Modulation of decay kinetics and frequency of GABA A receptor-mediated spontaneous inhibitory postsynaptic currents in hippocampal neurons

Abstract

Inhibitory postsynaptic currents mediated by spontaneous activation of GABA A receptors were studied using whole-cell voltage-clamp recordings in granule cells of the adult rat (postnatal day 60+) dentate gyrus in 400-μm-thick coronal half-brain slices maintained at 34–35°C. The average amplitude of spontaneous inhibitory postsynaptic currents remained constant during a given recording period (i.e. no rundown was noted). The spontaneous currents had an average conductance between 200–400 pS, were mediated by Cl − flux through GABA A receptor/channels since they reversed at the Cl − equilibrium potential and were blocked by bicuculline or picrotoxin. Their mono-exponential decay time-constants (range: 4.2–7.2 ms) were prolonged by midazolam and pentobarbital in a dose-dependent manner. The effect of midazolam was reversed by the benzodiazepine receptor antagonist flumazenil (RO 15–1788) which, by itself, had no effect on the decay time-constant. The decay time-constant was also dependent on membrane voltage and on temperature. A 132-mV change in membrane potential produced an e-fold prolongation of the decay while the Q 10 (between 22–37°C) of the decay rate was 2.1. Within a given neuron, the frequency of spontaneous GABAergic events was remarkably constant over long time-periods, though the mean frequency among different cells showed large variability. Spontaneous miniature inhibitory postsynaptic currents also persisted under experimental conditions such as the presence of extracellular tetrodotoxin (1 μM), Cd 2+ (200 μM) or lowered extracellular Ca 2+/elevated Mg 2+, which effectively abolished all stimulus-evoked GABAergic neurotransmission. The frequency of tetrodotoxin-resistant miniature events was increased by elevating extracellular K + concentration and was diminished by the GABA B receptor agonist (−)baclofen only at a dose (50 μM) which was an order of magnitude larger than that required to depress stimulus-evoked responses. These findings are consistent with different mechanisms being responsible for the spontaneous and stimulus-evoked release of GABA from interneuron terminals and also identify pre- and postsynaptic modulatory factors of the endogenous, action-potential-independent, GABAergic neurotransmission as being important determinants of the excitability level of mammalian CNS neurons.