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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
Institut de Pharmacologie et de Toxicologie de
l'Université, Lausanne, Switzerland
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 
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 
-ENaC mRNA (but not in those for ß- or 
-ENaC)
and three- to 10-fold increases in -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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 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. [Abstract] [Full Text] [PDF]
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