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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
40
pubmed:dateCreated
1996-11-25
pubmed:abstractText
In this study, we describe the establishment of a stably transfected epithelial cell line with the cDNA for the rat aquaporin 2 (AQP2). To this end, we used a human cell line (HCD) derived from the cortical collecting duct and having characteristics of principal cells (Prié, D., Friedlander, G., Coureau, C., Vandewalle, A., Cassigena, R., and Ronco, P. M. (1995) Kidney Int. 47, 1310-1318). The HCD cells were first screened for the constitutive expression of AQPs. By Western blot analysis, we found a low expression of immunoreactive AQP2 and AQP4 proteins. In contrast, transfected cells (clone CD8) probed with AQP2 antiserum expressed an intense 29-kDa protein on immunoblot in addition to a broad band between 35-45 kDa corresponding to the glycosylated form of the protein, indicating that full maturity of the protein is attained in transfected cells. Immunofluorescence demonstrated that AQP2 was located in intracellular vesicles. After vasopressin stimulation, the staining redistributed from an intracellular site to the apical pole of the cells, an effect similar to that described on collecting duct principal cells in vivo (Sabolic, I., Katsura, T., Verbavatz, J. M., and Brown, D. (1995) J. Membr. Biol. 143, 165-175) and in perfused tubules (Nielsen, S., Chou, C. L., Marples, D., Christensen, E. I., Kishore, B. K., and Knepper, M. A. (1995) Proc. Natl. Acad. Sci. U. S. A. 92, 1013-1017). The redistribution of AQP2 in CD8 cells was accompanied by an approximately 6-fold increase in osmotic water permeability coefficient (Pf), which was inhibited by 0.3 m HgCl2. These data indicate that functional vasopressin-sensitive water channels are expressed in transfected cells. The stably transfected cells represent a suitable model to unravel by direct experimental approach the intracellular signals involved in the translocation of AQP2 to the apical plasma membrane in the presence of vasopressin.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
4
pubmed:volume
271
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
24365-70
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed:year
1996
pubmed:articleTitle
Expression and functional analysis of water channels in a stably AQP2-transfected human collecting duct cell line.
pubmed:affiliation
Istituto di Fisiologia Generale, Università degli Studi di Bari, Via Amendola 165/A, 70126 Bari, Italy.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't