Differential effects of epinephrine and norepinephrine on cAMP response and g(i3)alpha protein expression in cultured sympathetic neurons.

The effect of 24-h pretreatment with epinephrine (EPI) or norepinephrine (NE) on alpha(2)- and beta-adrenoceptor agonist-induced, cAMP responses and G(i3)alpha-protein expression was studied in primary cultures of rat superior cervical ganglionic (SCG) neurons. SCG neurons, 10 to 12 days in culture, accumulated cAMP when stimulated with the beta-adrenoceptor agonist isoproterenol and the preferential beta(2)-adrenoceptor antagonist ICI 118,551 blocked this response. Similarly, the preferential alpha(2)-adrenoceptor agonist UK14,304 inhibited forskolin-stimulated cAMP accumulation, implying that cultured SCG neurons possess functional alpha(2)- and beta(2)-adrenoceptors. A 24-h treatment of SCG neurons with EPI or NE induced desensitization of the cAMP response to the beta-adrenoceptor agonist isoproterenol. Simultaneously, EPI treatment increased the maximal inhibitory cAMP response to the alpha(2)-adrenoceptor agonist UK14,304 and NE was without effect. Immunoblotting analyses of G(i3)alpha subunits revealed that 24-h EPI but not NE treatment induces a 3- to 4-fold increase in the expression of G(i3)alpha subunits. Furthermore, EPI-induced up-regulation of alpha-subunit expression can be blocked by the preferential beta(2)-adrenoceptor antagonist ICI 118,551 but not by the preferential beta(1)-adrenoceptor antagonist CGP 20712A. Our results suggest that changes in alpha(2)-adrenoceptor responsiveness induced by EPI may involve activation of beta(2)-adrenoceptors that influence the expression of inhibitory G proteins. Thus, primary cultures of sympathetic neurons by possessing functional alpha(2)- and beta-adrenoceptors may be a suitable model system to study the signaling mechanisms of "cross talk" between these adrenoceptor subtypes, which are known to play a central role in cardiovascular function.