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Am J Physiol Renal Physiol 279: F102-F111, 2000;
0363-6127/00 $5.00
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Vol. 279, Issue 1, F102-F111, July 2000

Extracellular ATP increases [Ca2+]i in distal tubule cells. II. Activation of a Ca2+-dependent Clminus conductance

Isabelle Rubera, Michel Tauc, Michel Bidet, Catherine Verheecke-Mauze, Guy De Renzis, Chantal Poujeol, Béatrice Cuiller, and Philippe Poujeol

Unité Mixte de Recherche 6548, Centre National de la Recherche Scientifique, Université de Nice-Sophia Antipolis, 06108 Nice Cedex 2, France

We characterized Cl- conductance activated by extracellular ATP in an immortalized cell line derived from rabbit distal bright convoluted tubule (DC1). 125I- efflux experiments showed that ATP increased 125I- loss with an EC50 = 3 µM. Diphenylamine-2-carboxylate (10-3 M) and NPPB (10-4 M) abolished the 125I- efflux. Preincubation with 10 µM 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester or 10-7 M thapsigargin inhibited the effect of ATP. Ionomycin (2 µM) increased 125I- efflux with a time course similar to that of extracellular ATP, suggesting that the response is dependent on the intracellular Ca2+ concentration ([Ca2+]i). The ATP agonist potency order was ATP >=  UTP > ATPgamma S. Suramin (500 µM) inhibited the ATP-induced 125I- efflux, consistent with P2 purinoceptors. 125I- effluxes from cells grown on permeable filters suggest that ATP induced an apical efflux that was mediated via apical P2 receptors. Whole cell experiments showed that ATP (100 µM) activated outwardly rectifying Cl- currents in the presence of 8-cyclopentyl-1,3-dipropylxanthine, excluding the involvement of P1 receptors. Ionomycin activated Cl- currents similar to those developed with ATP. These results demonstrate the presence of a purinergic regulatory mechanism involving ATP, apical P2Y2 receptors, and Ca2+ mobilization for apical Cl- conductance in a distal tubule cell line.

kidney; intracellular calcium


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