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Background and Purpose: The human 5-hydroxytryptamine(7) (h5-HT(7)) receptor is G(s) -coupled and stimulates the production of the intracellular signalling molecule cAMP. Previously, we reported a novel property of the h5-HT(7) receptor: pseudo-irreversible antagonists irreversibly inhibit forskolin-stimulated (non-receptor-mediated) cAMP production. Herein, we sought to determine if competitive antagonists also affect forskolin-stimulated activity and if this effect is common among other G(s) -coupled receptors.Experimental Approach: Recombinant cell lines expressing h5-HT(7) receptors or other receptors of interest were briefly exposed to antagonists; cAMP production was then stimulated by forskolin and quantified by an immunocompetitive assay.Key Results: In human embryonic kidney 293 cells stably expressing h5-HT(7) receptors, all competitive antagonists inhibited nearly 100% of forskolin-stimulated cAMP production. This effect was insensitive to pertussis toxin, that is, not G(i/o) -mediated. Potency to inhibit forskolin-stimulated activity strongly correlated with h5-HT(7) binding affinity (r(2) = 0.91), indicating that the antagonists acted through h5-HT(7) receptors to inhibit forskolin. Potency and maximal effects of clozapine, a prototypical competitive h5-HT(7) antagonist, were unaffected by varying forskolin concentration. Antagonist interaction with h5-HT(6), human β(1), β(2), and β(3) adrenoceptors did not inhibit forskolin's activity.Conclusions and Implications: The inhibition of adenylate cyclase, as measured by forskolin's activity, is an underlying property of antagonist interaction with h5-HT(7) receptors; however, this is not a common property of other G(s) -coupled receptors. This phenomenon may be involved in the roles played by h5-HT(7) receptors in human physiology. Development of h5-HT(7) antagonists that do not elicit this effect would aid in the elucidation of its mechanisms and shed light on its possible physiological relevance. [ABSTRACT FROM AUTHOR]