pilocarpineis a nonselective muscarinic acetylcholine receptor agonistAtropine is a cholinergic receptor antagonist and a com-petitive nonselective antagonist at central and peripheralmuscarinic acetylcholine receptors Consequently as theelectrical signals of Bonghan corpuscles respond to thesedrugs Bonghan corpuscles have muscarinic receptors whichare controlled by the autonomic nerve system The Bonghansystem may be an autonomic-nerve-controlled system

Nifedipine is L-type Ca2+ channel blocker The funda-mental features of the spike generated in smooth muscles arerelated to the activation of the L-type Ca2+ channel Ca2+must be available for muscle contraction Therefore if thereare Ca2+ ion channels in Bonghan corpuscles it could beimportant evidence that Bonghan corpuscles have smoothmuscle-like characteristics such as contractibility and cellrelaxation

Park reported that the resting potential due to stimu-lation by acetylcholine was decreased to 50 and that thespike shape of the spontaneous action potentials completelychanged [37] Pilocarpine also changed the potential ofBonghan corpuscles similarly [48] Although the corpuscleswere stimulated by acetylcholine and pilocarpine the rest-ing potential of Bonghan corpuscles increased dramaticallywhen stimulated by atropine (Figure 17) [37] In the case ofnifedipine stimulation the resting potential also increased

though it decreased after stimulation with acetylcholine andpilocarpine This was the first evidence of the existenceof Ca2+ ion channels in the cell membranes of Bonghancorpuscles

This result confirms that the Bonghan system may havethe functions of contractibility and relaxation of the circula-tory system [37] Particularly Park asserted that the electricalsignals of smooth muscle-like cells from Bonghan corpuscleshave similar properties to those of the vascular smoothmuscle reported by Bkaily [49]

J H Choi studied the responses to drugs He usedtetraethylammonium (TEA) TEA is known to block the K+channel in nerves and Ca2+ channels are also blocked [50 51]

Tetraethylammonium (TEA) dose-dependently blockedboth the outward and inward current (IC

50 43mM at

plusmn60mV) Under current clamp conditions TEA dose-dependently depolarized the membrane potential (185mVat 30mM) with an increase in the input resistance Theseresults demonstrate for the first time that a TEA-sensitivecurrent with limited selectivity to K+ contributes to theresting membrane potential in type I cells [52]

In summary in the previous results of drug stimulationexperiments primo nodes are shown to have muscarinicreceptors allowing the primo vascular system to be con-trolled by the autonomic nerve system Moreover the systemhas the possibility of contractibility and relaxation function


Voltage (mV)

0

10

20

30

40

50

minus80 minus60 minus40 minus20 20 40 60 800minus10

Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(a)

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10

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50


Curr

ent d

ensit

y (p

Ap

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(b)

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ent d

ensit

y (p

Ap

F)

100pA100ms

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10

20

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50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(d)

Figure 16 Four types of current-voltage (IV) relationships recorded from cells in PN slices IV curves were obtained by depolarizing steppulses from minus80 to 80mV (a) Type IIV relationships showing outward rectification Note that the IV relationships measured at 50ms (solidsquares) and 550ms of the 600ms current pulse (open circle) are identical (b) Type IIIV relationships showing outward rectification withthe time-dependent activation of the outward current (c) Type III IV relationships showing outward rectification with a time-dependentand linearly activated outward current Note the lower current density in this cell (d) Type IV IV relationships showing outward rectificationwith a time-dependent and hyperbolic increase in the outward current The insets show the current traces for the IV relationships in (a) and(d) The holding potential is minus30 (a) and minus40mV (b and d) The largest outward tail current is seen in the current traces of the type IV cellScale bars for the insets are 10120583m

due to the Ca2+ ion channels Moreover the cells of the primovascular system may be excitable given the K+ ion channelsin the membranes of the cells These results are in goodagreementwith the proteomics analyses of the primo vascularsystem [53]

5 Discussion

The results of the bioelectrical signals and electrophysiologyresearch are summarized in Tables 3 and 4

The bioelectrical study of primo nodes and vessels sim-ply focused on the measurement of the resting potentialand spontaneous action potential initially as nobody knewwhether the cells from the primo vascular system had intrin-sic potentials or notTherefore it is important to confirm theproperties of the action potential of cells related to the basicphysiology functions

While examining the papers related to the bioelectricalstudy of the primo vascular system there were slight differ-ences in the results of the resting potential action potentialand the drugs responses depending on the studyThe researchis not sufficient for a complete understanding of the functionsof the primo vascular system in the body precisely but the

primo vascular system clearly has excitable cells that respondto some stimuli

The physiological functions of the primo vascular systemshould be confirmed by investigating the properties of eachcell in the system and by measuring the resting potential andthe action potential of each cell type If each cell type of theprimo vascular system could be determined wewill be able tounderstand the signal delivery methods and the conductionsystem

Ultimately the purpose of an electrical signal analysisof the primo vascular system is to investigate whether theinternal substances of the primo nodes and primo vesselscan be circulated In addition it is necessary to conductadditional experiments to answer the questions pertainingto the existence of an exclusive signal conduction system ofthe primo system circulation like the nervous system andto find evidence supporting the circulating function of theprimo system based on actual mechanical activity Researchareas requested in the future may include the following

