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J Am Soc Nephrol 10:923-934, 1999
© 1999 American Society of Nephrology

REGULAR ARTICLES

Corticosteroid-Dependent Sodium Transport in a Novel Immortalized Mouse Collecting Duct Principal Cell Line

MARCELLE BENS*, VÉRONIQUE VALLET{dagger}, FRANÇOISE CLUZEAUD*, LAURENT PASCUAL-LETALLEC*, AXEL KAHN{ddagger}, MARIE E. RAFESTIN-OBLIN*, BERNARD C. ROSSIER{dagger} and ALAIN VANDEWALLE*

* Institut National de la Santé et de la Recherche Médicale (INSERM) Unité 478, Institut Fédératif de Recherche 02, Faculté de Médecine Xavier Bichat, Paris, Cedex, France
{dagger} Institut de Pharmacologie et de Toxicologie de l'Université, Lausanne, Switzerland
{ddagger} INSERM Unité 129, Faculté Cochin, Paris, France

Correspondence to Dr. Alain Vandewalle, Institut National de la Sante et de la Recherche Medicale U478, Faculté de Médecine Xavier Bichat, B.P. 416, 75870 Paris Cedex 18, France. Phone: 33 1 44856326; Fax: 33 1 42291644; E-mail: vandewal{at}bichat.inserm.fr

Abstract. The final control of sodium balance takes place in the cortical collecting duct (CCD) of the nephron, where corticosteroid hormones regulate sodium reabsorption by acting through mineralocorticoid (MR) and/or glucocorticoid (GR) receptors. A clone of principal CCD cells (mpkCCDc14) has been established that is derived from a transgenic mouse (SV40 large T antigen under the control of the SV40 enhancer/L-type pyruvate kinase promoter). Cells grown on filters form polarized monolayers with high electrical transepithelial resistance (RT approximately 4700 {Omega} x cm2) and potential difference (PD approximately -50 mV) and have an amiloride-sensitive electrogenic sodium transport, as assessed by the short-circuit current method (Isc approximately 11 µA/cm2). Reverse transcription-PCR experiments using rat MR primers, [3H]aldosterone, and [3H]dexamethasone binding and competition studies indicated that the mpkCCDc14 cells exhibit specific MR and GR. Aldosterone increased Isc in a dose- (10-10 to 10-6 M) and time-dependent (2 to 72 h) manner, whereas corticosterone only transiently increased Isc (2 to 6 h). Consistent with the expression of 11ß-hydroxysteroid dehydrogenase type 2, which metabolizes glucocorticoids to inactive 11-dehydroderivates, carbenoxolone potentiated the corticosterone-stimulated Isc. Aldosterone (5 x 10-7 M)-induced Isc (fourfold) was associated with a three- to fivefold increase in {alpha}-ENaC mRNA (but not in those for ß- or {gamma}-ENaC) and three- to 10-fold increases in {alpha}-ENaC protein synthesis. In conclusion, this new immortalized mammalian CCD clonal cell line has retained a high level of epithelial differentiation and sodium transport stimulated by aldosterone and therefore represents a useful mammalian cell system for identifying the genes controlled by aldosterone.




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A. Ouvrard-Pascaud, S. Puttini, Y. Sainte-Marie, R. Athman, V. Fontaine, F. Cluzeaud, N. Farman, M.-E. Rafestin-Oblin, M. Blot-Chabaud, and F. Jaisser
Conditional gene expression in renal collecting duct epithelial cells: use of the inducible Cre-lox system
Am J Physiol Renal Physiol, January 1, 2004; 286(1): F180 - F187.
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 Am. J. Physiol. Renal Physiol.Home page
M. Auberson, N. Hoffmann-Pochon, A. Vandewalle, S. Kellenberger, and L. Schild
Epithelial Na+ channel mutants causing Liddle's syndrome retain ability to respond to aldosterone and vasopressin
Am J Physiol Renal Physiol, September 1, 2003; 285(3): F459 - F471.
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 Mol. Biol. CellHome page
M. Vinciguerra, G. Deschenes, U. Hasler, D. Mordasini, M. Rousselot, A. Doucet, A. Vandewalle, P.-Y. Martin, and E. Feraille
Intracellular Na+ Controls Cell Surface Expression of Na,K-ATPase via a cAMP-independent PKA Pathway in Mammalian Kidney Collecting Duct Cells
Mol. Biol. Cell, July 1, 2003; 14(7): 2677 - 2688.
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 J. Biol. Chem.Home page
U. Hasler, D. Mordasini, M. Bianchi, A. Vandewalle, E. Feraille, and P.-Y. Martin
Dual Influence of Aldosterone on AQP2 Expression in Cultured Renal Collecting Duct Principal Cells
J. Biol. Chem., June 6, 2003; 278(24): 21639 - 21648.
[Abstract] [Full Text] [PDF]

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 Mol. Pharmacol.Home page
M.-E. Rafestin-Oblin, J. Fagart, A. Souque, C. Seguin, M. Bens, and A. Vandewalle
11beta -Hydroxyprogesterone Acts as a Mineralocorticoid Agonist in Stimulating Na+ Absorption in Mammalian Principal Cortical Collecting Duct Cells
Mol. Pharmacol., December 1, 2002; 62(6): 1306 - 1313.
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 JGPHome page
G. Vuagniaux, V. Vallet, N. F. Jaeger, E. Hummler, and B. C. Rossier
Synergistic Activation of ENaC by Three Membrane-bound Channel-activating Serine Proteases (mCAP1, mCAP2, a