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I need to demonstrate the effects of ethanol on a human brain (or other brain with developed telencephalon) in vitro; ideally the effects on vigilance and/or memory.

Could you please suggest such an experiment?

What method would you suggest to demonstrate the effects of ethanol on NMDA receptors or membrane penetration by ethanol?

Edit in respose to the first comment by @MadScientist:

The point is to empirically demonstrate how ethanol affects the cognitive performance of brain (specifically vigilance and memory) and explain the biochemical nature of such an effect on an observable real example (not using computer simulations). It is aimed at mature layman audience.

The experiment has to be in vitro due to ethical and environmental limitations (well, I am not permitted to intoxicate mature students nor animals).

Demonstrating the effect of ethanol in the role of an NMDAR antagonist would be satisfactory as the chain of implications would be illustrated on a theoretical basis.

I have access to standard laboratory equipment on a university level, but the simpler it is, the better. As to the material, the same is true while a fresh brain slice will hopefully be available as well.

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In vivo experiments are much easier to perform for this topic ;-). You'll have to specify what exactly you want to achieve, there's a huge difference between "effect on the brain" and "effect on the NMDS receptor". You'll also have to give us a rough idea on what kind of equipment and material you have access to. –  Mad Scientist Mar 29 '12 at 14:23
@MadScientist I have edited my question to give additional details. –  Harold Cavendish Mar 29 '12 at 15:01
Can you patch? Do calcium imaging maybe? –  nico Mar 29 '12 at 19:47
@nico I have not done it before. Could you perhaps suggest an exact methodology, please? –  Harold Cavendish Apr 3 '12 at 22:02
@Harrold Cavendish: if you haven't done it before, forget about patching, it would be very technically challenging. I will try to write an answer later today, now I'm late for work!!! :D –  nico Apr 4 '12 at 6:30
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up vote 6 down vote accepted

The main problem with what you are asking is that you want to show effects on vigilance and memory in vitro. That is just not possible: if you want vigilance and memory you need a live animal, there is no way around it.

Next point: you have an audience of non scientists, so you can lose them very quickly if you start speaking about NMDA, LTP or similar things without a clear explanation, so start with that: get a nice review (Pubmed is a good start, as always), and give a simple theoretical explanation of one of the mechanisms involved. It is then important to clearly explain the experiment you will perform, and what you are expecting to see in case there is an effect.

Unfortunately the best way to see effects on NMDA receptors would be electrophysiology. This is not trivial and, if you have never done it, I would not suggest going that way. However, if you have access to an electrophysiology setup you will most likely have access to someone who uses it, and that could make the experiment for you.

I am not an expert on this topic, but a quick research on Pubmed seems to indicate that alcohol can, for instance, reduce LTP in hippocampal neuron. Maybe what you could do is having someone patch hippocampal neurons and show how an LTP protocol is used. You don't necessary have to show them the whole thing (I doubt anyone in a right state of mind would bare to stay until the whole experiment is performed in control condition first, and alcohol later...).

Also, the results of these experiments are often non obvious to analyse, so you should have some data prepared and analysed earlier and quickly show how analysis is performed, before showing the final results.

Calcium imaging experiments could also be used and would probably give results that are easier to understand for a laymen audience.

See for instance Fig. 1 or Fig. 5 of this paper (just the first one I found, there surely are other): Ethanol alters calcium signaling in axonal growth cones.

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+100 for noting the requirement of a live animal and how non-trivial the proposed experiments are –  yamad Apr 5 '12 at 17:51
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