Luminal nucleotide stimulation is known to reduce Na+ transport in the distal nephron. Previous studies suggest that this mechanism may involve the thiazide-sensitive Na+-Cl- cotransporter (NCC), which plays an essential role in NaCl reabsorption in the cells lining the distal convoluted tubule (DCT). Here we show that stimulation of mouse DCT (mDCT) cells with ATP or UTP promoted Ca2+ transients and decreased the expression of NCC at both mRNA and protein levels. Specific siRNA-mediated silencing of P2Y2 receptors almost completely abolished ATP/UTP-induced Ca2+ transients and significantly reduced ATP/UTP-induced decrease of NCC expression. To test whether local variations in the intracellular Ca2+ concentration ([Ca2+]i) may control NCC transcription, we overexpressed the Ca2+-binding protein parvalbumin selectively in the cytosol or in the nucleus of mDCT cells. The decrease in NCC mRNA upon nucleotide stimulation was abolished in cells overexpressing cytosolic PV but not in cells overexpressing either a nuclear-targeted PV or a mutated PV unable to bind Ca2+. Using a firefly luciferase reporter gene strategy, we observed that the activity of NCC promoter region from -1 to -2,200 bp was not regulated by changes in [Ca2+]i. In contrast, high cytosolic calcium level induced instability of NCC mRNA. We conclude that in mDCT cells: (1) P2Y2 receptor is essential for the intracellular Ca2+ signaling induced by ATP/UTP stimulation; (2) P2Y2-mediated increase of cytoplasmic Ca2+ concentration down-regulates the expression of NCC; (3) the decrease of NCC expression occurs, at least in part, via destabilization of its mRNA.
Banderali U., Brochiero E., Lindenthal S., Raschi C., Bogliolo S., Ehrenfeld J., Control of apical membrane chloride permeability in the renal A6 cell line by nucleotides, 10.1111/j.1469-7793.1999.0737n.x
Belge H., Gailly P., Schwaller B., Loffing J., Debaix H., Riveira-Munoz E., Beauwens R., Devogelaer J.-P., Hoenderop J. G., Bindels R. J., Devuyst O., Renal expression of parvalbumin is critical for NaCl handling and response to diuretics, 10.1073/pnas.0702810104
Bidet M, De Renzis G, Martial S, Rubera I, Tauc M, Poujeol P (2000) Extracellular ATP increases [CA(2+)](i) in distal tubule cells. I. Evidence for a P2Y2 purinoceptor. Am J Physiol Renal Physiol 279(1):F92–F101
Burnstock G., Physiology and Pathophysiology of Purinergic Neurotransmission, 10.1152/physrev.00043.2006
Bustin S. A., Benes V., Garson J. A., Hellemans J., Huggett J., Kubista M., Mueller R., Nolan T., Pfaffl M. W., Shipley G. L., Vandesompele J., Wittwer C. T., The MIQE Guidelines: Minimum Information for Publication of Quantitative Real-Time PCR Experiments, 10.1373/clinchem.2008.112797
Dai Long-Jun, Kang Hyung Sub, Kerstan Dirk, Ritchie Gordon, Quamme Gary A., ATP inhibits Mg2+uptake in MDCT cells via P2X purinoceptors, 10.1152/ajprenal.0349.2000
Ellison David H., Brooks Virginia L., Renal Nerves, WNK4, Glucocorticoids, and Salt Transport, 10.1016/j.cmet.2011.05.007
Fuchs Sébastien, Philippe Josette, Corvol Pierre, Pinet Florence, Implication of Ref-1 in the repression of renin gene transcription by intracellular calcium : , 10.1097/00004872-200302000-00024
Gailly Philippe, Ca2+ entry in CHO cells, after Ca2+ stores depletion, is mediated by arachidonic acid, 10.1016/s0143-4160(98)90053-7
