I am measuring the effect of a drug on E/I ratio in lateral habenula neurons but I am running into the issue of generating action potentials in the excitatory components. I am using whole-cell patch clamp, AP-V and stim is set so that I am able to generate an evoked eEPSC (amplitude ~ -400) and normal eIPSC before drug treatment. But ~ 1/2 my cells start generating what may be action potentials following drug treatment to the same stimulus with amplitudes ~ -2500 to -3000. Is it possible to have this high of eEPSC or is really action potentials? Would you suggest starting eEPSC ampltiudes lower ~ -200 to see if I can generate more reasonable changes. I don't think E/I ratio of +15 or +30 to even be real. especially if its looks more like an AP... Thank you for any input.
Yes! The currents should be "action potential currents" which correspond with the generation of action potentials identified in current-clamp configuration.
Rather than relying on amplitude, kinetics should be the clue to differentiate between the signals. A recurent AP signal will have faster kinetics (especially during the relaxation) than an ev-EPSC.
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