(1) standardization of measured data by the standardiza-tion of techniques to measure the electrical signals ofthe primo vascular system


037

036

035

034

033

032

031

5700 5800 5900 6000 6100 6200Time (s)

Volta

ge (V

)

Atropine (1120583M)

V998400R

D998400s

TR

ΔVRVRPilocarpine (15120583M)

(a)

6150 6200 6250Time (s)

0396

0352

0348

Volta

ge (V

)

D998400s

(b)

6150 6200 6250Time (s)

03535

0354

03545

0355

03555

Volta

ge (V

)

F998400s T998400

s

V998400s

(c)

Figure 17 Effects of atropine on the resting potential and the spontaneous burst (a) The resting potential dropped from 119881119877to 1198811015840119877and the

decrease was Δ119881119877 The resting potential slowly recovered its original value by Δ119881

119877 (b) The duration of the spontaneous burst (1198631015840

119904) (c) The

amplitude (1198811015840119904) period (1198791015840

119904) and half width (1198651015840

119904) of the spikes in the burst dramatically changed

(2) characterizing the cells composing the primo vascularsystem

(3) investigating the electrical signal conduction andmechanical activity of the primo vascular system

(4) uncovering the signal conduction mechanismthrough a connection between the primo vascularsystem and internal organs materially

(5) investigating the control of the primo vascular systemby external stimulation mechanisms such as medica-tions and their interrelationships

(6) looking into the circulation of internal substancesinside the primo vascular system

In addition it is recommended to adopt useful technology tomeasure two-dimension spatial signal conduction methodssuch as optical electrophysiological techniques which have

greatly progressed recently in addition to previous tech-niques of measuring one-dimension signals from a singlepoint [54]

Though research on the electrical signals of the primo sys-tem started in the 1960s the reliability of the study results waslimited due to the limitations of the measuring technologyand short descriptions of the methods and data Howeverseveral studies of the electrophysiological characteristics ofthe primo vascular system have been performed since the2000s and it will be interesting to watch the development inthe field of the electrophysiology of the primo vascular systemin the future

In conclusion there are many differenct type cells com-posed of primo vascular system one is the excitable cell typeand the other is the nonexcitable cell type

In case of the excitable cell the vascular smooth musclelike cell type [37] the intestinal smooth muscle like cell type[45] and the immune cell type are reported [46]


Table 3 A comparison of the electrophysiological characteristics between primo nodes primo vessels neurons and muscle cells

RP (mV) APD (mV) Drug-responsive ReferenceNeuron minus70 1 Y [19]Muscle

Skeletal minus80 to minus90 2 to 5 Y [20]Cardiac minus85 to minus95 200 to 400 Y [21]Smooth minus50 to minus60 10 to 50 Y [22]

Primo nodeSkin ∙ 01 Y [25]Surface of a small intestine minus399 to 155 12 to 06 Y [37]Surface of a small intestine minus366 to 138 ∙ Y [43]

Primo vesselSkin ∙ 01 Y [25]

Surface of a small intestine minus10 to 47 2040 ∙ [44]

Surface of a small intestine A minus131 to 07 Bminus386 to 30 ∙ Y [43]

Surface of a small intestine ∙A 100 to 84B 137 to 87 ∙ [45]

RP resting potential TP threshold potential APD action potential duration (bioelectrical activity)

Table 4 The summarization of the electrophysiological properties of the primo vascular system

Target Methodology Cell Type Result Reference

Primo node

Intracellular recording Vascular smooth musclelike

Excitable(Muscarinic receptor Ca2+ ion channel) [37]

Patch clampCurrent-voltage relationship Four types Nonexcitable cell

(K+ ion channel for Type I) [43]

Extracelluar recording Two types Excitable cell(not smooth muscle like) [45]

Primo duct Intracellular recordingSmooth muscleSecreting cellImmune cell

ExcitableNon-excitable [44]

Acknowledgments

This work was supported by a Grant from the Ministry ofKnowledge Economy (no 10042250) and byKIOM(K13290)

References

[1] H-S Shin H-M Johng B-C Lee et al ldquoFeulgen reactionstudy of novel threadlike structures (Bonghan ducts) on thesurfaces of mammalian organsrdquo Anatomical Record B vol 284no 1 pp 35ndash40 2005

[2] B-C Lee and K-S Soh ldquoContrast-enhancing optical methodto observe a Bonghan duct floating inside a lymph vessel of arabbitrdquo Lymphology vol 41 no 4 pp 178ndash185 2008

[3] B-C Lee H B Kim B Sung et al ldquoNetwork of endocardialvesselsrdquo Cardiology vol 118 no 1 pp 1ndash7 2011

[4] B-C Lee S Kim and K-S Soh ldquoNovel anatomic structuresin the brain and spinal cord of rabbit that may belong to theBonghan systemof potential acupuncturemeridiansrdquo Journal ofAcupuncture andMeridian Studies vol 1 no 1 pp 29ndash35 2008

[5] K-S Soh ldquoBonghan duct and acupuncture meridian as opticalchannel of biophotonrdquo Journal of the Korean Physical Societyvol 45 no 5 pp 1196ndash1198 2004

