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Am J Physiol Renal Physiol 283: F437-F446, 2002. First published May 14, 2002; doi:10.1152/ajprenal.00316.2001
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Vol. 283, Issue 3, F437-F446, September 2002

Effects of luminal flow and nucleotides on [Ca2+]i in rabbit cortical collecting duct

Craig B. Woda1, Maurilo Leite Jr.2, Rajeev Rohatgi3, and Lisa M. Satlin1,3

Departments of 1 Pediatrics and 3 Medicine, Mount Sinai School of Medicine, New York, New York 10029-6574; and 2 Department of Medicine, Universidade Federal do Rio de Janeiro, Hospital Universitario Clementino Fraga Filho, 21949-900 Rio de Janeiro, Brazil

Nucleotide binding to purinergic P2 receptors contributes to the regulation of a variety of physiological functions in renal epithelial cells. Whereas P2 receptors have been functionally identified at the basolateral membrane of the cortical collecting duct (CCD), a final regulatory site of urinary Na+, K+, and acid-base excretion, controversy exists as to whether apical purinoceptors exist in this segment. Nor has the distribution of receptor subtypes present on the unique cell populations that constitute Ca2+ the CCD been established. To examine this, we measured nucleotide-induced changes in intracellular Ca2+ concentration ([Ca2+]i) in fura 2-loaded rabbit CCDs microperfused in vitro. Resting [Ca2+]i did not differ between principal and intercalated cells, averaging ~120 nM. An acute increase in tubular fluid flow rate, associated with a 20% increase in tubular diameter, led to increases in [Ca2+]i in both cell types. Luminal perfusion of 100 µM UTP or ATP-gamma -S, in the absence of change in flow rate, caused a rapid and transient approximately fourfold increase in [Ca2+]i in both cell types (P < 0.05). Luminal suramin, a nonspecific P2 receptor antagonist, blocked the nucleotide- but not flow-induced [Ca2+]i transients. Luminal perfusion with a P2X (alpha ,beta -methylene-ATP), P2X7 (benzoyl-benzoyl-ATP), P2Y1 (2-methylthio-ATP), or P2Y4/P2Y6 (UDP) receptor agonist had no effect on [Ca2+]i. The nucleotide-induced [Ca2+]i transients were inhibited by the inositol-1,4,5-triphosphate receptor blocker 2-aminoethoxydiphenyl borate, thapsigargin, which depletes internal Ca2+ stores, luminal perfusion with a Ca2+-free perfusate, or the L-type Ca2+ channel blocker nifedipine. These results suggest that luminal nucleotides activate apical P2Y2 receptors in the CCD via pathways that require both internal Ca2+ mobilization and extracellular Ca2+ entry. The flow-induced rise in [Ca2+]i is apparently not mediated by apical P2 purinergic receptor signaling.

microperfusion; fura 2; purinergic receptor; principal cell; intercalated cell; intracellular calcium concentration


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