Hi all,
I am culturing primary hippocampal cells from P0 mice and would like to measure whether synaptic plasticity is altered after treatment with different compounds, using whole-cell patch clamp.
What are your favourite techniques/protocols for plasticity measurements in dissociated cell culture using whole-cell patch clamp or other techniques?
We have the options of spike-timing dependent plasticity, chemically-induced LTP, etc.
Thanks in advance,
Hannah
Dear Hannah,
I would not recommend dissociated neuronal cultures for studying synaptic plasticity, as pretty much most synapses are removed/damaged during the process. Freshly cut brain slices preserving some pre- and post synaptic components and glial-neuronal architecture commonly used by many researchers studying neuronal plasticity, and there are many previous methodological papers describing details and protocols.
best wishes,
Refik
Thank you for your responses Nicolas Paradis-Isler - they are much appreciated and I will look into those methods of chemical LTP/LTD.
Hi Annah,
in order to induce plasticity in neuronal cultures you can pursue 2 different streets. The first one is the chemical induction using a modified external solution supplemented with glycine at saturating concentration for 3 minutes. The second approach could be the dual patch where you will identify 2 connected neurons and after that you stimulate the presynapse in order to potentiate the post. However, the first method is easier than the second one, and it works pretty well.
Good luck for your experiments
Luca
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