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Am J Physiol Cell Physiol 285: C1445-C1453, 2003. First published August 13, 2003; doi:10.1152/ajpcell.00386.2002
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RECEPTORS AND SIGNAL TRANSDUCTION

Purinergic-induced signaling in C11-MDCK cells inhibits the secretory Na-K-Cl cotransporter

Tatyana A. Brindikova,3 Nathalie Bourcier,2 Brian Torres,4 Dimitri Pchejetski,2,3 Michel Gekle,5 Georgy V. Maximov,3 Valérie Montminy,1 Paul A. Insel,4 Sergei N. Orlov,2,3,* and Paul Isenring1,*

1Nephrology Group, Research Center L'Hôtel-Dieu de Québec, Department of Medicine, Faculty of Medicine, Laval University, Québec G1R 2J6; 2Research Centre l'Hôtel-Dieu de Montréal, Centre Hospitalier de l'Université de Montréal, Montréal, Quebec, Canada H2W 1T8; 3Faculty of Biology, MV Lomonosov Moscow State University, Moscow, Russia; 4Department of Pharmacology, University of California, San Diego, La Jolla, California 92093; and 5Department of Physiology, University of Wurzburg, D97070 Wurzburg, Germany

Submitted 25 August 2002 ; accepted in final form 4 August 2003

Purinergic inhibition of Na-K-Cl cotransport has been noted in various renal epithelial cells derived from the collecting tubule, including Madin-Darby canine kidney (MDCK) cells. In recent studies, we have observed purinergic inhibition of Na-K-Cl cotransport in C11-MDCK subclones ({alpha}-intercalated-like cells). Interestingly, Na-K-Cl cotransport activity was also detected in C7-MDCK subclones (principal-like cells) but was not affected by ATP. In this investigation, we have transfected the human Na-K-Cl cotransporter (huNKCC1) in both C11 and C7 cells to determine whether these differences in NKCC regulation by ATP were due to cell-specific purinoceptor signaling pathways or to cell-specific isoforms/splice variants of the transporter. In both cell lines, we found that endogenous as well as huNKCC1-derived cotransport activity was restricted to the basolateral side. In addition, we were able to show that extracellular application of 100 µM ATP or 100 µM UTP abolished NKCC activity in both mock- and huNKCC1-transfected C11 cells but not in mock- and huNKCC1-transfected C7 cells; in C11 cells, intriguingly, this inhibition was not affected by inhibitors of RNA and protein synthesis and occurred even though expression levels of UTP-sensitive P2Y2-, P2Y4-, and P2Y6-purinoceptors were not different from those observed in C7 cells. These results suggest that C11 cells express an undetermined type of UTP-sensitive P2-purinoceptors or a unique P2Y-purinoceptor-triggered signaling cascade that leads to inhibition of NKCC1.

P2-purinoceptors; Na-K-Cl cotransporter; C7- and C11-Madin-Darby canine kidney cells


Address for reprint requests and other correspondence: P. Isenring, L'Hôtel-Dieu de Québec Research Center, 10 rue McMahon (Rm. 3852), QC, Canada G1R 2J6 (E-mail: paul.isenring{at}crhdq.ulaval.ca).




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