Journal
NEUROSCIENCE
Volume 162, Issue 4, Pages 1187-1191Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.neuroscience.2009.05.031
Keywords
subunit composition; furosemide; zinc; tonic inhibition; ethanol sensitivity
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GABA-activated chloride currents were studied in cerebellar granule cells put in culture from neonatal rats. As previously described, 10 mu M GABA perfusion of these cells recorded by whole cell patch-clamp elicits chloride currents displaying a peak and a steady-state component. The two components were studied in the presence of 1 mM furosemide, 1 mu M Zn2+ and a combination of the two in order to evaluate the contribution of the different types of GABA(A) receptors. Furosemide inhibits alpha(6) containing receptors whereas low levels of Zn2+ specifically block incomplete GABA(A) receptors made up of a and beta subunits only. The results show that the peak component involves the following receptors: alpha(x) beta(y), 25%; alpha(1) beta(y) gamma(2), 45%; alpha(6) beta(y) gamma(2) plus alpha(1) alpha(6) beta(y) gamma(2) 30%. The steady state component is made up by alpha(x) beta(y), 38%; alpha(1) beta(y) delta, 62%. Ethanol at relatively high concentration, 100 mM, slows further down the desensitization of alpha(1) beta(y) delta receptors. The results indicate that the relative insensitivity to ethanol of GABA(A) receptors of neonatal cerebellar granule cells in culture is due to the absence of mature alpha(6) beta(y) delta receptors, a major receptor brand involved in tonic inhibition. (C) 2009 IBRO. Published by Elsevier Ltd. All rights reserved.
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