Agmatine, decarboxylated arginine, is produced in the kidney and can increase nephron and kidney filtration rate via renal vasodilatation and increases in plasma flow. This increase in filtration rate after agmatine is prevented by administration of nitric oxide synthase (NOS) inhibitors. In endothelial cells agmatine stimulated nitrite production is accompanied by induction of cytosolic calcium. NOS activity requires calcium for activation; however the source of this calcium remains unknown. Here we demonstrate ryanodine receptor calcium activated calcium release (RyR) channels in the kidney cortex, and evaluate if RyR contributes to the agmatine response. Agmatine microperfused into Bowman s space reversibly increases nephron filtration rate (SNGFR) by ~30%. Cyclic ADP-ribose (cADPR) regulates RyR channel activity. Concurrent infusion of agmatine with the cADPR blocker, 8-bromo-cADPR (2µM), prevents the increase in filtration rate. Furthermore, direct activation of the RyR channel with ryanodine at agonist concentrations (5 µM) increases SNGFR, and like agmatine, this increase is prevented by administration of L-NMMA, a non-selective NOS blocker. We demonstrate that agmatine does not elicit ADPR cyclase activity in vascular smooth muscle membranes and does not directly affect RyR calcium channel responses using sea urchin egg homogenates. These results imply interplay between endothelial cell cADPR/RyR/Ca++/NO and the cADPR/RyR/Ca++ pathways in vascular smooth muscle cells in arterioles in the regulation of kidney filtration rate. In conclusion, we show that agmatine-induced effects require activation of cADPR and RyR calcium release channels for NO generation, vasodilation and increased filtration rate.