This study examined adenylyl cyclase (AC) signal transduction in alcohol-sensitive brain regions of rats selectively bred for high (HAD) and low (LAD) alcohol drinking and correlated these findings with differences in sensitivity and tolerance to alcohol-induced sedation found within these lines. LAD rats were more sensitive to the sedative effects of alcohol than were HAD rats as evidenced by a shorter latency to lose the righting response (RR) after a single alcohol challenge. When time to recover the RR was compared after each of two alcohol challenges, HAD rats recovered the RR more rapidly following the second challenge compared to the first, indicating that the HAD rats rapidly developed tolerance to the sedative effects of alcohol. Tolerance did not develop in rats of the LAD line. Two months after completion of behavioral testing, adenylyl cyclase (AC) signal transduction was examined in alcohol-sensitive brain regions of rats from both lines. Immunoblot analyses indicated that LAD rats had greater Gs alpha expression in the frontal cortex (FC) and hippocampus (HIP) compared to HAD rats. Rats with the highest HIP and FC Gs alpha levels were more rapidly affected by the sedative properties of alcohol than were rats with lower Gs alpha levels. G protein expression and AC activity in the FC, HIP, cerebellum (CERE), and nucleus accumbens (ACB) were also correlated with sensitivity to the sedative properties of alcohol and with the rapid development of tolerance to this alcohol effect. The results suggest that sensitivity and tolerance to alcohol-induced sedation may be mediated in part through AC signal transduction.