title: Extracellular GTP is a potent water-transport regulator via aquaporin 5 plasma-membrane insertion in M1-CCD epithelial cortical collecting duct cells.
abstract: OBJECTIVE
  Extracellular GTP is able to modulate some specific functions in neuron, glia and muscle cell models as it has been demonstrated over the last two decades. In fact, extracellular GTP binds its specific plasma membrane binding sites and induces signal transduction via [Ca(2+)]i increase. We demonstrate, for the first time, that extracellular GTP is able to modulate cell swelling in M1-CCD cortical collecting duct epithelial cells via upregulation of aquaporin 5 (AQP5) expression.
  
  
  METHODS
  We used videoimaging, immunocitochemistry, flow cytometry, confocal techniques, Western blotting and RT-PCR for protein and gene expression analysis, respectively.
  
  
  RESULTS
  We demonstrate that AQP5 mRNA is up-regulated 7 h after the GTP exposure in the cell culture medium, and its protein level is increased after 12-24 h. We show that AQP5 is targeted to the plasma membrane of M1-CCD cells, where it facilitates cell swelling, and that the GTP-dependent AQP5 up-regulation occurs via [Ca(2+)]i increase. Indeed, GTP induces both oscillating and transient [Ca(2+)]i increase, and specifically the oscillating kinetic appears to be responsible for blocking cell cycle in the S-phase while the [Ca(2+)]i influx, with whatever kinetic, seems to be responsible for inducing AQP5 expression.
  
  
  CONCLUSIONS
  The role of GTP as a regulator of AQP5-mediated water transport in renal cells is of great importance in the physiology of renal epithelia, due to its possible physiopathological implications. GTP-dependent AQP5 expression could act as osmosensor. In addition, the data presented here suggest that GTP might play the same role in other tissues where rapid water transport is required for cell volume regulation and maintenance of the homeostasis.
authors.affiliation: Department of Neurosciences and Imaging (DNI), 'G. d'Annunzio' University of Chieti-Pescara, Chieti, Italy.
authors.forename: Rosa
authors.initials: R
authors.lastname: Mancinelli
authors.forename: Rita Maria Laura
authors.initials: RM
authors.lastname: La Rovere
authors.forename: Stefania
authors.initials: S
authors.lastname: Fulle
authors.forename: Sebastiano
authors.initials: S
authors.lastname: Miscia
authors.forename: Marco
authors.initials: M
authors.lastname: Marchisio
authors.forename: Laura
authors.initials: L
authors.lastname: Pierdomenico
authors.forename: Paola
authors.initials: P
authors.lastname: Lanuti
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Procino
authors.forename: Claudia
authors.initials: C
authors.lastname: Barbieri
authors.forename: Maria
authors.initials: M
authors.lastname: Svelto
authors.forename: Giorgio
authors.initials: G
authors.lastname: Fanò-Illic
authors.forename: Tiziana
authors.initials: T
authors.lastname: Pietrangelo
mesh_terms: D000818:Animals; D051402:Aquaporin 5 / Q000096:biosynthesis*; D020013:Calcium Signaling / Q000187:drug effects; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000494:pharmacology*; D007685:Kidney Tubules, Collecting / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D015854:Up-Regulation / Q000187:drug effects*
chemical_list: C496038:Aqp5 protein, mouse; D051402:Aquaporin 5; D006160:Guanosine Triphosphate