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GABAA-mediated miniature IPSCs (mIPSCs) were recorded from layer V pyramidal neurons of the visual cortex using whole-cell patch-clamp recording in rat brain slices. At room temperature, the benzodiazepine site agonist zolpidem enhanced both the amplitude (to 138 ± 26% of control value at 10 μm) and the duration (163 ± 14%) of mIPSCs. The enhancement of mIPSC amplitude was not caused by an increase of the single-channel conductance of the postsynaptic receptors, as determined by peak-scaled non-stationary fluctuation analysis of mIPSCs. The effect of zolpidem on fast, synaptic-like (1 msec duration) applications of GABA to outside-out patches was also investigated. The EC50 for fast GABA applications was 310 μm. In patches, zolpidem enhanced the amplitude of currents elicited by subsaturating GABA applications (100–300 μm) but not by saturating applications (10 mm). The increase of mIPSC amplitude by zolpidem provides evidence that the GABAA receptors are not saturated during miniature synaptic transmission in the recorded cells. By comparing the facilitation induced by 1 μm zolpidem on outside-out patches and mIPSCs, we estimated the concentration of GABA seen by the postsynaptic GABAA receptors to be ∼300 μm after single vesicle release. We have estimated a similar degree of receptor occupancy at room and physiological temperature. However, at 35°C, zolpidem did not enhance the amplitude of mIPSCs or of subsaturating GABA applications on patches, implying that, in these neurons, zolpidem cannot be used to probe the degree of receptor occupancy at physiological temperature.

Effect of Zolpidem on Miniature IPSCs and Occupancy of Postsynaptic GABAAReceptors in Central Synapses

Effect of Zolpidem on Miniature IPSCs and Occupancy of Postsynaptic GABA_AReceptors in Central Synapses