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Originally published In Press as doi:10.1074/jbc.M111880200 on January 8, 2002

J. Biol. Chem., Vol. 277, Issue 12, 10379-10386, March 22, 2002
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Long Term Regulation of Aquaporin-2 Expression in Vasopressin-responsive Renal Collecting Duct Principal Cells*

Udo HaslerDagger , David MordasiniDagger , Marcelle Bens§, Matthieu BianchiDagger , Françoise Cluzeaud§, Martine RousselotDagger , Alain Vandewalle§, Eric FérailleDagger ||, and Pierre-Yves MartinDagger ||

From the Dagger  Division of Nephrology, Fondation pour Recherches Médicales, 64 Avenue de la Roseraie, CH-1211, Genève 4, Switzerland and § INSERM U478, Faculté de Médecine Xavier Bichat, Institut Fédératif de Recherche 02, BP416, F-75870 Paris Cedex 18, France

Fine regulation of water reabsorption by the antidiuretic hormone [8-arginine]vasopressin (AVP) occurs in principal cells of the collecting duct and is largely dependent on regulation of the aquaporin-2 (AQP2) water channel. AVP-inducible long term AQP2 expression was investigated in immortalized mouse cortical collecting duct principal cells. Combined RNase protection assay, Western blot, and immunofluorescence analyses revealed that physiological concentrations of AVP added to the basal side, but not to the apical side, of cells grown on filters induced both AQP2 mRNA and apical protein expression. The stimulatory effect of AVP on AQP2 expression followed a V2 receptor-dependent pathway because [deamino-8-D-arginine]vasopressin (dDAVP), a specific V2 receptor agonist, produced the same effect as AVP, whereas the V2 antagonist SR121463B antagonized action of both AVP and dDAVP. Moreover, forskolin and cyclic 8-bromo-AMP fully reproduced the effects of AVP on AQP2 expression. Analysis of protein degradation pathways showed that inhibition of proteasomal activity prevented synthesis of AVP-inducible AQP2 mRNA and protein. Once synthesized, AQP2 protein was quickly degraded, a process that involves both the proteasomal and lysosomal pathways. This is the first study that delineates induction and degradation mechanisms of AQP2 endogenously expressed by a renal collecting duct principal cell line.


* This work was supported in part by the Swiss National Science Foundation Grant 31-56504.99 (to P.-Y. M.), by a grant from the Fondation Carlos et Elsie de Reuter (to P.-Y. M. and E. F.), by a grant from the Department of Medicine of Geneva University Hospital (to P.-Y. M.), and by INSERM.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 U478, Faculté de Médecine Xavier Bichat, BP416, F-75870 Paris Cedex 18, France. Tel.: 33-1-44-85-63-21; Fax: 33-1-42-29-16-44; E-mail: vandewal@bichat.inserm.fr.

|| Both authors contributed equally to this work.


Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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