RT Journal Article SR Electronic T1 Plasticity of GABAA Receptors after Ethanol Pre-Exposure in Cultured Hippocampal Neurons JF Molecular Pharmacology JO Mol Pharmacol FD American Society for Pharmacology and Experimental Therapeutics SP 432 OP 442 DO 10.1124/mol.110.068650 VO 79 IS 3 A1 Yi Shen A1 A. Kerstin Lindemeyer A1 Igor Spigelman A1 Werner Sieghart A1 Richard W. Olsen A1 Jing Liang YR 2011 UL http://molpharm.aspetjournals.org/content/79/3/432.abstract AB Alcohol use causes many physiological changes in brain with behavioral sequelae. We previously observed (J Neurosci 27:12367–12377, 2007) plastic changes in hippocampal slice recordings paralleling behavioral changes in rats treated with a single intoxicating dose of ethanol (EtOH). Here, we were able to reproduce in primary cultured hippocampal neurons many of the effects of in vivo EtOH exposure on GABAA receptors (GABAARs). Cells grown 11 to 15 days in vitro demonstrated GABAAR δ subunit expression and sensitivity to enhancement by short-term exposure to EtOH (60 mM) of GABAAR-mediated tonic current (Itonic) using whole-cell patch-clamp techniques. EtOH gave virtually no enhancement of mIPSCs. Cells pre-exposed to EtOH (60 mM) for 30 min showed, 1 h after EtOH withdrawal, a 50% decrease in basal Itonic magnitude and tolerance to short-term EtOH enhancement of Itonic, followed by reduced basal mIPSC area at 4 h. At 24 h, we saw considerable recovery in mIPSC area and significant potentiation by short-term EtOH; in addition, GABAAR currents exhibited reduced enhancement by benzodiazepines. These changes paralleled significant decreases in cell-surface expression of normally extrasynaptic δ and α4 GABAAR subunits as early as 20 min after EtOH exposure and reduced α5-containing GABAARs at 1 h, followed by a larger reduction of normally synaptic α1 subunit at 4 h, and then by increases in α4γ2-containing cell-surface receptors by 24 h. Measuring internalization of biotinylated GABAARs, we showed for the first time that the EtOH-induced loss of Itonic and cell-surface δ/α4 20 min after withdrawal results from increased receptor endocytosis rather than decreased exocytosis.