[6] B H Kim ldquoStudy on the reality of acupuncture meridiansrdquoJournal of Jo Sun Medicine vol 9 pp 5ndash13 1962

[7] K-S Soh ldquoBonghan circulatory system as an extension ofacupuncture meridiansrdquo Journal of Acupuncture and MeridianStudies vol 2 no 2 pp 93ndash106 2009

[8] LGalvaniDeViribus Electricitatis inMotuMusculari Commen-tarius Ex Typographia Instituti Scientiarium Bologna Italy1791

[9] D AWellsThe Science of CommonThings a Familiar Explana-tion of the First Principles of Physical Science Ivision BlakemanTaylor New York NY USA 1872

[10] J Malmivuo and R Plonsey Bioelectromagnetism Principlesand Applications of Bioelectric and Biomagnetic Fields OxfordUniversity Press New York NY USA 1995

[11] A L Hodgkin and A F Huxley ldquoA quantitative descriptionof membrane current and its application to conduction and


excitation in nerverdquoThe Journal of Physiology vol 117 no 4 pp500ndash544 1952

[12] J Graham and R W Gerard ldquoMembrane potentials andexcitation of impaled single muscle fibersrdquo Journal of CellularPhysiology vol 28 no 1 pp 99ndash117 1946

[13] E Neher and B Sakmann ldquoSingle channel currents recordedfrom membrane of denervated frog muscle fibersrdquo Nature vol260 no 5554 pp 799ndash802 1976

[14] A Huxley ldquoFrom overshoot to voltage clamprdquo Trends inNeurosciences vol 25 no 11 pp 553ndash558 2002

[15] M Scanziani and M Hausser ldquoElectrophysiology in the age oflightrdquo Nature vol 461 no 7266 pp 930ndash939 2009

[16] E Neher B Sakmann and J H Steinbach ldquoThe extracellularpatch clamp a method for resolving currents through individ-ual open channels in biological membranesrdquo Pflugers Archivvol 375 no 2 pp 219ndash228 1978

[17] WWalz Patch-Clamp Analysis Advanced Techniques HumanaPress Totowa NJ USA 2nd edition 2007

[18] J T Fulton Biological Vision a 21st Century Tutorial TraffordVictoria Canada 2004

[19] N R Carlson Foundations of Physiological Psychology Allynand Bacon Boston Mass USA 2nd edition 1992

[20] F Bezanilla ldquoThe action potential from voltage-gated conduc-tances to molecular structuresrdquo Biological Research vol 39 no3 pp 425ndash435 2006

[21] R E Klabunde Cardiovascular Physiology Concepts LippincottWilliams ampWilkins Philadelphia Pa USA 2005

[22] A Corrias and M L Buist ldquoA quantitative model of gastricsmooth muscle cellular activationrdquo Annals of Biomedical Engi-neering vol 35 no 9 pp 1595ndash1607 2007

[23] B-C Lee S-U Jhang J-H Choi S-Y Lee P-D Ryu and K-SSoh ldquoDiI staining of fine branches of Bonghan ducts on surfaceof rat abdominal organsrdquo Journal of Acupuncture and MeridianStudies vol 2 no 4 pp 301ndash305 2009

[24] B H Kim ldquoOn the acupuncture meridian systemrdquo Journal of JoSun Medicine vol 90 pp 6ndash35 1963

[25] B H Kim ldquoOn the Kyungrak systemrdquo Journal of the Academyof Medical Sciences of the Democratic Peoplersquos Republic of Koreavol 90 pp 1ndash41 1963

[26] B H Kim ldquoSanal and hematopoiesisrdquo Journal of Jo SunMedicine vol 108 pp 1ndash6 1965

[27] B H Kim ldquoSanal and theoryrdquo Journal of Jo Sun Medicine vol108 pp 39ndash62 1965

[28] N Jeh ldquoLe dereglement de lrsquoequilibre de lrsquoenergie dans unmeridien considere comme lrsquoetiologie de certains types drsquoalgiesrdquoBulletin de la Societe drsquoAcupuncture vol 20 pp 58ndash70 1956

[29] C-R Overhof Uber das Elektrische Verhalten Spezieller Haut-bezirke 1960

[30] H Ogata T Matsumoto and H Tsukahara ldquoElectrical skinresistance changes in meridians during ophthalmic surgerywith local anesthesiardquo American Journal of Chinese Medicinevol 11 no 1ndash4 pp 130ndash136 1983

[31] R Voll Elektroakupunktur Anderthalb Jahrzehnte Forschungund Erfahrung in Diagnostik undTherapie 1971

[32] A C Ahn A P Colbert B J Anderson et al ldquoElectricalproperties of acupuncture points and meridians a systematicreviewrdquo Bioelectromagnetics vol 29 no 4 pp 245ndash256 2008

[33] S D Mist M Aickin P Kalnins et al ldquoReliability of AcuGraphsystem formeasuring skin conductance at acupointsrdquoAcupunc-ture in Medicine vol 29 no 3 pp 221ndash226 2011

[34] B H Kim ldquoThe Kyungrak systemrdquo Journal of Jo Sun Medicinevol 108 pp 1ndash38 1965

