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Regulation of the vasopressin V2 receptor by vasopressin in polarized renal collecting duct cells

Robben, J. H., Knoers, N. V. A. M. & Deen, P. M. T., Dec-2004, In : Molecular Biology of the Cell. 15, 12, p. 5693-9 7 p.

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  • Regulation of the Vasopressin V2 Receptor by Vasopressin in Polarized Renal Collecting Duct Cells

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DOI

Binding of arginine-vasopressin (AVP) to its V2 receptor (V2R) in the basolateral membrane of principal cells induces Aquaporin-2-mediated water reabsorption in the kidney. To study the regulation of the V2R by dDAVP in a proper model, a polarized renal cell line stably-expressing V2R-GFP was generated. Labeled AVP-binding studies revealed an equal basolateral vs. apical membrane distribution for V2R-GFP and endogenous V2R. In these cells, GFP-V2R was expressed in its mature form and localized for 75% in the basolateral membrane and for 25% to late endosomes/lysosomes. dDAVP caused a dose- and time-dependent internalization of V2R-GFP, which was completed within 1 h with 100 nM dDAVP, was prevented by coincubation with a V2R antagonist, and which reduced its half-life from 11.5 to 2.8 h. Semiquantification of the V2R-GFP colocalization with E-cadherin (basolateral membrane), early endosomal antigen-1 (EEA-1) and lysosome-associated membrane protein-2 (LAMP-2) in time revealed that most dDAVP-bound V2R was internalized via early endosomes to late endosomes/lysosomes, where it was degraded. The dDAVP-internalized V2R did not recycle to the basolateral membrane. In conclusion, we established the itinerary of the V2R in a polarized cell model that likely resembles the in vivo V2R localization and regulation by AVP to a great extent.

Original languageEnglish
Pages (from-to)5693-9
Number of pages7
JournalMolecular Biology of the Cell
Volume15
Issue number12
Publication statusPublished - Dec-2004
Externally publishedYes

    Keywords

  • Animals, Cell Line, Cell Polarity, Dogs, Glycosylation, Kidney Tubules, Collecting/cytology, Lysosomes/drug effects, Organelles/metabolism, Protein Processing, Post-Translational/drug effects, Receptors, Vasopressin/genetics, Time Factors, Vasopressins/pharmacology

ID: 92706089