title: Claudin-16 affects transcellular Cl- secretion in MDCK cells.
abstract: Claudin-16 (paracellin-1) is a tight junction protein localized mainly in the thick ascending limb of Henle's loop and also in the distal nephron. Its defect causes familial hypomagnesaemia with hypercalciuria and nephrocalcinosis. This had been taken as an indication that claudin-16 conveys paracellular Mg(2+) and Ca(2+) transport; however, evidence is still conflicting. We studied paracellular ion permeabilities as well as effects of claudin-16 on the driving forces for passive ion movement. MDCK-C7 cells were stably transfected with wild-type (wt) and mutant (R146T, T233R) claudin-16. Results indicated that paracellular permeability to Mg(2+) but not to Ca(2+) is increased in cells transfected with wt compared to mutant claudin-16 and control cells. Increased basolateral Mg(2+) concentration activated a transcellular Cl(-) current which was greatly enhanced in cells transfected with wt and T233R claudin-16, as compared to R146T claudin-16-transfected or control cells. This current was triggered by the basolateral calcium-sensing receptor causing Ca(2+) release from internal stores, thus activating apical Ca(2+)-sensitive Cl(-) channels and basolateral Ca(2+)-sensitive K(+) channels. Immunohistochemical data suggest that the Cl(-) channel involved is bestrophin. We conclude that claudin-16 itself possesses only moderate paracellular Mg(2+) permeability but governs transcellular Cl(-) currents by interaction with apical Ca(2+)-activated Cl(-) channels, presumably bestrophin. As the transepithelial voltage generated by such a current alters the driving force for all ions, this may be the major mechanism to regulate Mg(2+) and Ca(2+) absorption in the kidney.
authors.affiliation: Institut für Klinische Physiologie, Charité Berlin, Campus Benjamin Franklin, Hindenburgdamm 30, 12203 Berlin, Germany. dorothee.guenzel@charite.de
authors.forename: Dorothee
authors.initials: D
authors.lastname: Günzel
authors.forename: Salah
authors.initials: S
authors.lastname: Amasheh
authors.forename: Sandra
authors.initials: S
authors.lastname: Pfaffenbach
authors.forename: Jan F
authors.initials: JF
authors.lastname: Richter
authors.forename: P Jaya
authors.initials: PJ
authors.lastname: Kausalya
authors.forename: Walter
authors.initials: W
authors.lastname: Hunziker
authors.forename: Michael
authors.initials: M
authors.lastname: Fromm
mesh_terms: D000818:Animals; D001639:Bicarbonates / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D002463:Cell Membrane Permeability / Q000502:physiology; D018118:Chloride Channels / Q000378:metabolism; D002712:Chlorides / Q000378:metabolism*; D057167:Claudins; D004285:Dogs; D007668:Kidney / Q000166:cytology* / Q000378:metabolism*; D008274:Magnesium / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D011188:Potassium / Q000378:metabolism; D024681:Potassium Channels, Calcium-Activated / Q000378:metabolism; D012964:Sodium / Q000378:metabolism; D014162:Transfection
chemical_list: D001639:Bicarbonates; D018118:Chloride Channels; D002712:Chlorides; D057167:Claudins; D008565:Membrane Proteins; D024681:Potassium Channels, Calcium-Activated; C502010:claudin 16; D012964:Sodium; D008274:Magnesium; D011188:Potassium; D002118:Calcium