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Published ahead of print on September 7, 2006
J Am Soc Nephrol 17: 2680-2686, 2006
© 2006 American Society of Nephrology
doi: 10.1681/ASN.2006030246
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Cell and Transport Physiology

Vasopressin Increases Plasma Membrane Accumulation of Urea Transporter UT-A1 in Rat Inner Medullary Collecting Ducts

Janet D. Klein*, Otto Fröhlich{dagger}, Mitsi A. Blount*, Christopher F. Martin*, Tekla D. Smith* and Jeff M. Sands*,{dagger}

* Renal Division, Department of Medicine; and {dagger} Department of Physiology, Emory University School of Medicine, Atlanta, Georgia

Address correspondence to: Dr. Janet D. Klein, Emory University School of Medicine, Renal Division, 1639 Pierce Drive NE, WMB Room 3319B, Atlanta, GA 30322. Phone: 404-727-9933; Fax: 404-727-3425; E-mail: janet.klein{at}emory.edu

Received for publication March 20, 2006. Accepted for publication July 25, 2006.

Urea transport, mediated by the urea transporter A1 (UT-A1) and/or UT-A3, is important for the production of concentrated urine. Vasopressin rapidly increases urea transport in rat terminal inner medullary collecting ducts (IMCD). A previous study showed that one mechanism for rapid regulation of urea transport is a vasopressin-induced increase in UT-A1 phosphorylation. This study tests whether vasopressin or directly activating adenylyl cyclase with forskolin also increases UT-A1 accumulation in the plasma membrane of rat IMCD. Inner medullas were harvested from rats 45 min after injection with vasopressin or vehicle. UT-A1 abundance in the plasma membrane was significantly increased in the membrane fraction after differential centrifugation and in the biotinylated protein population. Vasopressin and forskolin each increased the amount of biotinylated UT-A1 in rat IMCD suspensions that were treated ex vivo. The observed changes in the plasma membrane are specific, as the amount of biotinylated UT-A1 but not the calcium-sensing receptor was increased by forskolin. Next, whether forskolin or the V2-selective agonist dDAVP would increase apical membrane expression of UT-A1 in MDCK cells that were stably transfected with UT-A1 (UT-A1-MDCK cells) was tested. Forskolin and dDAVP significantly increased UT-A1 abundance in the apical membrane in UT-A1-MDCK cells. It is concluded that vasopressin and forskolin increase UT-A1 accumulation in the plasma membrane in rat IMCD and in the apical plasma membrane of UT-A1-MDCK cells. These findings suggest that vasopressin regulates urea transport by increasing UT-A1 accumulation in the plasma membrane and/or UT-A1 phosphorylation.




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