In the rat hippocampus, 5-hydroxytryptamine (5-HT)1A receptors couple to two independent effector mechanisms, the inhibition of adenylyl cyclase activity and the opening of a K+ channel. In the dorsal raphe, 5-HT1A receptors also open K+ channels; however, coupling to adenylyl cyclase has not been demonstrated. In this study, the selective 5-HT1A agonists (+/-)- 8-hydroxy-2-(di-n-propylamino)tetralin, (R+)-8-hydroxy-2-(di-n-propylamino)tetralin and dipropyl-5-carboxamidotryptamine, did not inhibit forskolin-stimulated adenylyl cyclase (FSAC) activity in raphe region homogenates, although these drugs were efficacious in hippocampal homogenates. Other 5-HT1A agonists, NAN-190, BMY-7378, buspirone and gepirone, were also ineffective in raphe region homogenates. Estrogen-treatment of ovariectomized female rats, which is known to enhance 5-HT1A-mediated inhibition of FSAC in the hippocampus, did not promote the action of 5-HT1A agonists. Nor did activation of 5-HT1A receptors stimulate basal adenylyl cyclase activity in raphe homogenates as it does in the hippocampus. FSAC activity was inhibited in raphe region homogenates by activation of adenosine A1 or gamma-aminobutyric acidB receptors or by direct activation of the inhibitor G-protein, Gi, with guanyl-5'-6'-imidodiphosphate, indicating that the raphe homogenates have the biochemical machinery for inhibition of FSAC. High affinity binding studies showed that, in raphe homogenates, 5-HT1A receptors were expressed at a density comparable to that of adenosine A1 receptors and that they were coupled to G-proteins. It should be noted that our failure to observe 5-HT1A-mediated inhibition of adenylyl cyclase in the raphe does not prove that such coupling does not exist. However, a lack of 5-HT1A receptor coupling to adenylyl cyclase in the raphe would support contentions that coupling of the 5-HT1A receptor to adenylyl cyclase may be independent of its coupling to the K+ channel and that there may be distinct differences between pre- and postsynaptic 5-HT1A receptor systems.