Permeabilization via the P2X7 purinoreceptor reveals the presence of a Ca2+-activated Cl- conductance in the apical membrane of murine tracheal epithelial cells

被引:27
作者
Gabriel, SE [1 ]
Makhlina, M
Martsen, E
Thomas, EJ
Lethem, MI
Boucher, RC
机构
[1] Univ N Carolina, CFPRT Ctr, Chapel Hill, NC 27599 USA
[2] Univ N Carolina, Cyst Fibrosis Pulm Res & Clin Treatment Ctr, Chapel Hill, NC 27599 USA
[3] Univ Brighton, Sch Pharm & Biomol Sci, Brighton BN2 4GJ, E Sussex, England
关键词
D O I
10.1074/jbc.M004953200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Calcium-activated Cl- secretion is an important modulator of regulated ion transport in murine airway epithelium and is mediated by an unidentified Ca2+-stimulated Cl- channel. We have transfected immortalized murine tracheal epithelial cells with the cDNA encoding the permeabilizing P2X(7) purinoreceptor (P2X(7)-R) to selectively permeabilize the basolateral membrane and thereby isolate the apical membrane Ca2+-activated Cl- current. In P2X(7)-R-permeabilized cells, we have demonstrated that UTP stimulates a Cl- current across the apical membrane of CF and normal murine tracheal epithelial cells. The magnitude of the UTP-stimulated current was significantly greater in CF than in normal cells. Ion substitution studies demonstrated that the current exhibited a permselectivity sequence of Cl- > I- > Br- > gluconate(-). We have also determined a rank order of potency for putative Cl- channel blockers: niflumic acid greater than or equal to 5-nitro-2-(3-phenylpropylamino)benzoic acid > 4,4'-diisothiocyanostilbene-2,2'-disulfonate > glybenclamide much greater than diphenlyamine-2-carboxylate, tamoxifen, and p-tetra-sulfonato-tetra-methoxy-calix[4]arene. Complete characterization of this current and the corresponding single channel properties could lead to the development of a new therapy to correct the defective airway surface liquid in cystic fibrosis patients.
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页码:35028 / 35033
页数:6
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