title: Aquaporin expression contributes to human transurothelial permeability in vitro and is modulated by NaCl.
abstract: It is generally considered that the bladder is impervious and stores urine in unmodified form on account of the barrier imposed by the highly-specialised uro-epithelial lining. However, recent evidence, including demonstration of aquaporin (AQP) expression by human urothelium, suggests that urothelium may be able to modify urine content. Here we have we applied functional assays to an in vitro-differentiated normal human urothelial cell culture system and examined both whether AQP expression was responsive to changes in osmolality, and the effects of blocking AQP channels on water and urea transport. AQP3 expression was up-regulated by increased osmolality, but only in response to NaCl. A small but similar effect was seen with AQP9, but not AQP4 or AQP7. Differentiated urothelium revealed significant barrier function (mean TER 3862 Ω.cm(2)), with mean diffusive water and urea permeability coefficients of 6.33×10(-5) and 2.45×10(-5) cm/s, respectively. AQP blockade with mercuric chloride resulted in decreased water and urea flux. The diffusive permeability of urothelial cell sheets remained constant following conditioning in hyperosmotic NaCl, but there was a significant increase in water and urea flux across an osmotic gradient. Taken collectively with evidence emerging from studies in other species, our results support an active role for human urothelium in sensing and responding to hypertonic salt concentrations through alterations in AQP protein expression, with AQP channels providing a mechanism for modifying urine composition. These observations challenge the traditional concept of an impermeable bladder epithelium and suggest that the urothelium may play a modulatory role in water and salt homeostasis.
authors.affiliation: Jack Birch Unit for Molecular Carcinogenesis, Department of Biology, University of York, York, United Kingdom.
authors.forename: Peter C
authors.initials: PC
authors.lastname: Rubenwolf
authors.forename: Nikolaos T
authors.initials: NT
authors.lastname: Georgopoulos
authors.forename: Lisa A
authors.initials: LA
authors.lastname: Kirkwood
authors.forename: Simon C
authors.initials: SC
authors.lastname: Baker
authors.forename: Jennifer
authors.initials: J
authors.lastname: Southgate
mesh_terms: D051400:Aquaporin 3 / Q000235:genetics / Q000378:metabolism; D051401:Aquaporin 4 / Q000235:genetics / Q000378:metabolism; D020346:Aquaporins / Q000235:genetics / Q000378:metabolism*; D001692:Biological Transport; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D006801:Humans; D008856:Microscopy, Fluorescence; D009994:Osmolar Concentration; D012965:Sodium Chloride / Q000494:pharmacology*; D014508:Urea / Q000378:metabolism; D019459:Urothelium / Q000166:cytology* / Q000187:drug effects; D014867:Water / Q000378:metabolism
chemical_list: C496027:AQP3 protein, human; C114141:AQP7 protein, human; C109669:AQP9 protein, human; D051401:Aquaporin 4; D020346:Aquaporins; D014867:Water; D051400:Aquaporin 3; D012965:Sodium Chloride; D014508:Urea