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J Physiol. 1991 Apr;435:275-93.

Patch clamp analysis of excitatory synaptic currents in granule cells of rat hippocampus.

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Max-Planck-Institut für biophysikalische Chemie, Göttingen, Germany.


1. Excitatory postsynaptic potentials (EPSPs) and their underlying currents (EPSCs) were recorded from dentate granule cells in thin hippocampal slices of rats using the tight-seal whole-cell recording technique. 2. At resting membrane potentials (ca -60 to -70 mV), the EPSCs clearly consisted of a dominant fast and a smaller slow component. The slow EPSC component markedly increased with depolarization. This resulted in a region of negative slope conductance (between -50 and -30 mV) in the peak current-voltage (I-V) relation of the dual-component EPSC in most neurones. The EPSCs reversed entirely at -1.2 +/- 2.8 mV (n = 15). 3. Using selective antagonists of N-methyl-D-aspartate (NMDA) and non-NMDA excitatory amino acid receptors, two pharmacologically distinct components of the natural EPSCs were isolated. The non-NMDA EPSCs displayed a linear I-V relation. Their rise times (0.5-1.9 ms) were independent of membrane voltage but seemed to depend critically on the precise dendritic location of the synapse. Their decay was approximated by a single exponential with a time constant ranging from 3 to 9 ms. The time course of these EPSCs was independent of changes in extracellular Mg2+. 4. The NMDA EPSCs displayed a non-linear I-V relation. At resting membrane potentials their peak amplitudes were 20 pA and increased steadily with depolarization to -30 mV. At membrane voltages positive to -30 mV the peak I-V relation was linear. The rise times of NMDA EPSCs ranged from 4 to 9 ms and were insensitive to membrane voltage. 5. The NMDA EPSCs decayed biexponentially. Both time constants, tau f and tau s, increased with depolarization in an exponential manner, tau s being more voltage dependent than tau f. Lowering extracellular Mg2+ slightly reduced both rate constants but did not completely abolish their voltage sensitivity. 6. Bath application of NMDA to outside-out patches from granule cells induced single channel currents of 52 pS in nominally Mg(2+)-free solutions. They displayed a burst-like single-channel activity with clusters of bursts lasting several hundreds of milliseconds. Currents through single NMDA receptor channels reversed around 0 mV. 7. The fractional contributions of NMDA and non-NMDA components to peak currents and synaptic charge transfer were assessed. At resting membrane potential the NMDA EPSC component accounted for 23% of the peak current and for 64% of the synaptic charge transfer. The contribution of the NMDA EPSC component to the synaptic charge transfer strongly increased with small depolarizations from rest.

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