[35] ldquoDonguibogam Principles and Practice of Eastern MedicinerdquoUNESCO registered

[36] B Sakmann and E Neher ldquoPatch clamp techniques for studyingionic channels in excitable membranesrdquo Annual Review ofPhysiology vol 46 pp 455ndash472 1984

[37] S H Park Bioelectrical study of Bonghan system [PhD thesis]Seoul National University 2009

[38] B-C Lee K Y Baik H-M Johng et al ldquoAcridine orangestaining method to reveal the characteristic features of anintravascular threadlike structurerdquo Anatomical Record B vol278 no 1 pp 27ndash30 2004

[39] B-C Lee J S Yoo K Y Baik K W Kim and K-S Soh ldquoNovelthreadlike structures (Bonghan ducts) inside lymphatic vesselsof rabbits visualized with a Janus Green B staining methodrdquoAnatomical Record B vol 286 no 1 pp 1ndash7 2005

[40] C Lee S-K Seol B-C Lee Y-K Hong J-H Je and K-SSoh ldquoAlcian blue stainingmethod to visualize Bonghan threadsinside large caliber lymphatic vessels and X-ray microtomog-raphy to reveal their microchannelsrdquo Lymphatic Research andBiology vol 4 no 4 pp 181ndash190 2006

[41] B-C Lee and K-S Soh ldquoVisualization of acupuncture merid-ians in the hypodermis of rat using Trypan bluerdquo Journal ofAcupuncture andMeridian Studies vol 3 no 1 pp 49ndash52 2010

[42] H S Lee W H Park A R Je H S Kweon and B C LeeldquoEvidence for novel structures (primo vessels and primo nodes)floating in the venous sinuses of rat brainsrdquoNeuroscience Lettersvol 522 no 2 pp 98ndash102 2012

[43] J H Choi Electrophysiological properties of cells in the organsurface primo-vascular system of rats [PhD thesis] SeoulNational University 2011

[44] C J Choi Spontaneous action potential and basic eletricalcharacteristics of cells in the Bonghan duct [PhD thesis] SeoulNational University 2009

[45] S J Cho S H Lee W Zhang et al ldquoMathematical distinctionin action potential between primo-vessels and smooth musclerdquoEvidence-Based Complementary and Alternative Medicine vol2012 Article ID 269397 6 pages 2012

[46] C-J Choi J H Jung K-S Soh Y H Ryu P D Ryu and S-HPark ldquoSpontaneous action potential from threadlike structureson the surfaces of abdominal organsrdquo Journal of the KoreanPhysical Society vol 58 no 4 pp 831ndash836 2011

[47] S Lee Y Ryu Y Yun et al ldquoAnatomical discrimination of thedifferences between torn mesentery tissue and internal organ-surface primo-vesselsrdquo Journal of Acupuncture and MeridianStudies vol 3 no 1 pp 10ndash15 2010

[48] S H L B C Park C J Choi K S Soh and P D Ryu ldquoEffectsof cholinergic drugs on membrane potential of cells in organsurface primo nodesrdquo in The Primo Vascular System pp 251ndash261 2012

[49] G Bkaily Ionic Channels in Vascular Smooth Muscle RGLandes Austin Tex USA 1994

[50] R Latorre C Vergara and C Hidalgo ldquoReconstitution inplanar lipid bilayers of a Ca2+-dependent K+ channel fromtransverse tubule membranes isolated from rabbit skeletalmusclerdquo Proceedings of the National Academy of Sciences of theUnited States of America vol 79 no 3 pp 805ndash809 1982

[51] D G Lang and A K Ritchie ldquoTetraethylammonium blockadeof apamin-sensitive and insensitive Ca2+-activated K+ chan-nels in a pituitary cell linerdquo The Journal of Physiology vol 425pp 117ndash132 1990


[52] J-H Choi C J Lim T H Han S K Lee S Y Lee andP D Ryu ldquoTEA-sensitive currents contribute to membranepotential of organ surface primo-node cells in ratsrdquoThe Journalof Membrane Biology vol 239 no 3 pp 167ndash175 2011

[53] S J Lee B C Lee C H Nam et al ldquoProteomic analysisfor tissues and liquid from Bonghan ducts on rabbit intestinalsurfacesrdquo Journal of Acupuncture and Meridian Studies vol 1no 2 pp 97ndash109 2008

[54] T J Ebner and G Chen ldquoUse of voltage-sensitive dyes andoptical recordings in the central nervous systemrdquo Progress inNeurobiology vol 46 no 5 pp 463ndash506 1995


C CopperZ ZincN Nerve

C

N

Z

Figure 1 The electrical stimulation of a frog nerve It was foundthat the current flows through the two different metals coming intocontact with animal muscle

evidence showing that the primo vascular system may be aDNA circulatory system has been reported as well [4]

Electrophysiology is a science of the electrical propertiesof biological tissues and cells It involves measurements ofvoltage changes or electric current on a wide variety of scalesfrom single-ion channel proteins to whole organs such as theheart In this paper we introduce the history of electricalsignals and the electrophysiology of the primo vascularsystem

