The human 5-hydroxytryptamine 1A receptor differentially modulates phospholipase C and adenylyl cyclase activities

In this study, we quantitate and compare the ability of the 5-hydroxytryptamine 1A (5-HT1A) receptor to modulate the activities of phospholipase C and adenylyl cyclase as a function of receptor concentration. We used a single clonal cell line permanently expressing the human 5-HT1A receptor, and progressively depleted the receptor concentration using an alkylating antagonist (N8-bromoacetyl-N1-3'-(4-indolyloxy)-2'-hydroxypropyl-Z-1,8-diamin o-p-methane, (+-) Pindobind). For serotonin-induced phospholipase C stimulation, reductions in receptor number result in dose-response curves that shift downward and rightward, reflecting both a decreasing maximal effect as well as an increasing ED50. In contrast, depletion of more than 95% of the receptors has no effect on the maximal inhibition of forskolin-stimulated adenylyl cyclase activity. Moreover, at all receptor concentrations, the amount of serotonin required to produce half-maximal phospholipase C stimulation is several-fold more than that required to produce half-maximal inhibition of adenylyl cyclase activity. We conclude that the 5-HT1A receptor modulates these two pathways differently, and that the overall response to challenge with serotonin, in terms of both phosphatidyl inositol hydrolysis and cyclic AMP production, is dependent upon receptor number.