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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
Long Term Regulation of Aquaporin-2 Expression in
Vasopressin-responsive Renal Collecting Duct Principal Cells*
Udo
Hasler ,
David
Mordasini ,
Marcelle
Bens§,
Matthieu
Bianchi ,
Françoise
Cluzeaud§,
Martine
Rousselot ,
Alain
Vandewalle§¶,
Eric
Féraille , and
Pierre-Yves
Martin
From the 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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Copyright © 2002 by the American Society for Biochemistry and Molecular Biology.
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