Renin is synthesized and released from juxtaglomerular (JG) cells. Adenosine inhibits renin release via an adenosine A1 receptor (A₁R), calcium-mediated pathway. How this occurs is unknown. In cardiomyocytes, adenosine increases intracellular calcium via transient receptor potential canonical (TRPC) channels. We hypothesized adenosine inhibits renin release via A₁R activation, opening TRPC channels. However, higher concentrations of adenosine may stimulate renin release through A₂R activation. Using primary cultures of isolated mouse JG cells, Immunolabeling demonstrated renin and A₁R in JG cells, but not A₂R subtypes, though RT-PCR indicated mRNA of both A_2AR and A_2BR. Incubating JG cells with increasing concentrations of adenosine decreased renin release. Different concentrations of the adenosine receptor agonist NECA didn't change renin. Activating A₁R with 0.5µM of CHA decreased basal renin release from 0.22±0.05 to 0.14±0.03 µg AngI/ml/mg prot (p<0.03), and higher concentrations also inhibited renin. Reducing extracellular calcium with EGTA increased renin release (0.35±0.08 µgAngI/ml/mg prot (p<0.01), and blocked renin inhibition by CHA (0.28±0.06 ugAngI/ml/mg prot p<0. 005 vs. CHA alone). The intracellular calcium chelator BAPTA-AM increased renin release by 55%, and blocked the inhibitory effect of CHA. Repeating these experiments in JG cells from A₁R knockout mice using CHA or NECA demonstrated no effect on renin release. However, RT-PCR showed mRNA from TRPC 3 and 6 in isolated JG cells. Adding the TRPC blocker SKF-96365 reversed CHA-mediated inhibition of renin release. Thus, A₁R activation results in a calcium-dependent inhibition of renin release via TRPC-mediated calcium entry, but A2 receptors do not regulate renin release.