Section 2 introduces the understanding of electrophysi-ology and modern techniques to measure the bioelectricalsignal for general readers Section 3 describes the beginningof the bioelectrical study of acupuncturemeridian briefly andreviews the electrophysiological study of the primo vascularsystem past and present The last section arranges the resultof the reviewed papers and suggests the future work for theelectrophysiology of the primo vascular system

2 Development of Electrophysiology withElectrical Signal and Basic Medicine

21 Bioelectrical Signal Studies and Electrophysiology In 1791the Italian physician andphysicist LuigiGalvani first recordedthe phenomenon of electrical signals while dissecting a frogon a table where he had been conducting experiments withstatic electricity (Figure 1) Galvani coined the term animalelectricity to describe the phenomenon while contempo-raries labeled it galvanism Galvani and his contemporariesregarded muscle activation as resulting from an electricalfluid or substance in the nerves [8 9]

In the work to study cardiac electrophysiology in orderto gain a better understanding of bioelectricity cardiacelectrophysiology emerged as an important area to elucidateand diagnose the heart function and to treat heart disease

including arrhythmia through electrocardiograms (Figure 2)[10] The function of tissues and organs in humans is closelyrelated to analyses of the bioelectrical signals from thesefeatures

22 Development of a Measurement Method for BioelectricSignals Electrophysiological studies which started fromfrog muscle response research extended the fundamentalknowledge of nerve cells through squid axon membranepotential measurements using an electrode when Hodgkinand Huxley began this work in 1952 [11] Graham and Gerardsucceeded to create and develop an electrode with a finerdiameter of 2ndash5 um to measure the intracellular potential(Figure 3) [12] With the development of this measurementtechnique Neher and Sakmann reduced the signal-to-noiseratio using a seamless seal between the cell membrane andthe electrode establishing the foundation of the study of ionchannels in the cell membrane [13] Cell membrane potentialmeasurement techniques in electrophysiological research areoutlined later

221 Intracellular Recording (Voltage Clamp and CurrentClamp) An electrical signal representing cell activities isa result of the activity of the ion channels in the cellmembrane The cell activity is associated with whether thecell membrane ion channel is activating or not and is thenrelated to the electrical signals from the cells Related to thisthe voltage clamp technique was developed to measure theintracellular ion flow with the constant membrane potentialThis technique was useful in research on the mechanism ofbiological electrical signals generated by the ions such as K+or Na+ passing through the electrically opened and closedchannels [11ndash13]

In contrast the current clamp was used to measure thechanges of the voltage inside the cell by means of a constantcurrent this was useful to classify cell types by understandingthe action potential of the cells (Figure 4) [14 15]

222 Patch Clamp As an intracellular recording techniquewhich involves inserting an electrode into a cell directlythe patch clamp technique was created for the purpose ofseparating and analyzing of each ion channel existing onthe membrane of a cell Patch clamp recordings use as anelectrode a glass micropipette that has an open tip diameterof about one micrometer (1120583m) which is a size enclosinga membrane surface area or ldquopatchrdquo that often contains justone or few ion channel molecules This type of electrode isdistinct from the ldquosharp microelectroderdquo used to impale cellsin traditional intracellular recordings in that it is sealed ontothe surface of the cell membrane rather than inserted throughit Many researchers including Neher and Sakmann studiedion channels in various cells using the patch clamp technique(Figure 5) [16ndash18]

They observed that the electrophysiological characteris-tics in each major tissue cell especially the resting potentialshowed a different form (Table 1) This cell-size level tech-nique led to the development of electrophysiology and has


0 100 200 300 400 500 600 700Time (ms)

Sinusnode

Atrialmuscle

A-Vnode

CommonbundleBundle


Ventricularmuscle

P

Q

R

S

TU


Cell

Voltageelectrode











Action potential

Depolarization

Hyperpolarization

1 2 3Time (ms)

Mem

bran

e pot

entia

l (m

V)


Thre

shol

d ex

cita

tion

5 4 3 2 1

Stimulusapplied

minus90

minus80

minus70

minus60

minus50

minus40

minus30

minus20

minus10

010

20

30

40

(a)

Time (ms)0 2 4 6 8 10

10

20

30

40

50

60

70

0

20

40

60Action potential


minus60

minus40

minus20V(m

V)

G(m

scm

2)

Na+ conduction

K+ conduction

(b)


50

0

minus50

minus100

(mV

)

4

0

12

3

4

(c)

Mem

bran

e pot

entia

l (m

V)

Time (ms)0 2 4 6


(times104)

minus70

minus60

minus50

minus40

(d)









On-cellpatch

Inside-outpatch

Whole-cellclamp

Outside-outpatch

(a) (b)

(c) (d)















potentialSpike























ltage

(V)

Volta

ge (V

)

400 600 800 1000 1200 1400 1600 1800

Time (s)

Time (s)

032

034


Resting potentialDs

Te

Ve

032

034

535 555 575

(a)

Volta

ge (V

)

Time (s)

0334

0336

2830 2840 2850 2860 2870 2880 2890 2900 2910

Ts

Vs

Ds

(b)

Volta

ge (V

)

Time (s)2148 2150 2152 2154 2156 2158 2160 2162 2164 2166 2168

0328

0327

0326

0325

0324


TsFs

Vs

(c)


