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深入研究离子通道和转运蛋白功能。秀丽隐杆线虫ClC型氯离子通道:新型变体与功能表达。

Into ion channel and transporter function. Caenorhabditis elegans ClC-type chloride channels: novel variants and functional expression.

作者信息

Nehrke K, Begenisich T, Pilato J, Melvin J E

机构信息

Center for Oral Biology, Aab Institute of Biomedical Sciences, University of Rochester Medical Center, Rochester, New York 14642, USA.

出版信息

Am J Physiol Cell Physiol. 2000 Dec;279(6):C2052-66. doi: 10.1152/ajpcell.2000.279.6.C2052.

Abstract

Six ClC-type chloride channel genes have been identified in Caenorhabditis elegans, termed clh-1 through clh-6. cDNA sequences from these genes suggest that clh-2, clh-3, and clh-4 may code for multiple channel variants, bringing the total to at least nine channel types in this nematode. Promoter-driven green fluorescent protein (GFP) expression in transgenic animals indicates that the protein CLH-5 is expressed ubiquitously, CLH-6 is expressed mainly in nonneuronal cells, and the remaining isoforms vary from those restricted to a single cell to those expressed in over a dozen cells of the nematode. In an Sf9 cell expression system, recombinant CLH-2b, CLH-4b, and CLH-5 did not form functional plasma membrane channels. In contrast, both CLH-1 and CLH-3b produced strong, inward-rectifying chloride currents similar to those arising from mammalian ClC2, but which operate over different voltage ranges. Our demonstration of multiple CLH protein variants and comparison of expression patterns among the clh gene family provides a framework, in combination with the electrical properties of the recombinant channels, to further examine the physiology and cell-specific role each isoform plays in this simple model system.

摘要

秀丽隐杆线虫中已鉴定出六个ClC型氯离子通道基因,命名为clh-1至clh-6。这些基因的cDNA序列表明,clh-2、clh-3和clh-4可能编码多种通道变体,使这种线虫中的通道类型总数至少达到九种。启动子驱动的绿色荧光蛋白(GFP)在转基因动物中的表达表明,蛋白CLH-5在全身表达,CLH-6主要在非神经细胞中表达,其余异构体的表达范围从局限于单个细胞到线虫中十几个细胞中表达不等。在Sf9细胞表达系统中,重组CLH-2b、CLH-4b和CLH-5未形成功能性质膜通道。相比之下,CLH-1和CLH-3b均产生了强大的内向整流氯离子电流,类似于哺乳动物ClC2产生的电流,但作用于不同的电压范围。我们对多种CLH蛋白变体的证明以及clh基因家族中表达模式的比较,结合重组通道的电特性,为进一步研究每种异构体在这个简单模型系统中的生理学和细胞特异性作用提供了一个框架。

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