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Ana Margarida Araújo
Ana Rita Oliveira
Felicia Negru
Filipa Leal
Luís Rita
Professora Ana Sebastião 1
Nervous Impulse Transmission
3
4
5
Synapse
Electric
Chemical
7
Chemical Synapses
Action Potential is transmitted by
special proteins called
neurotransmitters.
8
Neurotransmitters Releasing
Neurotransmitters get out from the
pre synaptic cell by exocytosis.
After that they get in the synaptic
cleft.
9
Receptor Molecules In Synapses
After that neurotransmitters
interact with the next cell. This way
the nervous impulse is transmitted
between 2 neurons.
10
GABBA
11
Glutamate
Inhibitory Responses Excitatory Responses
o Exocytose vesicles, spontaneously,
merge with the pre synaptic
membrane, propagating a nervous
impulse to the next neuron.
12
o Substance that alters
nervous impulse
transmission.
13
Adenosine
o It is a secreted protein
in humans.
o Encourage the growth
and differentiation of
new neurons.
o BDNF itself is
important for long-
term memory.
14
o It plays an important role
in the consolidation of
information from short to
long-term memory.
15
The goal of this experiment was to address weather BDNF
effects on mIPSC frequency are dependent on A2ARs
activation.
16
Patch Clamp Method
Loose Patch
Cell Atatched
Whole Cell
Outside Out
Patch
Inside Out Patch
18
19
o Removal of the
brain
o Dissection of the
hippocampus
o Acute brain slices
technique
o 300µm slices
o 2 to 4 weeks old
o Anesthetization
o Decapitation
Vibratom Hipppocampal Slice
20
Visibility in
different
plans
Triangularly shaped
Not too bright
21
Videos youtube
22
http://www.youtube.com/watch?v=xwqFdL_ndMo http://www.youtube.com/watch?v=5ZgiacYwkyY
23
Whole Cell Patch Clamp 24
Miniature events of the inhibitory response
25
Adenosine A2A receptor agonist
Adenosine Activates Track B
receptors Potentiates
Effects of BDNF
26
10 minutes of control time - with TTX, KYN, and SCH
40 minutes of data registration of the inhibitory response
10 min || 40 min
TTX, KYN,
SCH (100 nM)
BDNF 10
(ng/mL)
27
28
Inverted Light Microscope
29
Faraday´s Cage
30
Air Table
31
Micromanipulator
32
Recording Electrode
33
34
0 20 40
0
50
100
150
Time (min)
mIP
SC
fre
qu
en
cy
(% b
ase
line
)
BDNF 10 ng/mL
SCH 100 nM
0 20 40
0
50
100
150
Time (min)
mIP
SC
am
plit
ud
e
(% b
ase
line
)
BDNF 10 ng/mL
SCH 100 nM
35
o Adition of SCH prevents the effects of BDNF;
o Results from a single experimente;
o Plenty of room for Biomedical Engeneering to
improve essencial equipement.
36
37
1. http://www.bem.fi/book/04/04x/0427x.htm SLIDE 19
2. http://www.leicabiosystems.com/specimen-preparation/sectioning/vibrating-blade-
microtomes/details/product/leica-vt1000-a/ SLIDE 21
3. http://www.physiologie.uni-freiburg.de/research-techniques/electrophysiology
SLIDE 22
4. http://www.physiologie.uni-freiburg.de/research-techniques/electrophysiology
SLIDE 25