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 |