Journal
NEUROPHARMACOLOGY
Volume 39, Issue 12, Pages 2408-2417Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/S0028-3908(00)00095-2
Keywords
gamma-aminobutyric acid; GABA(A)-antagonists; electrophysiology; synchronisation; calcium; bicuculline; gabazine; multiple microelectrode
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The present study was performed to investigate the effects of depleting intracellular Ca2+ stores on bicuculline- or gabazine-induced epileptiform excitability. Studies were performed on monolayer rat hippocampal neuronal networks utilising a system that allowed simultaneous multiple extracellular single-unit recordings of neuronal activity. Hippocampal neuronal networks were prepared from enzymatically dissociated hippocampi from 18-day-old fetal Wistar rats. The cells were cultured in Neurobasal medium with B27 serum-free supplements directly onto the surface of planar multiple microelectrode arrays with a central recording array of 64 (4x16) indium-tin thin-film recording electrodes. All cells recorded at 21 days-in-vitro exhibited spontaneous discharge activity with firing rates between 0.3-30.7 Hz. gamma-aminobutyric acid (GABA) produced a concentration-dependent decrease in firings (EC50=9.1 mu M) which could be blocked by pre-application of bicuculline methobromide (10 mu M). Addition of the GABA(A)-receptor antagonists gabazine (10 mu M) or bicuculline (10 mu M) resulted in the rapid generation of synchronised bursting within all the cells recorded. Bicuculline exhibited heterogeneity of action on firing rate, whereas gabazine always increased firing. Pre-incubation with thapsigargin, which depletes intracellular calcium stores, resulted in a decrease in the amount of neuronal excitation produced by bicuculline, but not by gabazine, suggesting that bicuculline-induced neuronal excitation requires release of Ca2+ from intracellular stores. (C) 2000 Elsevier Science Ltd. All rights reserved.
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