Cyclic AMP-induced desensitization of G-protein-regulated phospholipase C in turkey erythrocyte membranes.

The interaction of the cyclic AMP and inositol lipid signalling systems was studied in turkey erythrocytes. Elevation of intracellular cyclic AMP concentrations by pretreatment of the cells with forskolin or 8-Br-cAMP resulted in a marked decrease in responsiveness of phospholipase C to G-protein activators in membranes prepared from treated cells. Decreases in responsiveness occurred with a t1/2 of approximately 5 min and were reversible after transfer of desensitized cells to drug-free medium. Pretreatment of the cells with forskolin inhibited inositol phosphate formation in a concentration-dependent manner and addition of the phosphodiesterase inhibitor IBMX 93-isobutyl-1-methylxanthine) during pretreatment increased the capacity of forskolin to desensitize phospholipase C activity. IBMX also produced a similar potentiation of forskolin-stimulated accumulation of cyclic AMP in turkey erythrocytes. Isoproterenol pretreatment of the cells induced, like forskolin, partial inhibition of inositol phosphate generation in response to G-protein activators and to P2y purinoceptor and beta-adrenoceptor agonists. The capacity of isoproterenol to induce desensitization of phospholipase C activity also was increased by the presence of IBMX during pretreatment of the cells. H8 (N-[2-(methylamino)ethyl]-5-isoquinoline-sulfonamide), an inhibitor of cyclic AMP-regulated protein kinase, completely prevented forskolin-induced desensitization but only partially blocked isoproterenol-induced desensitization. These results indicate that the cyclic AMP signalling cascade has a major inhibitory influence on receptor- and G-protein-activated inositol lipid signaling.