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PMID:17596272

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Citation

Robert, R, Savineau, JP, Norez, C, Becq, F and Guibert, C (2007) Expression and function of cystic fibrosis transmembrane conductance regulator in rat intrapulmonary arteries. Eur. Respir. J. 30:857-64

Abstract

The cystic fibrosis transmembrane conductance regulator (CFTR) gene encodes a cyclic adenosine monophosphate (cAMP)-dependent chloride channel located mainly at the apical membrane of epithelial cells. In myocytes of pulmonary arteries, numerous chloride channels have been identified and described, but not the CFTR. Thus the presence and function of the CFTR was investigated in rat intrapulmonary arteries. CFTR expression, localisation and function were analysed in cultured smooth muscle cells using Reverse transcriptase (RT)-PCR and immunoprecipitation followed by protein kinase A phosphorylation, immunolocalisation and an iodide efflux assay, respectively. The role of the CFTR in pulmonary vasoreactivity was determined in arterial rings using an organ bath system. RT-PCR and immunoprecipitation analyses, as well as the immunolocalisation study, revealed the expression of CFTR gene transcripts and protein. The iodide efflux assay showed the existence of functional cAMP-, calcium- and volume-dependent chloride channels. Furthermore, the following effects were found: 1) inhibition of forskolin/genistein-activated iodide efflux by glibenclamide, diphenylamine-2-carboxylic acid and CFTR-specific inhibitor (CFTR(inh))-172; 2) activation of iodide efflux by the benzoquinolizinium derivative CFTR activators MPB-07 and MPB-91; and 3) inhibition of MPB-dependent efflux by CFTR(inh)-172. Finally, CFTR activators induced concentration-dependent vasorelaxation in rings preconstricted with phenylephrine, in the presence or absence of endothelium. The present results are the first to reveal functional cyclic adenosine monophosphate-regulated cystic fibrosis transmembrane conductance regulator contributing to endothelium-independent vasorelaxation in rat intrapulmonary arterial myocytes.

Links

PubMed Online version:10.1183/09031936.00060007

Keywords

Analysis of Variance; Animals; Cells, Cultured; Chloride Channels/metabolism; Chloride Channels/physiology; Cyclic AMP/pharmacology; Cystic Fibrosis Transmembrane Conductance Regulator/metabolism; Cystic Fibrosis Transmembrane Conductance Regulator/physiology; Forskolin/pharmacology; Genistein/pharmacology; Glyburide/pharmacology; Immunoprecipitation; Iodides/metabolism; Male; Muscle, Smooth, Vascular/metabolism; Muscle, Smooth, Vascular/physiology; Phosphorylation; Pulmonary Artery; Rats; Rats, Wistar; Reverse Transcriptase Polymerase Chain Reaction; Vasodilation/physiology

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

RAT:CFTR

involved_in

GO:0042311: vasodilation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

RAT:CFTR

involved_in

GO:0015705: iodide transport

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

RAT:CFTR

involved_in

GO:0097755: positive regulation of blood vessel diameter

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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