Gesek F A, Friedman P A, Mechanism of calcium transport stimulated by chlorothiazide in mouse distal convoluted tubule cells., 10.1172/jci115878
Gitelman HJ, Graham JB, Welt LG (1966) A new familial disorder characterized by hypokalemia and hypomagnesemia. Trans Assoc Am Physicians 79:221–235
Gordon J L, Extracellular ATP: effects, sources and fate, 10.1042/bj2330309
Huntzinger Eric, Izaurralde Elisa, Gene silencing by microRNAs: contributions of translational repression and mRNA decay, 10.1038/nrg2936
Ikura Mitsuhiko, Osawa Masanori, Ames James B., The role of calcium-binding proteins in the control of transcription: structure to function, 10.1002/bies.10105
Kaplan B., Davydov O., Knight H., Galon Y., Knight M. R., Fluhr R., Fromm H., Rapid Transcriptome Changes Induced by Cytosolic Ca2+ Transients Reveal ABRE-Related Sequences as Ca2+-Responsive cis Elements in Arabidopsis, 10.1105/tpc.106.042713
Kishore BK, Ginns SM, Krane CM, Nielsen S, Knepper MA (2000) Cellular localization of P2Y(2) purinoceptor in rat renal inner medulla and lung. Am J Physiol Renal Physiol 278(1):F43–F51
Klar J., Sigl M., Obermayer B., Schweda F., Kramer B. K., Kurtz A., Calcium Inhibits Renin Gene Expression by Transcriptional and Posttranscriptional Mechanisms, 10.1161/01.hyp.0000192025.86189.46
Lazarowski E. R., Mechanisms of Release of Nucleotides and Integration of Their Action as P2X- and P2Y-Receptor Activating Molecules, 10.1124/mol.64.4.785
Lazarowski Eduardo R, Harden T Kendall, Quantitation of extracellular UTP using a sensitive enzymatic assay, 10.1038/sj.bjp.0702654
Lazarowski ER, Rochelle LG, O'Neal WK, Ribeiro CM, Grubb BR, Zhang V, Harden TK, Boucher RC (2001) Cloning and functional characterization of two murine uridine nucleotide receptors reveal a potential target for correcting ion transport deficiency in cystic fibrosis gallbladder. J Pharmacol Exp Ther 297(1):43–49
Lehrmann H, Thomas J, Kim SJ, Jacobi C, Leipziger J (2002) Luminal P2Y2 receptor-mediated inhibition of Na+ absorption in isolated perfused mouse CCD. J Am Soc Nephrol 13(1):10–18
Livak Kenneth J., Schmittgen Thomas D., Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2−ΔΔCT Method, 10.1006/meth.2001.1262
Lu Ming, MacGregor Gordon G., Wang Wenhui, Giebisch Gerhard, Extracellular Atp Inhibits the Small-Conductance K Channel on the Apical Membrane of the Cortical Collecting Duct from Mouse Kidney, 10.1085/jgp.116.2.299
MacKenzie Steven, Vaitkevicius Henrikas, Lockette Warren, Sequencing and Characterization of the Human Thiazide-Sensitive Na-Cl Cotransporter (SLC12A3) Gene Promoter, 10.1006/bbrc.2001.4673
Misquitta Christine M., Chen Tao, Grover Ashok K., Control of protein expression through mRNA stability in calcium signalling, 10.1016/j.ceca.2006.04.004
Naveh-Many Tally, Nechama Morris, Regulation of parathyroid hormone mRNA stability by calcium, phosphate and uremia : , 10.1097/mnh.0b013e3281c55ede
Olinger E., Schwaller B., Loffing J., Gailly P., Devuyst O., Parvalbumin: calcium and magnesium buffering in the distal nephron, 10.1093/ndt/gfs457
Pochynyuk Oleh, Bugaj Vladislav, Rieg Timo, Insel Paul A., Mironova Elena, Vallon Volker, Stockand James D., Paracrine Regulation of the Epithelial Na+Channel in the Mammalian Collecting Duct by Purinergic P2Y2Receptor Tone, 10.1074/jbc.m807129200
Pochynyuk O., Rieg T., Bugaj V., Schroth J., Fridman A., Boss G. R., Insel P. A., Stockand J. D., Vallon V., Dietary Na+ inhibits the open probability of the epithelial sodium channel in the kidney by enhancing apical P2Y2-receptor tone, 10.1096/fj.09-151506
