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

MARCELLE BENS, VÉRONIQUE VALLET, FRANÇOISE CLUZEAUD, LAURENT PASCUAL-LETALLEC, AXEL KAHN, MARIE E. RAFESTIN-OBLIN, BERNARD C. ROSSIER and ALAIN VANDEWALLE
JASN May 1999, 10 (5) 923-934;
MARCELLE BENS
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VÉRONIQUE VALLET
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FRANÇOISE CLUZEAUD
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LAURENT PASCUAL-LETALLEC
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AXEL KAHN
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MARIE E. RAFESTIN-OBLIN
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BERNARD C. ROSSIER
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ALAIN VANDEWALLE
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Abstract

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 Ω × 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 11β-hydroxysteroid dehydrogenase type 2, which metabolizes glucocorticoids to inactive 11-dehydroderivates, carbenoxolone potentiated the corticosterone-stimulated Isc. Aldosterone (5 × 10-7 M)-induced Isc (fourfold) was associated with a three- to fivefold increase in α-ENaC mRNA (but not in those for β- or γ-ENaC) and three- to 10-fold increases in α-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.

  • © 1999 American Society of Nephrology
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Journal of the American Society of Nephrology: 10 (5)
Journal of the American Society of Nephrology
Vol. 10, Issue 5
1 May 1999
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Corticosteroid-Dependent Sodium Transport in a Novel Immortalized Mouse Collecting Duct Principal Cell Line
MARCELLE BENS, VÉRONIQUE VALLET, FRANÇOISE CLUZEAUD, LAURENT PASCUAL-LETALLEC, AXEL KAHN, MARIE E. RAFESTIN-OBLIN, BERNARD C. ROSSIER, ALAIN VANDEWALLE
JASN May 1999, 10 (5) 923-934;

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Corticosteroid-Dependent Sodium Transport in a Novel Immortalized Mouse Collecting Duct Principal Cell Line
MARCELLE BENS, VÉRONIQUE VALLET, FRANÇOISE CLUZEAUD, LAURENT PASCUAL-LETALLEC, AXEL KAHN, MARIE E. RAFESTIN-OBLIN, BERNARD C. ROSSIER, ALAIN VANDEWALLE
JASN May 1999, 10 (5) 923-934;
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More in this TOC Section

  • Acute Regulation of the Epithelial Na+ Channel by Phosphatidylinositide 3-OH Kinase Signaling in Native Collecting Duct Principal Cells
  • Role for TGF-β in Cyclosporine-Induced Modulation of Renal Epithelial Barrier Function
  • Increased Renal Responsiveness to Vasopressin and Enhanced V2 Receptor Signaling in RGS2−/− Mice
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