Adenovirus-mediated expression of aquaporin-5 in epithelial cells.
A recombinant adenovirus coding for rat aquaporin-5 was constructed and plaque purified. The recombinant adenovirus (AdrAQP5) mediated the expression of aquaporin-5 in rat and human salivary cell lines and in dog kidney cells in vitro as demonstrated by Northern blot and Western blot analyses, and by confocal microscopy after immunofluorescent labeling. In kidney cells, expression of the transgene was optimal if cells were infected at their basolateral surface, a phenomenon associated with the distribution of integrin receptors on these cells. The expressed aquaporin-5 protein was functionally active because viral-mediated gene transfer resulted in a significant increase in the osmotically directed net fluid secretion rate across monolayers of kidney cells. AdrAQP5 should provide an efficient and useful means to impart facilitated water permeability to cells lacking such a pathway.
Delporte
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Clinical Investigations and Patient Care Branch, NIDR, National Institutes of Health, Bethesda, Maryland 20892-1190, USA.
O'Connell
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He
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Ambudkar
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Agre
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Baum
B J
BJ
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D051402:Aquaporin 5; D020346:Aquaporins; D015152:Blotting, Northern; D015153:Blotting, Western; D002874:Chromosome Mapping; D004285:Dogs; D018014:Gene Transfer Techniques; D006801:Humans; D016023:Integrins / Q000032:analysis; D007473:Ion Channels / Q000096:biosynthesis* / Q000235:genetics; D007668:Kidney / Q000378:metabolism; D008565:Membrane Proteins; D018613:Microscopy, Confocal; D051381:Rats; D012469:Salivary Glands / Q000378:metabolism*; D014018:Tissue Distribution
C496036:AQP5 protein, human; C496040:Aqp5 protein, rat; D051402:Aquaporin 5; D020346:Aquaporins; D016023:Integrins; D007473:Ion Channels; D008565:Membrane Proteins