Hi everyone,
I am trying to record EPSCs on medium spiny neurons from corticostriatal afferents in the dorsal striatum. The problem is that most neurons I patch show very small EPSCs or no EPSCs at all.
Would you give me some tips on recording EPSCs?
Specifically:
- I am cutting mouse (8-12 weeks old) striatum slices with NMDG-ACSF.
- Although I am more or less succesful obtaining whole-cell recordings from MSNs, those almost never show any EPSC.
- I cut 250 µm parasagittal slices, I place the stim electrode on the corpus callosum and stimulate with 100 µs pulses of 0.1 - 1 mA.
- Important: I intend to use K-gluconate intracellular solution for both CC and VC recordings, because first I want to identify the neuron as MSN (based on firing properties) and then elicit EPSCs in voltage clamp (-70 mV).
Is there any advice you could give me on obtaining corticostriatal EPSCs on dorsal striatum MSNs?
Thanks a lot for your attention!
Dear Diego,
As a general rule in electrophysiology, if you want to record EPSCs in voltage-clamp, you will need to use cesium-based internal solution. Cesium will block many K+ channels and increase the synaptic EPSC amplitude. If you use potassium you will record small amplitude EPSCs as K channels will be active at holding voltages.
For example see the following technical papers for internal (pipette) solutions that I use.
Article Semi-loose seal Neurobiotin electroporation for combined str...
Article Neurobiotin Electroporation for Combined Structural and Func...
best wishes,
Refik
Hi Diego,
I don't know which Ca++ and Mg++ concentrations are you using in your aCSF, but you can try to increase a little bit Ca++ in order to increase glutamate release.
Good luck
Luca
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