Catecholamines stimulate renin release by activation the ß-adrenoceptor-cAMP pathway. Recently, it was shown in cultured renin producing cells that the calcium-inhibited adenylyl cyclases AC5 and AC6 play important roles in the control of renin exocytosis. To investigate whether and to what extent AC5 and AC6 are involved in the stimulation of renin secretion in vivo, we performed experiments in AC5 and AC6 knockout mice. Male AC5-/- mice show normal plasma renin concentration (PRC) under control conditions. In contrast, PRC is stimulated in male AC6-/- mice compared to AC6+/+ together with enhanced diuresis and natriuresis and reduced arterial blood pressure. A single injection of the ß-adrenoreceptor agonist isoproterenol markedly stimulated PRC in both knockout strains. However, the stimulation was significantly attenuated in AC5-/- (-20%) and AC6-/- (-63%) compared to their respective wildtypes. Similar results were obtained in the isolated perfused kidney model. Isoproterenol, prostaglandin E2 and pituitary adenylyl cyclase-activating polypeptide (PACAP), that all stimulate renin release via the cAMP pathway, enhanced renin secretion rates (RSR) in all genotypes. However, the stimulation of RSR in response to these drugs/hormones was reduced in AC5-/- (-31 to 46% vs. AC5+/+). This attenuation was even more pronounced in AC6-/- in which RSR was reduced by 51 to 98% compared to AC6+/+. Taken together, our data indicate that both AC5 and AC6 are involved in the stimulation of renin release in response to catecholamines, PGE2 and PACAP, and AC6 is the dominant isoform in this process.