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J. Biol. Chem., Vol. 275, Issue 43, 33548-33553, October 27, 2000
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From the To assess whether glucocorticoids regulate rBSC1,
the apical
Na+-K+(NH4+)-2Cl
Regulation by Glucocorticoids of Expression and Activity of
rBSC1, the
Na+-K+(NH4+)-2Cl
Cotransporter of Medullary Thick Ascending Limb*
§,,,
**, and
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
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.
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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