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A more recent version of this article appeared on October 20, 2000
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M006591200v1
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Papers In Press, published online ahead of print August 14, 2000
J. Biol. Chem, 10.1074/jbc.M006591200
Submitted on July 24, 2000
Accepted on August 7, 2000

Regulation by glucocorticoids of expression and activity of rBSC1, the Na+-K+(NH4+)-2Cl- cotransporter of medullary thick ascending limb

Amel Attmane-Elakeb, Valérie Sibella, Catherine Vernimmen, Xavier Belenfant, Steven C. Hebert, and Maurice M. Bichara

INSERM U.426, Faculté de Médecine Xavier Bichat, Paris 75870 Cedex 18

Corresponding Author: bichara{at}bichat.inserm.fr

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, adrenalectomized (ADX), and ADX rats infused with dexamethasone (ADX+Dexa) 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 by quantitative RT-PCR. The abundance of rBSC1 protein and mRNA increased in ADX+Dexa 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 H2O) containing 0.7 nM arginine vasopressin (AVP), 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.


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