Document Detail


A method for measuring Cl efflux from dispersed cells of airway epithelium.
MedLine Citation:
PMID:  7539784     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
This paper describes a method for measuring the increase in halide permeability of isolated airway epithelial cells induced by adenosine 3',5'-cyclic monophosphate (cAMP). Suspensions of isolated cells, known to contain the cystic fibrosis transmembrane conductance regulator (CFTR), were placed in the upper part of a Swinnex filter holder containing a filter with pores of 0.65 micron diameter. Medium was perfused over the cells at room temperature and collected at minute intervals following its passage through the filter. Experiments were performed on Calu-3 and T84 cells (human lung and colonic epithelial cell lines), primary cultures of dog and human tracheal epithelium, and Swiss 3T3 fibroblasts stably transfected with CFTR. In all cell types, addition of agents that elevate cAMP led to increases in the rates of loss of 36Cl and 125I. However, in human tracheal epithelial cells, warming the medium from room temperature to 37 degrees C was a more effective way of stimulating tracer efflux. Increases in efflux in response to either temperature or cAMP-elevating agents were inhibited by diphenylamine-2-carboxylate, a blocker of CFTR. Reproducible increases in tracer efflux were seen with as few as 10(6) cells. Cells that had been trypsinized off their culture dishes responded better than cells that had been scraped off, although treatment of scraped cells with trypsin enhanced their responsiveness to cAMP-elevating agents. Cystic fibrosis is characterized by the lack of a cAMP-activated Cl conductance in the apical membrane of airway epithlia.(ABSTRACT TRUNCATED AT 250 WORDS)
Authors:
T Ohrui; B Q Shen; R J Mrsny; J H Widdicombe
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Journal of applied physiology (Bethesda, Md. : 1985)     Volume:  78     ISSN:  8750-7587     ISO Abbreviation:  J. Appl. Physiol.     Publication Date:  1995 Mar 
Date Detail:
Created Date:  1995-07-11     Completed Date:  1995-07-11     Revised Date:  2013-09-26    
Medline Journal Info:
Nlm Unique ID:  8502536     Medline TA:  J Appl Physiol (1985)     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1197-202     Citation Subset:  IM    
Affiliation:
Cystic Fibrosis Research Center, University of California, San Francisco 94143, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Membrane Permeability
Chlorides / metabolism*
Chlorine
Cyclic AMP / physiology
Cystic Fibrosis Transmembrane Conductance Regulator
Dogs
Electrophysiology
Epithelial Cells
Epithelium / metabolism,  physiology
Humans
Iodine Radioisotopes
Membrane Proteins / metabolism
Physiology / instrumentation,  methods*
Radioisotopes
Trachea / cytology,  metabolism*,  physiology
Grant Support
ID/Acronym/Agency:
DK-47766/DK/NIDDK NIH HHS; HL-42368/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/CFTR protein, human; 0/Chlorides; 0/Iodine Radioisotopes; 0/Membrane Proteins; 0/Radioisotopes; 126880-72-6/Cystic Fibrosis Transmembrane Conductance Regulator; 60-92-4/Cyclic AMP; 7782-50-5/Chlorine

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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