A method for recording miniature inhibitory postsynaptic currents in the central nervous system suitable for quantal analysis.
Quantal analysis of transmitter release is useful in examining presynaptic mechanisms involved in synaptic transmission. However, in central neurons, the presence of multiple synapses makes it difficult to use the traditional quantal analysis, developed for the neuromuscular transmission. We developed a method to minimize these difficulties. Experiments were performed, using the whole-cell patch-clamp recording technique, on rat CA1 pyramidal neurons in a hippocampal slice preparation. When the stratum radiatum was stimulated, mixed current signals including, miniature inhibitory postsynaptic currents (mIPSCs), miniature excitatory postsynaptic currents (mEPSCs), evoked inhibitory postsynaptic currents (eIPSCs) and evoked excitatory postsynaptic currents (eEPSCs), could be observed in CA1 pyramidal cells while slices were superfused with the normal, Na(+)-containing, medium. The mIPSCs could be blocked by bicuculline (10 microm). mEPSCs, eEPSCs and eIPSCs could not be observed when the Na(+)-containing perfusion medium was replaced by a Na(+)-free medium but reappeared when the Na(+)-containing medium was re-introduced. When a polarizing electrode was placed near the recorded neuron, while slices were superfused with the Na(+)-free medium, and depolarizing rectangular current pulses of different magnitudes were applied, the number of mIPSCs increased with increasing amount of the current. Amplitudes of the mIPSCs showed a Gaussian distribution and the coefficient of variation was small. These observations indicate that a combination of the Na(+)-free superfusing medium and local depolarizations with a polarizing electrode is useful for recording mIPSCs from a localized area of the recorded neuron and for quantal analysis.
Singh
G
G
Neuroscience Research Laboratory, Department of Anesthesiology, Pharmacology and Therapeutics, Faculty of Medicine, The University of British Columbia, Vancouver, Canada V6T 1Z3.
Xu
J-Y
JY
Sastry
B R
BR
D000818:Animals; D000831:Animals, Newborn; D001640:Bicuculline / Q000494:pharmacology; D004307:Dose-Response Relationship, Radiation; D004558:Electric Stimulation / Q000379:methods*; D019706:Excitatory Postsynaptic Potentials / Q000187:drug effects / Q000502:physiology; D018756:GABA Antagonists / Q000494:pharmacology; D006624:Hippocampus / Q000166:cytology*; D066298:In Vitro Techniques; D053444:Inhibitory Postsynaptic Potentials / Q000187:drug effects / Q000502:physiology*; D008297:Male; D016011:Normal Distribution; D018408:Patch-Clamp Techniques / Q000379:methods; D017966:Pyramidal Cells / Q000502:physiology*; D051381:Rats; D017208:Rats, Wistar; D012964:Sodium / Q000378:metabolism
D018756:GABA Antagonists; D012964:Sodium; D001640:Bicuculline