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

Robert P. Wersto, Eugene R. Rosenthal, Ronald Crystal, Kenneth R. Spring

Research output: Contribution to journalArticle

17 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)1167-1172
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume93
Issue number3
DOIs
Publication statusPublished - 6 Feb 1996
Externally publishedYes

Fingerprint

Cystic Fibrosis Transmembrane Conductance Regulator
Fluorescent Dyes
Epithelial Cells
Cyclic AMP
Staining and Labeling
Inborn Genetic Diseases
Retroviridae
Adenoviridae
Genetic Therapy
Permeability
Coloring Agents
Adenosine Triphosphate
Mutation
Genes
dihydrorhodamine 6G

Keywords

  • gene therapy

ASJC Scopus subject areas

  • General

Cite this

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AU - Rosenthal, Eugene R.

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