Generation and phenotype of mice harboring a nonsense mutation in the V2 vasopressin receptor gene
June Yun, … , Chu-xia Deng, Jürgen Wess
June Yun, … , Chu-xia Deng, Jürgen Wess
Published December 1, 2000
Citation Information: J Clin Invest. 2000;106(11):1361-1371. https://doi.org/10.1172/JCI9154.
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Article

Generation and phenotype of mice harboring a nonsense mutation in the V2 vasopressin receptor gene

Abstract

The V2 vasopressin receptor (V2R) plays a key role in the maintenance of a normal body water balance. To generate an in vivo model that allows the physiological and molecular analysis of the role of V2Rs in kidney function, we have created mouse lines that lack functional V2Rs by using targeted mutagenesis in mouse embryonic stem cells. Specifically, we introduced a nonsense mutation known to cause X-linked nephrogenic diabetes insipidus (XNDI) in humans (Glu242stop) into the mouse genome. V2R-deficient hemizygous male pups showed a decrease in basal urine osmolalities and were unable to concentrate their urine. These pups also exhibited an enlargement of renal pelvic space, failed to thrive, and died within the first week after birth due to hypernatremic dehydration. Interestingly, female mice heterozygous for the V2R mutation showed normal growth but displayed an XNDI-like phenotype, characterized by reduced urine concentrating ability of the kidney, polyuria, and polydipsia. Western blot analysis and immunoelectron microscopic studies showed that the loss of functional V2Rs had no significant effect on the basal expression levels of aquaporin-2 and the bumetanide-sensitive Na-K-2Cl cotransporter (BSC-1). The V2R mutant mice described here should serve as highly useful tools for the development of novel therapeutic strategies for the treatment of XNDI.

Authors

June Yun, Torsten Schöneberg, Jie Liu, Angela Schulz, Carolyn A. Ecelbarger, Dominique Promeneur, Soren Nielsen, Hui Sheng, Alexander Grinberg, Chu-xia Deng, Jürgen Wess

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Figure 1

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Targeted mutagenesis of the mouse V2R gene. (a) General strategy. The st...
Targeted mutagenesis of the mouse V2R gene. (a) General strategy. The structures of the V2R targeting construct, the wild-type V2R locus, and the targeted mutant V2R allele are shown. V2R coding sequences are depicted as open boxes. The neo cassette is represented by a hatched box, and the loxP sites are shown as arrows at either end of the neo cassette. The BamHI (b) sites used for Southern blot analysis and the sizes of the restriction fragments detected by using the 3′ probe are indicated. The Glu242stop nonsense mutation (GAA→TAG) that was introduced into exon 2 is represented by an asterisk. This nonsense mutation was generated in a fashion to introduce a novel NheI site at the locus of Glu242stop mutation. The targeting construct contained approximately 6 kb (5′ arm) and 1.4 kb (3′ arm) of genomic sequence, respectively. E, EcoRI; H, HpaI. (b) Genotyping of F2 mouse offspring via Southern blot analysis of BamHI-digested mouse tail DNA by using the 3′ probe. The 3.7- and 2.0-kb bands represent the wild-type and mutant V2R alleles, respectively. (c) Verification of the presence of the Glu242stop mutation in F2 offspring using PCR analysis of genomic tail DNA, followed by NheI digestion of the PCR product (for details, see Methods). The 350-bp band represents the uncut PCR product, indicative of the wild-type V2R allele; the 290-bp fragment is produced by NheI cleavage and is indicative of the mutant V2R allele. The smaller NheI cleavage product, 60 bp in size, is not shown.