We have previously shown that pretreatment of A-10 smooth muscle cells (SMC) with angiotensin II (Ang II) attenuated atrial natriuretic peptide (ANP) receptor-C (ANP-C)-mediated inhibition of adenylyl cyclase without altering (125)I-ANP binding. In the present studies, we have investigated the modulation of ANP-C receptor signaling by endothelin-1 (ET-1). Pretreatment of A-10 SMC with ET-1 for 24 h attenuated the expression of ANP-C receptor by about 60% as determined by immunoblotting which was reflected in attenuation of ANP-C-receptor-mediated inhibition of adenylyl cyclase. C-ANP(4-23) [des(Gln(18),Ser(19),Gln(20),Leu(21),Gly(22))ANP(4-23)-NH(2)], a ring-deleted peptide of ANP that interacts specifically with ANP-C receptor, inhibited adenylyl cyclase activity in a concentration-dependent manner with an apparent K(i) of about 1 nM in control cells. The maximal inhibition observed was about 30% which was almost completely attenuated in ET-1-treated cells. In addition, Ang II- and oxotremorine-mediated inhibitions of adenylyl cyclase were also attenuated by ET-1 treatment; however, the expression of Gialpha-2 and Gialpha-3 proteins and not of Gsalpha and Gbeta proteins was augmented by such treatment. The increased expression of Gialpha-2 and Gialpha-3 proteins by ET-1 treatment was inhibited by actinomycin D treatment (RNA synthesis inhibitor). On the other hand, the Gsalpha-mediated effects of some agonists on adenylyl cyclase activity were significantly decreased by ET-1 treatment. These results suggest that ET-1-induced downregulation of ANP-C receptor and not the overexpression of Gi proteins may be responsible for the attenuation of C-ANP(4-23)-mediated inhibition of adenylyl cyclase activity. From these studies it may be suggested that the downregulation of ANP-C receptors by increased levels of endothelin in vivo may be one of the possible mechanisms for the pathophysiology of hypertension.
(c) 2002 Elsevier Science (USA).