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• W2082646413 abstract "The cystic fibrosis transmembrane conductance regulator (CFTR) [[1]Riordan J.R. Rommens J.M. Kerem B-S. Alon N. Rozmahel R. Grzelczak Z. et al.Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA.Science. 1989; 245: 1066-1073Crossref PubMed Scopus (5925) Google Scholar] is a unique member of the ATP-binding cassette (ABC) transporter family that plays a central role in salt and liquid movements across epithelial tissues [[2]Welsh M.J. Ramsey B.W. Accurso F. Cutting G.R. Cystic fibrosis.in: Scriver C.R. Beaudet A.L. Sly W.S. Valle D. The Metabolic and Molecular Basis of Inherited Disease. McGraw-Hill, New York2001: 5121-5188Google Scholar]. CFTR is a multi-functional protein: its best characterised function is as a Cl− channel with complex regulation [3Gadsby D.C. Nairn A.C. Control of cystic fibrosis transmembrane conductance regulator channel gating by phosphorylation and nucleotide hydrolysis.Physiol. Rev. 1999; 79: S77-S107PubMed Google Scholar, 4Sheppard D.N. Welsh M.J. Structure and function of the cystic fibrosis transmembrane conductance regulator chloride channel.Physiol. Rev. 1999; 79: S23-S45PubMed Google Scholar]. In addition, CFTR has been demonstrated to regulate the activity of other ion channels and transporters in epithelial cells [[5]Schwiebert E.M. Benos D.J. Egan M.E. Stutts M.J. Guggino W.B. CFTR is a conductance regulator as well as a chloride channel.Physiol. Rev. 1999; 79: S145-S166PubMed Google Scholar]. Loss of these different functions of CFTR disrupts transepithelial electrolyte transport in a variety of organs to cause the wide-ranging manifestations characteristic of cystic fibrosis (CF) [[2]Welsh M.J. Ramsey B.W. Accurso F. Cutting G.R. Cystic fibrosis.in: Scriver C.R. Beaudet A.L. Sly W.S. Valle D. The Metabolic and Molecular Basis of Inherited Disease. McGraw-Hill, New York2001: 5121-5188Google Scholar]. The Cell Physiology Section of the Online Virtual Repository of Methods and Reagents for CFTR Expression and Functional Studies (Section D) [[6]European Working Group on CFTR Expression. The Online Virtual Repository of Methods and Reagents for CFTR Expression and Functional Studies (section D). 2004. http://pen2.igc.gulbenkian.pt/cftr/vr/physiology.html.Google Scholar] contains consensus protocols on many of the different approaches that researchers employ to understand better CFTR and its dysfunction in disease. It also provides a catalogue of pharmacological reagents that can be used to modulate CFTR Cl− currents. These protocols are not intended to be step-by-step instructions for researchers to follow when using specific techniques to study CFTR. Instead, their purpose is to provide valuable help and advice to novice and expert alike as they seek new knowledge and understanding of CFTR. To investigate the different functions of CFTR, a rich variety of materials and methods are used. The activity of wild-type and mutant CFTR is studied in native tissues and heterologous expression systems at the molecular, cellular and tissue levels. Many of the methods are electrophysiological ranging from high-resolution recording of individual CFTR Cl− channels using the patch-clamp technique [[7]Sheppard D.N. Gray M.A. Gong X. Sohma Y. Kogan I. Benos D.J. et al.The patch-clamp and planar lipid bilayer techniques: powerful tools to investigate the CFTR Cl− channel.J. Cystic. Fibrosis. 2004; 3: 101-108Abstract Full Text Full Text PDF PubMed Scopus (25) ] to Cl− flow across polarised epithelia monitored with the Ussing chamber technique [[8]Li H. Sheppard D.N. Hug M.J. Transepithelial electrical measurements with the Ussing chamber.J. Cystic. Fibrosis. 2004; 3: 123-126Abstract Full Text Full Text PDF PubMed Scopus (108) ]. Other methods employ planar lipid bilayers and ATPases assays to understand, at the molecular level, how ATP hydrolysis drives conformation changes in the CFTR Cl− channel [7Sheppard D.N. Gray M.A. Gong X. Sohma Y. Kogan I. Benos D.J. et al.The patch-clamp and planar lipid bilayer techniques: powerful tools to investigate the CFTR Cl− channel.J. Cystic. Fibrosis. 2004; 3: 101-108Abstract Full Text Full Text PDF PubMed Scopus (25) , 9Kogan I. Ramjeesingh M. Bear C.E. ATPase assay of purified, reconstituted CFTR protein.J. Cystic. Fibrosis. 2004; 3: 133-134Abstract Full Text Full Text PDF PubMed Scopus (3) ]. Yet other methods employ fluorescent dyes and radioisotopes to assay the function of CFTR in populations of cells [10Munkonge F. Alton E.W.F.W. Andersson C. Davidson H. Dragomir A. Edelman A. et al.Measurement of halide efflux from cultured and primary airway epithelial cells using fluorescent indicators.J. Cystic. Fibrosis. 2004; 3: 171-176Abstract Full Text Full Text PDF PubMed Scopus (47) , 11Norez C. Heda G.D. Jensen T. Kogan I. Hughes L.K. Auzanneau C. et al.Determination of CFTR chloride channel activity and pharmacology using radiotracer flux methods.J. Cystic. Fibrosis. 2004; 3: 119-121Abstract Full Text Full Text PDF PubMed Scopus (40) ]. Finally, innovative techniques using confocal and electron microscopy have been developed to investigate the pathogenesis of CF lung disease [[12]Roomans G.M. Kozlova I. Nilsson H. Vanthanouvong V. Button B. Tarran R. Measurement of airway surface liquid height and mucus transport by fluorescence microscopy, and of ion composition by X-ray microanalysis.J. Cystic. Fibrosis. 2004; 3: 135-139Abstract Full Text Full Text PDF PubMed Scopus (16) ]. The following articles provide an overview of the rich variety of protocols in the Cell Physiology Section of the Online Virtual Repository of Methods and Reagents for CFTR Expression and Functional Studies [[6]European Working Group on CFTR Expression. The Online Virtual Repository of Methods and Reagents for CFTR Expression and Functional Studies (section D). 2004. http://pen2.igc.gulbenkian.pt/cftr/vr/physiology.html.Google Scholar] compiled by the European Working Group on CFTR Expression. We encourage you strongly to visit the Online Virtual Repository and examine in detail the protocols. doi:10.1016/j.jcf.2004.05.046 doi:10.1016/j.jcf.2004.05.025 doi:10.1016/j.jcf.2004.05.026 doi:10.1016/j.jcf.2004.05.036 doi:10.1016/j.jcf.2004.05.025 doi:10.1016/j.jcf.2004.05.029" @default.
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• W2082646413 title "Section D (cell physiology): an armamentarium to investigate CFTR function" @default.
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