While an increase in the free intracellular calcium concentration promotes exocytosis in most secretory cells it suppresses renin release from juxtaglomerular cells (JG cells). We speculated that this inhibitor effect of calcium might be mediated by a suppression of cAMP formation, since cAMP is the main intracellular stimulator of renin release. In primary cultures of JG cells, calcium-dependent inhibitors of renin release (angiotensinII, endothelin-1, thapsigargin) suppressed renin secretion, which was paralleled by decreases in intracellular cAMP levels [cAMP]. When [cAMP] was clamped by membrane permeable cAMP derivates, renin release was not suppressed by any of the calcium liberators. Additionally, calcium liberators suppressed cAMP levels and renin release in isoproterenol or forskolin pretreated As4.1 cells, a renin-producing cell line that expresses type 5 and 6 adenylate cyclases (AC5, AC6), that are both inhibited by calcium. Indeed, the calcium-dependent inhibition of intracellular cAMP levels and renin release was prevented by siRNA-mediated knockdown of AC5 and/or AC6 expression. Finally, in isolated perfused mouse kidneys, angiotensin II completely inhibited the stimulation of renin secretion induced by adenylate cyclase activation (isoproterenol) but not by membrane permeable cAMP analogues, supporting the conclusion that the suppressive effect of calcium liberators on renin release is mediated by inhibition of adenylate cyclase activity.