The role of G proteins in the activity and ethanol modulation of glycine-induced currents in rat neurons freshly isolated from the ventral tegmental area.

In freshly isolated neurons of the ventral tegmental area of young rats, we first examined the role of G proteins in the functional modulation of the glycine receptor (GlyR). GTP-gamma-S [guanosine-5'-0-(2-thiotriphosphate)] (2 mM) or GDP-beta-S [guanosine 5'-0-(2-thiodiphosphate)] (2 mM) was added to the pipette solution of whole-cell recordings to regulate G protein activities. GTP-gamma-S enhanced the amplitude of glycine-induced current (I(Gly)), suggesting modulation of GlyRs via a G protein-coupled pathway. GDP-beta-S suppressed I(Gly), suggesting that basal G protein activity positively modulates the GlyRs. We next examined effects of G proteins in ethanol potentiation of GlyR function. Activation of G proteins with 2 mM GTP-gamma-S attenuated, but did not eliminate, ethanol-induced potentiation of I(Gly). These results suggest that GTP-gamma-S and ethanol share the same pathway of activating GlyRs. When G proteins are maximally activated by GTP-gamma-S, the action of ethanol was partially occluded. When 2 mM GDP-beta-S was added in pipette solution, ethanol-induced potentiation of I(Gly) was significantly attenuated, suggesting that GDP-beta-S partially blocked the action of ethanol. However, the inability of GTP-gamma-S (or GDP-beta-S) to eliminate completely the potentiating effect of ethanol indicates that some other factors, in addition to G proteins, may also contribute to the action of ethanol on GlyRs.