DIFFERENTIAL EXPRESSION OF ORCC AND CFTR INDUCED BY LOW-TEMPERATURE IN CF AIRWAY EPITHELIAL-CELLS
DIFFERENTIAL EXPRESSION OF ORCC AND CFTR INDUCED BY LOW-TEMPERATURE IN CF AIRWAY EPITHELIAL-CELLS
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DOI：
10.1152/ajpcell.1995.268.1.c243
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发表时间：
1995-01-01
影响因子：
5.5
通讯作者：
GUGGINO, WB
中科院分区：
文献类型：
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作者：
EGAN, ME;SCHWIEBERT, EM;GUGGINO, WB
当表达Delta F508-囊性纤维化跨膜传导调节因子（CFTR）突变的非上皮细胞类型在降低的温度下生长时，突变蛋白可以被适当地加工。尚未研究低温对内源性表达Delta F508-CFTR突变的气道上皮细胞中Cl-通道活性的影响。因此，我们使用由内而外和全细胞膜片钳记录、Cl-36（-）流出试验和免疫细胞化学相结合的方法，研究了孵育温度对正常、囊性纤维化（CF）受累和野生型CFTR补充CF气道上皮细胞CFTR和外向整流Cl-通道（ORCC）活性的影响。我们报告说，在25-27摄氏度的孵育CF影响气道上皮细胞与蛋白激酶A刺激CFTR样氯离子电导的外观。然而，除了CFTR Cl-通道活性的出现之外，当细胞在25-27 ℃下生长时，活性ORCC的数量减少，这表明孵育温度的降低可能与气道上皮细胞中离子通道表达和/或调节的多种改变有关。
When nonepithelial cell types expressing the Delta F508-cystic fibrosis transmembrane conductance regulator (CFTR) mutation are grown at reduced temperatures, the mutant protein can be properly processed. The effect of low temperatures on Cl- channel activity in airway epithelial cells that endogenously express the Delta F508-CFTR mutation has not been investigated. Therefore, we examined the effect of incubation temperature on both CFTR and outwardly rectifying Cl- channel (ORCC) activity in normal, in cystic fibrosis (CF)-affected, and in wild-type CFTR-complemented CF airway epithelia with use of a combination of inside-out and whole cell patch-clamp recording, Cl-36(-) efflux assays, and immunocytochemistry. We report that incubation of CF-affected airway epithelial cells at 25-27 degrees C is associated with the appearance of a protein kinase A-stimulated CFTR-like Cl- conductance. In addition to the appearance of CFTR Cl- channel activity, there is, however, a decrease in the number of active ORCC when cells are grown at 25-27 degrees C, suggesting that the decrease in incubation temperature may be associated with multiple alterations in ion channel expression and/or regulation in airway epithelial cells.