Abstract
BACKGROUND: We tested the hypothesis that central alpha(2A)-adrenergic receptor (alpha(2A)AR) signaling plays a key role in clonidine-ethanol evoked synergistic behavioral impairment. METHODS: Male Sprague-Dawley rats, with intracisternal and jugular vein cannulae implanted 6 days earlier, were tested for drug-induced behavioral impairment. The latter was assessed as the duration of loss of righting reflex (LORR) and rotorod performance every 15 minutes until the rat recovered to the baseline walk criterion (180 seconds). In a separate cohort, c-Fos expression in locus coeruleus (LC) and cerebellum was determined as a marker of neuronal activity following drug treatment. RESULTS: Rats that received clonidine (60 microg/kg, i.v.) followed by ethanol (1 g/kg, i.v.) exhibited synergistic impairment of rotorod performance and LORR. The mixed alpha(2A)AR and I(1)-imidazoline receptor agonist clonidine (30, 60, and 90 microg/kg) synergistically and dose-dependently enhanced behavioral impairment elicited by ethanol (1 g/kg). Possible involvement of I(1)-imidazoline receptors was ruled out because selective I(1)-agonist rilmenidine (300 microg/kg, i.v.) did not cause behavioral impairment alone or enhance ethanol-evoked behavioral impairment. Pharmacological blockade of central alpha(2A)AR (RX821002, 0.3 mg i.c.) abolished the synergy between clonidine and ethanol; the behavioral response caused by the drug combination was similar to that caused by ethanol alone. Conversely, involvement of central alpha(2B)AR in the interaction was ruled out because blockade of central alpha(2B)AR (ARC-239) independently evoked a strong sedative effect. Clonidine (60 microg/kg) or ethanol (1 g/kg) alone increased, but their combination decreased, c-Fos levels in LC, while inconsistent c-Fos responses were observed in cerebellum. CONCLUSIONS: Central alpha(2A)AR, but not I(1)-imidazoline or alpha(2B)AR, signaling is implicated in the synergistic enhancement of ethanol-evoked behavioral impairment by clonidine. Although the mechanism of c-Fos response remains to be investigated, this neurochemical response highlights the LC as a neuroanatomical target for clonidine-ethanol behavioral interaction.