title: A rapid membrane potential assay to monitor CFTR function and inhibition.
abstract: The cystic fibrosis transmembrane conductance regulator (CFTR) protein is an important regulator of ion transport and fluid secretion in humans. Mutations to CFTR cause cystic fibrosis, which is a common recessive genetic disorder in Caucasians. Involvement of CFTR has been noted in other important diseases, such as secretory diarrhea and polycystic kidney disease. The assays to monitor CFTR function that have been described to date either are complicated or require specialized instrumentation and training for execution. In this report, we describe a rapid FlexStation-based membrane potential assay to monitor CFTR function. In this assay, agonist-mediated activation of CFTR results in membrane depolarization that can be monitored using a fluorescent membrane potential probe. Availability of a simple mix-and-read assay to monitor the function of this important protein might accelerate the discovery of CFTR ligands to study a variety of conditions.
authors: 1RTI International, Research Triangle Park, NC, USA.
authors: Rangan
authors: R
authors: Maitra
authors: Perumal
authors: P
authors: Sivashanmugam
authors: Keith
authors: K
authors: Warner
mesh_terms: D001681:Biological Assay; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000819:agonists* / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D005456:Fluorescent Dyes; D057809:HEK293 Cells; D006801:Humans; D017136:Ion Transport / Q000187:drug effects; D008024:Ligands; D008564:Membrane Potentials / Q000187:drug effects*; D009154:Mutation; D059629:Signal-To-Noise Ratio; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*; D013050:Spectrometry, Fluorescence; D013997:Time Factors
chemical_list: C505032:CFTR protein, human; D005456:Fluorescent Dyes; D008024:Ligands; D054852:Small Molecule Libraries; D019005:Cystic Fibrosis Transmembrane Conductance Regulator
keywords: CFTR; assay; cystic fibrosis; ion channel; membrane potential