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

Overview

MeSH

  • 3T3 Cells
  • Adenoviridae
  • Animals
  • Biological Transport
  • Bronchi
  • Cell Line
  • Cystic Fibrosis
  • Endocytosis
  • Epithelium
  • Genetic Vectors
  • Humans
  • Kinetics
  • Mice
  • Microscopy, Fluorescence
  • P-Glycoprotein
  • Recombinant Proteins
  • Retroviridae
  • Rhodamines
  • Staining and Labeling
  • Thionucleotides
  • Transfection

MeSH Major

  • 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.

publication date

  • February 6, 1996

has subject area

  • 3T3 Cells
  • Adenoviridae
  • Animals
  • Biological Transport
  • Bronchi
  • Cell Line
  • Chlorides
  • Cyclic AMP
  • Cystic Fibrosis
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • Endocytosis
  • Epithelium
  • Fluorescent Dyes
  • Gene Expression
  • Genetic Vectors
  • Humans
  • Kinetics
  • Mice
  • Microscopy, Fluorescence
  • P-Glycoprotein
  • Recombinant Proteins
  • Retroviridae
  • Rhodamines
  • Staining and Labeling
  • Thionucleotides
  • Transfection

Research

keywords

  • Journal Article

Identity

Language

  • eng

PubMed Central ID

  • PMC40050

PubMed ID

  • 8577734

Additional Document Info

start page

  • 1167

end page

  • 1172

volume

  • 93

number

  • 3