119889is the












00004

00002

0001 01 1 10

Am

plitu

de

Frequency (Hz)

















Time (s)0 400 800 1200 1600

Time (s)760 800 840 880

Time (s)894 895 896

Mem

bran

e pot

entia

l (m

V)

Mem

bran

e pot

entia

l (m

V)

Mem

bran

e pot

entia

l (m

V)

minus25

minus20

minus15

minus10

minus5

minus25

minus20

minus15

minus10

minus5

minus40

minus20

0









50 43mM at




Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(a)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(b)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

100pA100ms

(c)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(d)



5 Discussion









037

036

035

034

033

032

031

5700 5800 5900 6000 6100 6200Time (s)

Volta

ge (V

)

Atropine (1120583M)

V998400R

D998400s

TR


(a)

6150 6200 6250Time (s)

0396

0352

0348

Volta

ge (V

)

D998400s

(b)

6150 6200 6250Time (s)

03535

0354

03545

0355

03555

Volta

ge (V

)

F998400s T998400

s

V998400s

(c)




119904) (c) The


119904) and half width (1198651015840
























Primo node








Acknowledgments


References



























































0 100 200 300 400 500 600 700Time (ms)

Sinusnode

Atrialmuscle

A-Vnode

CommonbundleBundle


Ventricularmuscle

P

Q

R

S

TU


Cell

Voltageelectrode











Action potential

Depolarization

Hyperpolarization

1 2 3Time (ms)

Mem

bran

e pot

entia

l (m

V)


Thre

shol

d ex

cita

tion

5 4 3 2 1

Stimulusapplied

minus90

minus80

minus70

minus60

minus50

minus40

minus30

minus20

minus10

010

20

30

40

(a)

Time (ms)0 2 4 6 8 10

10

20

30

40

50

60

70

0

20

40

60Action potential


minus60

minus40

minus20V(m

V)

G(m

scm

2)

Na+ conduction

K+ conduction

(b)


50

0

minus50

minus100

(mV

)

4

0

12

3

4

(c)

Mem

bran

e pot

entia

l (m

V)

Time (ms)0 2 4 6


(times104)

minus70

minus60

minus50

minus40

(d)









On-cellpatch

Inside-outpatch

Whole-cellclamp

Outside-outpatch

(a) (b)

(c) (d)















potentialSpike























ltage

(V)

Volta

ge (V

)

400 600 800 1000 1200 1400 1600 1800

Time (s)

Time (s)

032

034


Resting potentialDs

Te

Ve

032

034

535 555 575

(a)

Volta

ge (V

)

Time (s)

0334

0336

2830 2840 2850 2860 2870 2880 2890 2900 2910

Ts

Vs

Ds

(b)

Volta

ge (V

)

Time (s)2148 2150 2152 2154 2156 2158 2160 2162 2164 2166 2168

0328

0327

0326

0325

0324


TsFs

Vs

(c)


119889is the












00004

00002

0001 01 1 10

Am

plitu

de

Frequency (Hz)

















Time (s)0 400 800 1200 1600

Time (s)760 800 840 880

Time (s)894 895 896

Mem

bran

e pot

entia

l (m

V)

Mem

bran

e pot

entia

l (m

V)

Mem

bran

e pot

entia

l (m

V)

minus25

minus20

minus15

minus10

minus5

minus25

minus20

minus15

minus10

minus5

minus40

minus20

0









50 43mM at




Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(a)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(b)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

100pA100ms

(c)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(d)



5 Discussion









037

036

035

034

033

032

031

5700 5800 5900 6000 6100 6200Time (s)

Volta

ge (V

)

Atropine (1120583M)

V998400R

D998400s

TR


(a)

6150 6200 6250Time (s)

0396

0352

0348

Volta

ge (V

)

D998400s

(b)

6150 6200 6250Time (s)

03535

0354

03545

0355

03555

Volta

ge (V

)

F998400s T998400

s

V998400s

(c)




119904) (c) The


119904) and half width (1198651015840
























Primo node








Acknowledgments


References



























































Action potential

Depolarization

Hyperpolarization

1 2 3Time (ms)

Mem

bran

e pot

entia

l (m

V)


Thre

shol

d ex

cita

tion

5 4 3 2 1

Stimulusapplied

minus90

minus80

minus70

minus60

minus50

minus40

minus30

minus20

minus10

010

20

30

40

(a)

Time (ms)0 2 4 6 8 10

10

20

30

40

50

60

70

0

20

40

60Action potential


minus60

minus40

minus20V(m

V)

G(m

scm

2)

Na+ conduction

K+ conduction

(b)


50

0

minus50

minus100

(mV

)

4

0

12

3

4

(c)

Mem

bran

e pot

entia

l (m

V)

Time (ms)0 2 4 6


(times104)

minus70

minus60

minus50

minus40

(d)









On-cellpatch

Inside-outpatch

Whole-cellclamp

Outside-outpatch

(a) (b)

(c) (d)















potentialSpike























ltage

(V)

Volta

ge (V

)

400 600 800 1000 1200 1400 1600 1800

Time (s)

Time (s)

032

034


Resting potentialDs

Te

Ve

032

034

535 555 575

(a)

Volta

ge (V

)

Time (s)

