Expression and regulation of ClC-5 chloride channels: effects of antisense and oxidants.
Expression and regulation of ClC-5 chloride channels: effects of antisense and oxidants.
复制标题
ClC-5 氯离子通道的表达和调节：反义和氧化剂的影响。
DOI：
10.1152/ajpcell.2001.280.6.c1511
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
Wills,NK
中科院分区：
文献类型：
--
作者：
Weng,TX;Mo,L;Hellmich,HL;Yu,AS;Wood,T;Wills,NK
Cl−通道ClC-5的基因突变导致人类的登特氏病。我们最近从A6细胞系中克隆了一个XenopusClC-5（xClC-5）的两栖动物直系同源物。我们现在比较哺乳动物（COS-7）细胞和异种卵母细胞中表达的ClC-5电流的性质和调节。转染xClC-5 cDNA的COS-7细胞中的全细胞电流具有较强的外向整流性，Cl−> I−阴离子敏感性，并且在低pH下受到抑制，与之前在卵母细胞中的结果相似。在卵母细胞中，反义xClC-5 cRNA注射对内源性膜电流或人ClC-5的异源表达没有影响。cAMP激活剂和蛋白激酶C抑制剂对COS-7细胞或卵母细胞中表达的ClC-5电流没有显著影响，而H-89，cAMP依赖性蛋白激酶（PKA）抑制剂，和过氧化氢降低电流。我们得出结论，ClC-5电流的基本性质是独立的用于表达的宿主细胞类型。此外，ClC-5通道可能受到PKA和活性氧的调节。
Genetic mutations of the Cl−channel ClC-5 cause Dent's disease in humans. We recently cloned an amphibian ortholog ofXenopusClC-5 (xClC-5) from the A6 cell line. We now compare the properties and regulation of ClC-5 currents expressed in mammalian (COS-7) cells andXenopusoocytes. Whole cell currents in COS-7 cells transfected with xClC-5 cDNA had strong outward rectification, Cl−> I−anion sensitivity, and were inhibited at low pH, similar to previous results in oocytes. In oocytes, antisense xClC-5 cRNA injection had no effect on endogenous membrane currents or the heterologous expression of human ClC-5. Activators of cAMP and protein kinase C inhibitors had no significant effects on ClC-5 currents expressed in either COS-7 cells or oocytes, whereas H-89, a cAMP-dependent protein kinase (PKA) inhibitor, and hydrogen peroxide decreased the currents. We conclude that the basic properties of ClC-5 currents were independent of the host cell type used for expression. In addition, ClC-5 channels may be modulated by PKA and reactive oxygen species.