Outward rectification of inhibitory postsynaptic currents in cultured rat hippocampal neurones

Abstract

1. Inhibitory postsynaptic potentials (IPSPs) and currents (IPSCs) were recorded from cultured hippocampal neurones of the embryonic rat at 22 degrees C, using the whole-cell patch-clamp technique with a low-Cl-, 145 mM-potassium gluconate solution in the patch pipette. Individual synaptic events were elicited at low frequency (0.05-0.1 Hz) by stimulating a presynaptic neurone either by direct intracellular current injection, or by applying a brief pulse of L-glutamate. 2. In target neurones voltage clamped at -40 mV, outwardly directed IPSCs of mean amplitude 0.23 nA were recorded. The IPSCs were depressed by the GABA antagonist bicuculline, and reversed polarity between -50 and -80 mV (mean -64 mV), as did current responses to gamma-aminobutyric acid. The IPSPs and IPSCs reversed as a single phase; no bicuculline-resistant 'late' synaptic event was observed. 3. The IPSCs had variable kinetics, with rise times between 1 and 5 ms (mean 2.9 ms) at -40 mV, and slower, monoexponential, decay phases (decay time constant, tau IPSC, 10-40 ms at -40 mV). In some cells, tau IPSC clearly increased with depolarization. 4. The IPSC reversal potential was -64 +/- 9 mV (n = 23) under the experimental conditions used; this suggests that the synaptically activated channels are approximately 25 times more permeable to Cl- than to the gluconate anion. 5. The peak conductance associated with the IPSC showed outward rectification. The synaptic conductance measured at -40 mV was 1.7 times greater than that measured at -100 mV; at -20 mV, synaptic conductance was 2.5 times greater than at -100 mV. This outward rectification can be explained by a constant field model under these experimental conditions of asymmetric Cl- concentrations.