0334

0336

2830 2840 2850 2860 2870 2880 2890 2900 2910

Ts

Vs

Ds

(b)

Volta

ge (V

)

Time (s)2148 2150 2152 2154 2156 2158 2160 2162 2164 2166 2168

0328

0327

0326

0325

0324


TsFs

Vs

(c)


119889is the












00004

00002

0001 01 1 10

Am

plitu

de

Frequency (Hz)

















Time (s)0 400 800 1200 1600

Time (s)760 800 840 880

Time (s)894 895 896

Mem

bran

e pot

entia

l (m

V)

Mem

bran

e pot

entia

l (m

V)

Mem

bran

e pot

entia

l (m

V)

minus25

minus20

minus15

minus10

minus5

minus25

minus20

minus15

minus10

minus5

minus40

minus20

0









50 43mM at




Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(a)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(b)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

100pA100ms

(c)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(d)



5 Discussion









037

036

035

034

033

032

031

5700 5800 5900 6000 6100 6200Time (s)

Volta

ge (V

)

Atropine (1120583M)

V998400R

D998400s

TR


(a)

6150 6200 6250Time (s)

0396

0352

0348

Volta

ge (V

)

D998400s

(b)

6150 6200 6250Time (s)

03535

0354

03545

0355

03555

Volta

ge (V

)

F998400s T998400

s

V998400s

(c)




119904) (c) The


119904) and half width (1198651015840
























Primo node








Acknowledgments


References



























































On-cellpatch

Inside-outpatch

Whole-cellclamp

Outside-outpatch

(a) (b)

(c) (d)















potentialSpike























ltage

(V)

Volta

ge (V

)

400 600 800 1000 1200 1400 1600 1800

Time (s)

Time (s)

032

034


Resting potentialDs

Te

Ve

032

034

535 555 575

(a)

Volta

ge (V

)

Time (s)

0334

0336

2830 2840 2850 2860 2870 2880 2890 2900 2910

Ts

Vs

Ds

(b)

Volta

ge (V

)

Time (s)2148 2150 2152 2154 2156 2158 2160 2162 2164 2166 2168

0328

0327

0326

0325

0324


TsFs

Vs

(c)


119889is the












00004

00002

0001 01 1 10

Am

plitu

de

Frequency (Hz)

















Time (s)0 400 800 1200 1600

Time (s)760 800 840 880

Time (s)894 895 896

Mem

bran

e pot

entia

l (m

V)

Mem

bran

e pot

entia

l (m

V)

Mem

bran

e pot

entia

l (m

V)

minus25

minus20

minus15

minus10

minus5

minus25

minus20

minus15

minus10

minus5

minus40

minus20

0









50 43mM at




Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(a)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(b)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

100pA100ms

(c)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(d)



5 Discussion









037

036

035

034

033

032

031

5700 5800 5900 6000 6100 6200Time (s)

Volta

ge (V

)

Atropine (1120583M)

V998400R

D998400s

TR


(a)

6150 6200 6250Time (s)

0396

0352

0348

Volta

ge (V

)

D998400s

(b)

6150 6200 6250Time (s)

03535

0354

03545

0355

03555

Volta

ge (V

)

F998400s T998400

s

V998400s

(c)




119904) (c) The


119904) and half width (1198651015840
























Primo node








Acknowledgments


References









































































ltage

(V)

Volta

ge (V

)

400 600 800 1000 1200 1400 1600 1800

Time (s)

Time (s)

032

034


Resting potentialDs

Te

Ve

032

034

535 555 575

(a)

Volta

ge (V

)

Time (s)

0334

0336

2830 2840 2850 2860 2870 2880 2890 2900 2910

Ts

Vs

Ds

(b)

Volta

ge (V

)

Time (s)2148 2150 2152 2154 2156 2158 2160 2162 2164 2166 2168

0328

0327

0326

0325

0324


TsFs

Vs

(c)


119889is the












00004

00002

0001 01 1 10

Am

plitu

de

Frequency (Hz)

















Time (s)0 400 800 1200 1600

Time (s)760 800 840 880

Time (s)894 895 896

Mem

bran

e pot

entia

l (m

V)

Mem

bran

e pot

entia

l (m

V)

Mem

bran

e pot

entia

l (m

V)

minus25

minus20

minus15

minus10

minus5

minus25

minus20

minus15

minus10

minus5

minus40

minus20

0









50 43mM at




Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(a)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(b)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

100pA100ms

(c)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(d)



5 Discussion









037

036

035

034

033

032

031

5700 5800 5900 6000 6100 6200Time (s)

Volta

ge (V

)

Atropine (1120583M)

V998400R

D998400s

TR


(a)

6150 6200 6250Time (s)

0396

0352

0348

Volta

ge (V

)

D998400s

(b)

6150 6200 6250Time (s)

03535

0354

03545

0355

03555

Volta

ge (V

)

F998400s T998400

s

V998400s

(c)




119904) (c) The


119904) and half width (1198651015840
























Primo node








Acknowledgments


References



























































ltage

(V)

Volta

ge (V

)

400 600 800 1000 1200 1400 1600 1800

Time (s)

Time (s)

032

034


Resting potentialDs

Te

Ve

032

034

535 555 575

(a)

