Gain-of-function mutations of the V2 vasopressin receptor in nephrogenic syndrome of inappropriate antidiuresis (NSIAD): a cell-based assay to assess constitutive water reabsorption

Marianna Ranieri, Grazia Tamma, Tommaso Pellegrino, Vanessa Vezzi, Caterina Ambrosio, Cristina Grò, Annarita Di Mise, Tommaso Costa, Giovanna Valenti, Susanna Cotecchia

Research output: Contribution to journalArticlepeer-review

Abstract

Nephrogenic syndrome of inappropriate antidiuresis (NSIAD) is a recently identified chromosome X-linked disease associated with gain-of-function mutations of the V2 vasopressin receptor (V2R), a G-protein-coupled receptor. It is characterized by inability to excrete a free water load, hyponatremia, and undetectable vasopressin-circulating levels. Hyponatremia can be quite severe in affected male children. To gain a deeper insight into the functional properties of the V2R active mutants and how they might translate into the pathological outcome of NSIAD, in this study, we have expressed the wild-type V2R and three constitutively active V2R mutants associated with NSIAD (R137L, R137C, and the F229V) in MCD4 cells, a cell line derived from renal mouse collecting duct, stably expressing the vasopressin-sensitive water channel aquaporin-2 (AQP2). Our findings indicate that in cells expressing each active mutant, AQP2 was constitutively localized to the apical plasma membrane in the absence of vasopressin stimulation. In line with these observations, under basal conditions, osmotic water permeability in cells expressing the constitutively active mutants was significantly higher compared to that of cells expressing the wild-type V2R. Our findings demonstrate a direct link between activating mutations of the V2R and the perturbation of water balance in NSIAD. In addition, this study provides a useful cell-based assay system to assess the functional consequences of newly discovered activating mutations of the V2R on water permeability in kidney cells and to screen the effect of drugs on the mutated receptors.

Original languageEnglish
Pages (from-to)1291-1304
Number of pages14
JournalPflugers Archiv European Journal of Physiology
Volume471
Issue number10
DOIs
Publication statusPublished - Oct 2019

Keywords

  • Animals
  • Aquaporin 2/metabolism
  • Cell Line
  • Gain of Function Mutation
  • Genetic Diseases, X-Linked/genetics
  • Humans
  • Inappropriate ADH Syndrome/genetics
  • Mice
  • Receptors, Vasopressin/genetics
  • Renal Reabsorption
  • Vasopressins/metabolism
  • Water/metabolism
  • Water-Electrolyte Balance

Fingerprint Dive into the research topics of 'Gain-of-function mutations of the V2 vasopressin receptor in nephrogenic syndrome of inappropriate antidiuresis (NSIAD): a cell-based assay to assess constitutive water reabsorption'. Together they form a unique fingerprint.

Cite this