title: Vasopressin-stimulated chloride transport in transimmortalized mouse cell lines derived from the distal convoluted tubule and cortical and inner medullary collecting ducts.
abstract: BACKGROUND
  The fine control of NaCl absorption takes place in the distal parts of the renal tubule, but the regulation of Cl(-) transport in this region has not been fully elucidated. We have analysed the effects of dD-arginine vasopressin (dDAVP) on Cl(-) fluxes in cultured mouse distal convoluted tubule (mpkDCT), cortical collecting duct (mpkCCD) and inner medullary collecting duct (mpkIMCD) cell lines.
  
  
  METHODS
  RT-PCR and Western blotting were used to detect the amiloride-sensitive sodium channel (ENaC) and cystic fibrosis transmembrane conductance regulator (CFTR) mRNAs and protein in cultured mpkDCT, mpkCCD and mpkIMCD cells. Cl(-) fluxes were analysed by measuring the short-circuit current (I(sc)) and bidirectional (36)Cl(-) fluxes on confluent cells grown on filters.
  
  
  RESULTS
  All three cell lines expressed ENaC and CFTR and had I(sc) stimulated by dDAVP. The rise in I(sc) caused by dDAVP (10(-8) M) was inhibited by amiloride, and to a lesser extent by 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB) in all three cell lines. The dDAVP-dependent I(sc) measured under apical Na(+)-free condition was reduced by Cl(-) channel blockers with a profile (NPPB>glibenclamide>DIDS), similar to that for rat CFTR. dDAVP stimulated the apical-to-basal (36)Cl(-) flux and to a lesser extent the basal-to-apical (36)Cl(-) flux under open-circuit condition in all three cultured cell lines. Adding NPPB to the apical side reduced the basal-to-apical (36)Cl(-) flux but not the opposite (36)Cl(-) flux from dDAVP-treated cells.
  
  
  CONCLUSIONS
  These results indicate that dDAVP stimulates the bi-directional flux of Cl(-), resulting in net Cl(-)absorption, in these cultured mouse distal and collecting duct cells. I(sc) experiments also suggest the presence of a minor component of electrogenic Cl(-) secretion, possibly mediated by CFTR.
authors.affiliation: Institut National de la Santé et de la Recherche Médicale, Unité 478, Institut Fédératif de Recherche 02, Faculté de Médecine Xavier Bichat, Paris, France.
authors.forename: J P
authors.initials: JP
authors.lastname: Duong Van Huyen
authors.forename: M
authors.initials: M
authors.lastname: Bens
authors.forename: J
authors.initials: J
authors.lastname: Teulon
authors.forename: A
authors.initials: A
authors.lastname: Vandewalle
mesh_terms: D000818:Animals; D027321:Anion Transport Proteins; D001127:Arginine Vasopressin / Q000502:physiology*; D001692:Biological Transport / Q000187:drug effects; D002352:Carrier Proteins / Q000502:physiology; D002461:Cell Line, Transformed; D002712:Chlorides / Q000493:pharmacokinetics*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics / Q000378:metabolism; D003894:Deamino Arginine Vasopressin / Q000494:pharmacology; D004594:Electrophysiology; D053503:Epithelial Sodium Channels; D007672:Kidney Cortex; D007679:Kidney Medulla; D007685:Kidney Tubules, Collecting / Q000166:cytology / Q000378:metabolism* / Q000502:physiology; D007686:Kidney Tubules, Distal / Q000166:cytology / Q000378:metabolism* / Q000502:physiology; D051379:Mice; D008822:Mice, Transgenic; D012333:RNA, Messenger / Q000378:metabolism; D012076:Renal Agents / Q000494:pharmacology; D015222:Sodium Channels / Q000235:genetics / Q000378:metabolism
chemical_list: D027321:Anion Transport Proteins; D002352:Carrier Proteins; D002712:Chlorides; D053503:Epithelial Sodium Channels; D012333:RNA, Messenger; D012076:Renal Agents; D015222:Sodium Channels; D001127:Arginine Vasopressin; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D003894:Deamino Arginine Vasopressin