Pusl Thomas, Wu Julie J., Zimmerman Tracy L., Zhang Lei, Ehrlich Barbara E., Berchtold Martin W., Hoek Joannes B., Karpen Saul J., Nathanson Michael H., Bennett Anton M., Epidermal Growth Factor-mediated Activation of the ETS Domain Transcription Factor Elk-1 Requires Nuclear Calcium, 10.1074/jbc.m203002200
Rieg T., Bundey R. A., Chen Y., Deschenes G., Junger W., Insel P. A., Vallon V., Mice lacking P2Y2 receptors have salt-resistant hypertension and facilitated renal Na+ and water reabsorption, 10.1096/fj.07-8807com
Ross J (1995) mRNA stability in mammalian cells. Microbiol Rev 59(3):423–450
Rozen S, Skaletsky H (2000) Primer3 on the WWW for general users and for biologist programmers. Methods Mol Biol 132:365–386
Schaefer A, Magocsi M, Stocker U, Fandrich A, Marquardt H (1996) Ca2+/calmodulin-dependent and -independent down-regulation of c-myb mRNA levels in erythropoietin-responsive murine erythroleukemia cells. The role of calcineurin J Biol Chem 271(23):13484–13490
Schwiebert Erik M, Extracellular ATP and cAMP as Paracrine and Interorgan Regulators of Renal Function ATP Release Mechanisms, ATP Receptors and Purinergic Signalling along the Nephron, 10.1046/j.1440-1681.2001.03451.x
Sorensen Mads V., Grossmann Solveig, Roesinger Marian, Gresko Nikolay, Todkar Abhijeet P., Barmettler Gery, Ziegler Urs, Odermatt Alex, Loffing-Cueni Dominique, Loffing Johannes, Rapid dephosphorylation of the renal sodium chloride cotransporter in response to oral potassium intake in mice, 10.1038/ki.2013.14
Taboubi S., Milanini J., Delamarre E., Parat F., Garrouste F., Pommier G., Takasaki J., Hubaud J.-C., Kovacic H., Lehmann M., G (q/11)-coupled P2Y2 nucleotide receptor inhibits human keratinocyte spreading and migration, 10.1096/fj.06-7476com
Taniyama Yoshihiro, Sato Kazunori, Sugawara Akira, Uruno Akira, Ikeda Yukio, Kudo Masataka, Ito Sadayoshi, Takeuchi Kazuhisa, Renal Tubule-specific Transcription and Chromosomal Localization of Rat Thiazide-sensitive Na-Cl Cotransporter Gene, 10.1074/jbc.m101614200
Unwin RJ, Bailey MA, Burnstock G (2003) Purinergic signaling along the renal tubule: the current state of play. News Physiol Sci 18:237–241
Vallon V., Rieg T., Regulation of renal NaCl and water transport by the ATP/UTP/P2Y2 receptor system, 10.1152/ajprenal.00236.2011
Vargas-Poussou R., Dahan K., Kahila D., Venisse A., Riveira-Munoz E., Debaix H., Grisart B., Bridoux F., Unwin R., Moulin B., Haymann J.-P., Vantyghem M.-C., Rigothier C., Dussol B., Godin M., Nivet H., Dubourg L., Tack I., Gimenez-Roqueplo A.-P., Houillier P., Blanchard A., Devuyst O., Jeunemaitre X., Spectrum of Mutations in Gitelman Syndrome, 10.1681/asn.2010090907
Zambon AC, Hughes RJ, Meszaros JG, Wu JJ, Torres B, Brunton LL, Insel PA (2000) P2Y(2) receptor of MDCK cells: cloning, expression, and cell-specific signaling. Am J Physiol Renal Physiol 279(6):F1045–F1052
Bibliographic reference
Gailly, Philippe ; Szutkowska, Marta ; Olinger, Eric ; Debaix, Huguette ; Seghers, François ; et. al. P2Y2 receptor activation inhibits the expression of the sodium-chloride cotransporter NCC in distal convoluted tubule cells. In: Pflügers Archiv - European journal of physiology, Vol. 466, no. 11466(11):2035-47, p. 2035-2047 (2014)
Debaix, Huguette
[UCL]
Seghers, François
[UCL]
Janas, Sylvie
[UCL]
Devuyst, Olivier
[UCL]
