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Articles in PresS, published online ahead of print May 10, 2002
Am J Physiol Renal Physiol, 10.1152/ajprenal.00316.2001
Submitted on October 12, 2001
Accepted on March 12, 2002
1 Department of Pediatrics, Mount Sinai School of Medicine, New York, New York, USA
2 Department of Medicine, Universidade Federal do Rio de Janeiro, Rio De Janeiro, Brazil
3 Department of Medicine, Mount Sinai School of Medicine, New York, New York, USA
4 Department of Pediatrics, Mount Sinai School of Medicine, New York, New York, USA; Medicine, Mount Sinai School of Medicine, New York, New York, USA
* To whom correspondence should be addressed. E-mail: lisa.satlin{at}mssm.edu.
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 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 comprise the CCD been established. To examine this, we measured nucleotide-induced changes in intracellular calcium 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-
-S, in the absence of change in flow rate, caused a rapid and transient ~4-fold 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 (
,ß-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 IP3 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.
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