Methods to monitor cell surface expression and endocytic trafficking of CFTR in polarized epithelial cells

Methods Mol Biol. 2011:741:271-83. doi: 10.1007/978-1-61779-117-8_18.

Abstract

Cystic fibrosis transmembrane conductance regulator (CFTR)-mediated chloride secretion is critical to maintaining airway surface hydration and efficient mucociliary clearance in the upper airways. Mutations in CFTR in cystic fibrosis lead to reduced expression of functional CFTR channels at the apical plasma membrane of the airway epithelium, leading to dehydration of the airway surface liquid and diminished mucociliary clearance. Cell surface CFTR is modulated by changes in CFTR endocytosis and recycling, effectively altering the cell surface abundance of the channel. This chapter examines current methods employed to measure the cell surface expression of CFTR, as well as methods to monitor CFTR movement through the endocytic pathway.

MeSH terms

  • Biotinylation
  • Cell Polarity*
  • Cystic Fibrosis Transmembrane Conductance Regulator / metabolism*
  • Cytological Techniques / methods*
  • Endocytosis*
  • Epithelial Cells / cytology*
  • Epithelial Cells / metabolism*
  • Gene Expression Regulation*
  • Humans
  • Intracellular Space / metabolism

Substances

  • Cystic Fibrosis Transmembrane Conductance Regulator