Nucleotides have diverse effects on water and electrolyte reabsorption within the distal tubule of the nephron. As the distal tubule is important in control of renal Mg(2+) balance, we determined the effects of ATP on cellular Mg(2+) uptake in this segment. The effects of ATP on immortalized mouse distal convoluted tubule (MDCT) cells were studied by measuring Mg(2+) uptake with fluorescence techniques. The mean basal Mg(2+) uptake rate was 165 +/- 6 nM/s. ATP inhibited basal Mg(2+) uptake and hormone-stimulated Mg(2+) entry by 40%. Both P2X (P2X1-P2X5 subtypes) and P2Y2 receptor subtypes were identified in MDCT cells using differential RT-PCR. Activation of both receptor subtypes with selective agonists increased intracellular Ca(2+) concentration, P2X purinoceptors by ionotropic-gated channels, and P2Y receptors via G protein-mediated intracellular Ca(2+) release. The more relatively selective P2X agonists [beta,gamma-methylene ATP (beta,gamma-Me-ATP) and 2'- and -3'-O-(4-benzoyl-benzoyl)-ATP] inhibited arginine vasopressin (AVP)- and parathyroid hormone (PTH)-mediated Mg(2+) uptake whereas agonists more selective for P2Y purinoceptors (UTP, ADP, and 2-methylthio-ATP) were without effect. Removal of extracellular Ca(2+) diminished beta,gamma-Me-ATP-mediated increase in intracellular Ca(2+) and inhibition of AVP-stimulated Mg(2+) entry. We conclude from this information that ATP inhibited Mg(2+) uptake in MDCT cells through P2X purinoceptors expressed in this distal convoluted tubule cell line.