Single Ethanol Withdrawal Regulates Extrasynaptic δ-GABA_A Receptors Via PKCδ Activation

Alcohol (ethanol, EtOH) is one of the most widely abused drugs with profound effects on brain function and behavior. GABA_A receptors (GABA_ARs) are one of the major targets for EtOH in the brain. Temporary plastic changes in GABA_ARs after withdrawal from a single EtOH exposure occur both in vivo and in vitro, which may be the basis for chronic EtOH addiction, tolerance and withdrawal symptoms. Extrasynaptic δ-GABA_AR endocytosis is implicated in EtOH-induced GABA_AR plasticity, but the mechanisms by which the relative abundance and localization of specific GABA_ARs are altered by EtOH exposure and withdrawal remain unclear. In this study, we investigated the mechanisms underlying rapid regulation of extrasynaptic δ-GABA_AR by a single EtOH withdrawal in cultured rat hippocampal neurons. Thirty-minutes EtOH (60 mM) exposure increased extrasynaptic tonic current (I_tonic) amplitude without affecting synaptic GABA_AR function in neurons. In contrast, at 30 min after withdrawal, I_tonic amplitude and responsiveness to acute EtOH were both reduced. Similar results occurred in neurons with okadaic acid (OA) or phorbol 12,13-dibutyrate (PDBu) exposure. Protein kinase C (PKC) inhibition prevented the reduction of I_tonic amplitude and the tolerance to acute EtOH, as well as the reduction of GABA_AR-δ subunit abundance induced by a single EtOH withdrawal. Moreover, EtOH withdrawal selectively increased PKCδ level, whereas PKCδ inhibition specifically rescued the EtOH-induced alterations in GABA_AR-δ subunit level and δ-GABA_AR function. Together, we provided strong evidence for the important roles of PKCδ in the rapid regulation of extrasynaptic δ-GABA_AR induced by a single EtOH withdrawal.