G-protein pattern and adenylyl cyclase activity in the brain of rats after long-term ethanol.

Previous studies have described changes in levels of GTP binding proteins (G-proteins) following exposure of rodents to ethanol that did not correlate with the altered activation of the transmembrane signaling pathway. Possible reasons for these inconsistencies were taken into account in the present study by measuring the levels of four different G-protein subunits (G(alpha s), G(alpha i1/2), G(alpha o), Gbetagamma) in six brain regions. Rats were exposed to ethanol for 4 weeks (forced intake of ethanol liquid diet) and 40 weeks (free-choice ethanol). G-protein levels and activation of adenylyl cyclase (AC) were measured on day 1, day 8, and day 28 after withdrawal. When there were changes in the G-protein levels at all, increases were observed mostly in brain regions from rats with the 40-week exposure and decreases in regions from rats with the 4-week exposure that consumed a higher amount of ethanol per day. In some regions the changes had not normalized by day 28 in the 40-week ethanol group whereas in the 4-week ethanol group changes were observed only at day 1 and day 8. Activation of AC was disturbed in the 4-week ethanol group. Reduced activation was detected in membranes of the cerebral cortex, whereas increased activation was observed in the cerebellum, hypothalamus, pons, and striatum. Addition of ethanol (100 mM) to the tissue homogenate facilitated the stimulating action of Gpp(NH)p only in the hippocampus, cerebellum, and striatum. This in vitro action of ethanol was not affected by the long-term ethanol exposure. Activation of AC in the 40-week ethanol group was reduced in the cerebral cortex, pons, and striatum and increased in the cerebellum and hypothalamus if changes occurred at all. The findings support the contention that changes of the transmembrane signaling pathway in ethanol-exposed rats depend on the brain region and on the mode of application. Furthermore, a clear dissociation was observed between changes of the activation of the adenylyl cyclase and the changes in the levels of G-proteins.