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极化上皮细胞系MDCK中一种类震荡器(A型)钾通道的极化靶向作用

Polarized targeting of a shaker-like (A-type) K(+)-channel in the polarized epithelial cell line MDCK.

作者信息

Le Maout S, Sewing S, Coudrier E, Elalouf J M, Pongs O, Merot J

机构信息

Department de Biologie Cellulaire et Moléculaire, CEA Saclay, Gif sur Yvette, France.

出版信息

Mol Membr Biol. 1996 Jul-Sep;13(3):143-7. doi: 10.3109/09687689609160590.

Abstract

Functional Kv 1-4 channels were stably expressed in filter-grown MDCK cells which form a polarized epithelium with two distinct plasma membrane domains: a basolateral and an apical cell surface. The Shaker-related Kv 1-4 channels mediated in MDCK cells fast transient (A-type) voltage-activated outward currents having similar properties to the ones reported for Kv 1-4 in the Xenopus oocytes expression system. Immunoblot analysis with specific anti-Kv 1-4 antibodies showed that two Kv 1-4 protein forms are expressed in MDCK cells which most likely represent the glycosylated and non-glycosylated Kv 1-4 protein, respectively. Using immunocytochemistry and confocal microscopy we showed that the Kv 1-4 channels are specifically localized in the basolateral membranes of MDCK cells. Thus, the MDCK cells may provide an important model system to analyse the polarized transport of ion channels such as Kv 1-4, which are distinctly expressed in the mammalian central nervous system.

摘要

功能性Kv 1-4通道在滤膜生长的MDCK细胞中稳定表达,MDCK细胞形成具有两个不同质膜结构域的极化上皮:基底外侧和顶端细胞表面。与Shaker相关的Kv 1-4通道在MDCK细胞中介导快速瞬时(A 型)电压激活外向电流,其性质与非洲爪蟾卵母细胞表达系统中报道的Kv 1-4电流相似。用特异性抗Kv 1-4抗体进行免疫印迹分析表明,MDCK细胞中表达两种Kv 1-4蛋白形式,它们最有可能分别代表糖基化和非糖基化的Kv 1-4蛋白。利用免疫细胞化学和共聚焦显微镜,我们发现Kv 1-4通道特异性定位于MDCK细胞的基底外侧膜。因此,MDCK细胞可能为分析离子通道(如Kv 1-4)的极化转运提供一个重要的模型系统,Kv 1-4在哺乳动物中枢神经系统中明显表达。

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