Volta

ge (V

)

Time (s)

0334

0336

2830 2840 2850 2860 2870 2880 2890 2900 2910

Ts

Vs

Ds

(b)

Volta

ge (V

)

Time (s)2148 2150 2152 2154 2156 2158 2160 2162 2164 2166 2168

0328

0327

0326

0325

0324


TsFs

Vs

(c)


119889is the












00004

00002

0001 01 1 10

Am

plitu

de

Frequency (Hz)

















Time (s)0 400 800 1200 1600

Time (s)760 800 840 880

Time (s)894 895 896

Mem

bran

e pot

entia

l (m

V)

Mem

bran

e pot

entia

l (m

V)

Mem

bran

e pot

entia

l (m

V)

minus25

minus20

minus15

minus10

minus5

minus25

minus20

minus15

minus10

minus5

minus40

minus20

0









50 43mM at




Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(a)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(b)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

100pA100ms

(c)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(d)



5 Discussion









037

036

035

034

033

032

031

5700 5800 5900 6000 6100 6200Time (s)

Volta

ge (V

)

Atropine (1120583M)

V998400R

D998400s

TR


(a)

6150 6200 6250Time (s)

0396

0352

0348

Volta

ge (V

)

D998400s

(b)

6150 6200 6250Time (s)

03535

0354

03545

0355

03555

Volta

ge (V

)

F998400s T998400

s

V998400s

(c)




119904) (c) The


119904) and half width (1198651015840
























Primo node








Acknowledgments


References



























































00004

00002

0001 01 1 10

Am

plitu

de

Frequency (Hz)

















Time (s)0 400 800 1200 1600

Time (s)760 800 840 880

Time (s)894 895 896

Mem

bran

e pot

entia

l (m

V)

Mem

bran

e pot

entia

l (m

V)

Mem

bran

e pot

entia

l (m

V)

minus25

minus20

minus15

minus10

minus5

minus25

minus20

minus15

minus10

minus5

minus40

minus20

0









50 43mM at




Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(a)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(b)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

100pA100ms

(c)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(d)



5 Discussion









037

036

035

034

033

032

031

5700 5800 5900 6000 6100 6200Time (s)

Volta

ge (V

)

Atropine (1120583M)

V998400R

D998400s

TR


(a)

6150 6200 6250Time (s)

0396

0352

0348

Volta

ge (V

)

D998400s

(b)

6150 6200 6250Time (s)

03535

0354

03545

0355

03555

Volta

ge (V

)

F998400s T998400

s

V998400s

(c)




119904) (c) The


119904) and half width (1198651015840
























Primo node








Acknowledgments


References




























































Time (s)0 400 800 1200 1600

Time (s)760 800 840 880

Time (s)894 895 896

Mem

bran

e pot

entia

l (m

V)

Mem

bran

e pot

entia

l (m

V)

Mem

bran

e pot

entia

l (m

V)

minus25

minus20

minus15

minus10

minus5

minus25

minus20

minus15

minus10

minus5

minus40

minus20

0









50 43mM at




Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(a)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(b)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

100pA100ms

(c)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(d)



5 Discussion









037

036

035

034

033

032

031

5700 5800 5900 6000 6100 6200Time (s)

Volta

ge (V

)

Atropine (1120583M)

V998400R

D998400s

TR


(a)

6150 6200 6250Time (s)

0396

0352

0348

Volta

ge (V

)

D998400s

(b)

6150 6200 6250Time (s)

03535

0354

03545

0355

03555

Volta

ge (V

)

F998400s T998400

s

V998400s

(c)




119904) (c) The


119904) and half width (1198651015840
























Primo node








Acknowledgments


References



























































Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(a)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(b)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

100pA100ms

(c)

Voltage (mV)

0

10

20

30

40

50


Curr

ent d

ensit

y (p

Ap

F)

200pA100ms

(d)



5 Discussion









037

036

035

034

033

032

031

5700 5800 5900 6000 6100 6200Time (s)

Volta

ge (V

)

Atropine (1120583M)

V998400R

D998400s

TR


(a)

6150 6200 6250Time (s)

0396

0352

0348

Volta

ge (V

)

D998400s

(b)

6150 6200 6250Time (s)

03535

0354

03545

0355

03555

Volta

ge (V

)

F998400s T998400

s

V998400s

(c)




119904) (c) The


119904) and half width (1198651015840
























Primo node








Acknowledgments


References



























































037

036

035

034

033

032

031

5700 5800 5900 6000 6100 6200Time (s)

Volta

ge (V

)

Atropine (1120583M)

V998400R

D998400s

TR


(a)

6150 6200 6250Time (s)

0396

0352

0348

Volta

ge (V

)

D998400s

(b)

6150 6200 6250Time (s)

03535

0354

03545

0355

03555

Volta

ge (V

)

F998400s T998400

s

V998400s

(c)




119904) (c) The


119904) and half width (1198651015840
























Primo node








Acknowledgments


References






































































Primo node








Acknowledgments


References












































































































Documents

History of Bioelectrical Study and the Electrophysiology ...€¦ · Evidence-BasedComplementaryandAlternativeMedicine 7 Voltage (V) Voltage (V) 400 600 800 1000 1200 1400 1600 1800