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. Extracellular nucleotides regulate renal ion transport. With the use of  in vitro  perfusion and [Ca 2+ ] i  imaging, this study investigated whether mouse and rabbit cortical collecting ducts (CCD) respond to luminal nucleotides. In mouse CCD, luminal ATP (EC 50 : 10 μM) and UTP (EC 50 : 9.7 μM) increased [Ca 2+ ] i  with an initial peak and a plateau. To make certain that basolateral P2 receptors were not activated by luminal nucleotides via leak diffusion, luminal trypsin (1 μM), a known agonist for basolateral proteinase-activated receptors, was perfused. Mouse CCD that were responsive to luminal ATP were nonresponsive to luminal trypsin but always showed [Ca 2+ ] i  elevations by basolateral trypsin (10 or 100 nM). Luminal α,β- and β,γ-methylene ATP, 2-methyl-S-ATP, ADP, UDP, and 2′,3′-O-4-benzoylbenzoyl ATP had no effect (100 μM,  n  = 9). Without external Ca 2+ , luminal ATP still stimulated a [Ca 2+ ] i  increase. Mouse CCD also responded to basolateral ATP (EC 50 : 23 μM) and UTP (EC 50 : 23 μM) with smaller [Ca 2+ ] i  elevations. Confocal microscopy of perfused CCD showed that luminal ATP (100 μM) rapidly increased [Ca 2+ ] i  in nearly all cells ( n  = 6) and the same cells that responded to luminal ATP responded to basolateral ATP (100 μM). In contrast, rabbit CCD did not respond to luminal ATP/UTP ( n  = 8) despite ATP's known effect from the basolateral side (EC 50 : 34 μM). These data indicate the expression of luminal P2Y receptors (probably P2Y 2 ) in principal cells of mouse CCD but not in rabbit CCD.

The Luminal P2Y Receptor in the Isolated Perfused Mouse Cortical Collecting Duct