Discrimination between cystic fibrosis and CFTR-corrected epithelial cells by a membrane potential-sensitive probe

Article Article scientifique Article Discrimination between cystic fibrosis and CFTR-corrected epithelial cells by a membrane potential-sensitive probe eng 2002 author Ninin-Coclet Joëlle 28418 author Rochat Thierry 28557 author Poitry Serge 26441 author Chanson Marc 0000-0001-8104-2090 32865 181-199 10.1080/019021402753570491 11936773 0190-2148 Methods to detect functional cystic fibrosis transmembrane conductance regulator (CFTR) are needed for the assessment of new therapies in cystic fibrosis (CF). We have combined patch-clamp and fluorimetric techniques to investigate whether the fluorescent voltage-sensitive probe bis-(1,3-diethylthiobarbituic anid) trimethine oxonol (Di5BAC2(3)) discriminates between changes of membrane potential (Vm evoked by cAMP in CF and CYFT-corrected epithelial cells. About 60% of the (FTR-correrced cells increased their membrane conductance and depolarized in response to cAMP, as compared to about 20% of CF cells. CFTR was found to contribute only to a fraction of the cAMP-induced responses, as indicated by the differential effects of Cl- channel blockers. Simultaneous reocording of fluorescence (AF) and membrane potential revealed that AF detected Vm changes as small as 10 mV . The relationship between deltaF and deltaVm however, was not proportional. When a large number of cells were analyzed by digital imaging, an increase in deltaF in response to cAMP was detected in the majority of CFTR-corrected cells, but only in a small proportion of CF cells. The results indicate that the DiSBAC2(3) approach is a valid tool to compare cell populations with different proportions of cells responding to CFTR arcivation by cAMP. It cannot be used, however, for quantitative assessment of functional CFTR in individual CF cells. Cyclic AMP/pharmacology Cystic Fibrosis/pathology Cystic Fibrosis Transmembrane Conductance Regulator/physiology Epithelial Cells/physiology Fluorescent Dyes/diagnostic use Humans Image Processing, Computer-Assisted/standards Membrane Potentials/drug effects/physiology Patch-Clamp Techniques Sensitivity and Specificity Thiobarbiturates/diagnostic use Tumor Cells, Cultured Dewey 616 Dewey 618 56 Département de médecine 221 58 Département de pédiatrie, gynécologie et obstétrique 223 false 180 Mucoviscidose et jonctions GAP 229 Experimental lung research 28 3 a838bac2-8653-45f0-b500-5183afd54b69 Article (Published version) Restricted Coclet-Ninin-2002-Discrimination betwe.pdf 669063 application/pdf restrictedAccess Faculté de médecine / Section de médecine clinique / Département de médecine Faculté de médecine / Section de médecine clinique / Département de pédiatrie, gynécologie et obstétrique

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Discrimination between cystic fibrosis and CFTR-corrected epithelial cells by a membrane potential-sensitive probe	Experimental lung research			2002	596	0

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Ninin-Coclet, Joëlle