This document discusses an experiment examining the effects of BDNF on miniature inhibitory postsynaptic currents (mIPSCs) and whether these effects are dependent on activation of adenosine A2A receptors (A2ARs). The experiment involved performing whole-cell patch clamp recordings on mouse hippocampal slices to measure mIPSC frequency and amplitude over time under different drug conditions. Results showed that addition of the A2AR agonist SCH 100 nM prevented the effects of BDNF 10 ng/mL on mIPSC frequency and amplitude, suggesting the effects of BDNF are dependent on A2AR activation. The document also provides information on techniques used such as brain slice preparation and patch clamp methodology.

1. Ana Margarida Araújo
Ana Rita Oliveira
Felicia Negru
Filipa Leal
Luís Rita
Professora Ana Sebastião 1

2. Nervous Impulse
Transmission

3. 3

4. 4

5. 5

6. Synapse

7. Electric
Chemical
7

8. Chemical Synapses
Action Potential is transmitted by
special proteins called
neurotransmitters.
8

9. Neurotransmitters
Releasing
Neurotransmitters get out from the
pre synaptic cell by exocytosis.
After that they get in the synaptic
cleft.
9

10. Receptor Molecules In
Synapses
After that neurotransmitters
interact with the next cell. This way
the nervous impulse is transmitted
between 2 neurons.
10

11. GABBA
11
Glutamate
Inhibitory Responses Excitatory Responses

12. o Exocytose vesicles, spontaneously,
merge with the pre synaptic
membrane, propagating a nervous
impulse to the next neuron.
12

13. o Substance that alters
nervous impulse
transmission.
13
Adenosine

14. 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

15. o It plays an important role
in the consolidation of
information from short to
long-term memory.
15

16. The goal of this experiment was to address weather BDNF
effects on mIPSC frequency are dependent on A2ARs
activation.
16

17. Patch Clamp
Method

18. Loose Patch
Cell Atatched
Whole Cell
Outside Out
Patch
Inside Out
Patch
18

19. 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

20. Vibratom Hipppocampal Slice
20

21. Visibility in
different
plans
Triangularly shaped
Not too
bright
21

22. Videos youtube
22

23. http://www.youtube.com/watch?v=xwqFdL_ndMo
http://www.youtube.com/watch?v=5ZgiacYwkyY
23

24. Whole Cell Patch Clamp
24

25. Miniature events of the inhibitory response
25

26. Adenosine A2A receptor agonist
Adenosine Activates
Track B
receptors
Potentiates
Effects of
BDNF
26

27. 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. 28
Inverted Light
Microscope

29. 29
Faraday´s Cage

30. 30
Air Table

31. 31
Micromanipulator

32. 32
Recording Electrode

33. 33

34. 34

35. 0 20 40
0
50
100
150
Time (min)
mIPSCfrequency
(%baseline)
BDNF 10 ng/mL
SCH 100 nM
0 20 40
0
50
100
150
Time (min)
mIPSCamplitude
(%baseline)
BDNF 10 ng/mL
SCH 100 nM
35

36. 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. 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