Uptake of fluorescent dyes associated with the functional expression of the cystic fibrosis transmembrane conductance regulator in epithelial cells.

Publication Type Academic Article
Authors Wersto R, Rosenthal E, Crystal R, Spring K
Journal Proc Natl Acad Sci U S A
Volume 93
Issue 3
Pagination 1167-72
Date Published 02/06/1996
ISSN 0027-8424
Keywords Chlorides, Cyclic AMP, Cystic Fibrosis Transmembrane Conductance Regulator, Fluorescent Dyes, Gene Expression
Abstract Specific mutations in the cystic fibrosis transmembrane conductance regulator (CFTR), the most common autosomal recessive fatal genetic disease of Caucasians, result in the loss of epithelial cell adenosine 3',5'-cyclic-monophosphate (cAMP)-stimulated Cl- conductance. We show that the influx of a fluorescent dye, dihydrorhodamine 6G (dR6G), is increased in cells expressing human CFTR after retrovirus- and adenovirus-mediated gene transfer. dR6G influx is stimulated by cAMP and is inhibited by antagonists of cAMP action. Dye uptake is ATP-dependent and inhibited by Cl- removal or the addition of 10 mM SCN-. Increased staining is associated with functional activation of CFTR Cl- permeability. dR6G staining enables both the fluorescent assessment of CFTR function and the identification of successfully corrected cells after gene therapy.
DOI 10.1073/pnas.93.3.1167
PubMed ID 8577734
PubMed Central ID PMC40050
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