PMC:1189073 / 3078-6695 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/1189073","sourcedb":"PMC","sourceid":"1189073","source_url":"http://www.ncbi.nlm.nih.gov/pmc/1189073","text":"Introduction\nNephrogenic diabetes insipidus (NDI) is a disease characterized by excessive urination and thirst, despite normal production of the antidiuretic hormone arginine vasopressin (AVP) [1]. The inherited forms are either X-linked as a consequence of mutation of the Avpr2 gene [2], or autosomal due to mutation of the Aqp2 gene [3]. Aquaporin-2 (AQP2) is a pore-forming protein belonging to a family of water channels [4], and it is expressed in collecting-duct principal cells in the kidney [5]. Generally these proteins permit the passage of water through the plasma membrane (PM) of cells, several of which carry out this role specifically in the process of water reabsorption from urine in the kidney. It has been established that aquaporins, although functional as a monomer, tetramerize before their insertion into the plasma membrane [4,6]. Furthermore these proteins can also be differentially targeted to distinct regions of the PM; for example, AQP2 is routed to the apical membrane of cells surrounding the collecting duct, whereas other aquaporins (AQP3 or 4) are inserted into the basolateral face. Unlike all other family members, AQP2 is not constitutively inserted into the plasma membrane. Under basal conditions, the protein resides in subapical intracellular vesicles; however, under conditions requiring water retention AQP2 translocates to the apical membrane, permitting water reabsorption [7,8]. For this process to occur, AVP binds its receptor, AVPR2, on the basolateral face of the collecting duct cells, leading to a rise in intracellular cAMP, ultimately resulting in phosphorylation of AQP2 at serine 256 by cAMP-dependent protein kinase [9] and its redistribution to the plasma membrane.\nThe importance of AQP2 redistribution has been highlighted by functional characterization of Aqp2 mutations resulting in severe NDI in humans [3,10]. Recessive Aqp2 mutations are generally thought to produce an abnormally localized and, in most instances, misfolded water pore that responds abnormally to an increase in cAMP [6,11]. Furthermore, dominant mutations have been described and found to misroute both the mutant and the wild-type protein to the basolateral membrane [6,12].\nSeveral mouse models of diabetes insipidus have been generated [13–17]. In an attempt to recapitulate human NDI, mice have been generated with mutations in Aqp2 and Avpr2 [15,18]. Yang and colleagues created a mouse with a T126M knock-in mutation in the Aqp2 gene. Unexpectedly, homozygous mutant mice died within 6 d after birth. Interestingly, AVPR2-deficient male pups also die within the first week after birth. Together these models suggest that the mouse may be a highly sensitive organism with regard to water homeostasis, and is unable to survive with polyuria.\nIn a forward genetic screen, a mouse with an Aqp2 mutation was identified. The purpose of this study was to characterize this murine model of recessive nephrogenic DI. We now report a novel F204V mutation in the Aqp2 gene. This allele of Aqp2 was found to cause the first mouse model of NDI to survive past the first week of life. Molecular analyses concluded that mutant AQP2 adopts a different subcellular localization in renal collecting-duct cells, and was resistant to translocation induced by desmopressin, an agonist of AVP. In vitro studies using the Madin-Darby canine kidney (MDCK) cell line demonstrated an endoplasmic reticulum pattern for the mutant protein, and apparent resistance to translocation. These data conclusively prove that autosomal recessive NDI is a consequence of improper AQP2 routing in the intact 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