Localization of cystic fibrosis transmembrane conductance regulator in chloride secretory epithelia.

作者：Gerene M. Denning, Lynda S. Ostedgaard, Shuk Han Cheng, Alan Edward Smith, Michael J. Welsh · 发表于：Journal of Clinical Investigation · 年份：1992 · DOI：10.1172/jci115582 · 被引用次数：241 · 研究领域：Cystic Fibrosis Research Advances、Neonatal Respiratory Health Research、Bacterial biofilms and quorum sensing

Cystic fibrosis (CF),' the most common lethal genetic disease in Caucasians (1), is caused by mutations in the gene for the cystic fibrosis transmembrane conductance regulator (CFTR) (2-4).Early studies on the physiologic basis ofthe disease dem- onstrated that epithelia in CF patients are less permeable to Cl-.Subsequent studies showed that this reduced permeability to Clwas due to a lack ofcAMP-regulated Cl-channels in the apical membrane ofsecretory epithelia (5).Recent studies indi- cate that CFTR itself is a cAMP-regulated Cl-channel (6-8) that is expressed primarily in epithelia (2), results that directly link mutations in CFTR to reduced Cl-permeability in CF epithelia.For CFTR chloride channels to govern transepithe- lial Cl-secretion, they must be located in the apical membrane.We used confocal immunofluorescence microscopy to test the hypothesis that CFTR is located in the apical membrane of Cl--secreting epithelial cells. Methods Materials. L-[35S]methionine (1,000Ci/mmol), fish gelatin, normal goat serum, and FITC-conjugated streptavidin were obtained from Amersham Corp. (Arlington Heights, IL).Biotinylated secondary anti- bodies were purchased from Vector Laboratories, Inc. (Burlingame, CA) or Amersham Corp. FHTC-conjugated wheat germ agglutinin and [6-((6-((biotinoyl)amino)hexanoyl)amino) hexanoic acid, succinimi- dyl ester] (biotin-XX) were purchased from Molecular Probes, Inc. (Eugene, OR).Rabbit anti-human placental alkaline phosphatase and FHTC-conjugated ra...