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Originally published In Press as doi:10.1074/jbc.M006591200 on August 14, 2000

J. Biol. Chem., Vol. 275, Issue 43, 33548-33553, October 27, 2000
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Regulation by Glucocorticoids of Expression and Activity of rBSC1, the Na+-K+(NH4+)-2Clminus Cotransporter of Medullary Thick Ascending Limb*

Amel Attmane-ElakebDagger §, Valérie SibellaDagger , Catherine VernimmenDagger , Xavier Belenfant, Steven C. Hebert||**, and Maurice BicharaDagger Dagger Dagger

From the Dagger  INSERM U.426, Institut Fédératif Régional Xavier Bichat, Faculté de Médecine Xavier Bichat, Université Paris 7, 75870 Paris Cédex 18, France, the  Service de Néphrologie, Hôpital André Grégoire, 93100 Montreuil, France, and the || Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, Connecticut 06520-8026

To assess whether glucocorticoids regulate rBSC1, the apical Na+-K+(NH4+)-2Cl- cotransporter of kidney medullary thick ascending limb (MTAL), studies were performed in normal rats, adrenalectomized (ADX) rats, and ADX rats infused with dexamethasone for 6 days. The effects of dexamethasone on rBSC1 were also studied in vitro using isolated rat MTAL segments. Cotransport activity was estimated by intracellular pH measurements; rBSC1 protein was quantified in MTAL crude membranes by immunoblotting analysis, and mRNA was quantified by quantitative reverse transcription-polymerase chain reaction. The abundance of rBSC1 protein and mRNA increased in ADX rats infused with dexamethasone compared with ADX rats (p < 0.04). In addition, application of dexamethasone for 1-3 h to MTALs caused rBSC1 protein and mRNA abundance and cotransport activity to significantly increase in a hyperosmotic medium (450 mosmol/kg of H2O) containing 0.7 nM arginine vasopressin, which is an in vitro experimental condition that resembles the in vivo MTAL environment. Results obtained in various media and with 8-bromo-cAMP indicated that stimulation of rBSC1 expression by glucocorticoids required interactions between glucocorticoid receptor- and cAMP-dependent factors. Up to 100 nM d-aldosterone had no effect on cotransport activity in vitro. Thus glucocorticoids directly stimulate MTAL rBSC1 expression and activity, which contributes to glucocorticoid-dependent effects on the renal regulation of acid-base balance and urinary concentrating ability.

* This work was supported in part by grants from the Institut National de la Recherche Médicale, the Universités Paris 6 and Paris 7, the Fondation pour la Recherche Médicale Française, and the Fondation de France.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ Supported by a grant from La Ligue Nationale Contre Le Cancer.

** Supported by National Institutes of Health Grant DK36803.

Dagger Dagger To whom correspondence should be addressed: INSERM U.426. Faculté de Médecine Xavier Bichat, 16 rue Henri Huchard, 75870 Paris cédex 18, France. Tel.: 33 1 44 85 62 81; Fax: 33 1 42 28 15 64; E-mail: bichara@bichat.inserm.fr.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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