Schriever A M, Friedrich T, Pusch M, Jentsch T J
Zentrum für Molekulare Neurobiologie Hamburg (ZMNH), Hamburg University, Martinistrasse 85, D-20246 Hamburg, Germany.
J Biol Chem. 1999 Nov 26;274(48):34238-44. doi: 10.1074/jbc.274.48.34238.
The genome of the nematode Caenorhabditis elegans encodes six putative chloride channels (CeCLC-1 through CeCLC-6) that represent all three known branches of the mammalian CLC gene family. Using promoter fragments to drive the expression of the green fluorescent protein, CeCLC-2, -3, and -4 expression was studied in transgenic C. elegans. CeCLC-4 was specifically expressed in the large H-shaped excretory cell, where it was co-expressed with CeCLC-3, which is also expressed in other cells, including neurons, muscles, and epithelial cells. Also, CeCLC-2 was expressed in several cells of the nervous system, intestinal cells, and vulval muscle cells. Similar to mammalian CLC proteins, only two nematode CLC channels elicited detectable plasma membrane currents in Xenopus oocytes. CeCLC-3 currents were inwardly rectifying and were activated by positive prepulses. Its complex gating behavior can be explained by two gates, at least one of which depends on extracellular anions. In this respect it resembles some mammalian chloride channels with which it also shares a preference of chloride over iodide. C. elegans thus provides new opportunities to understand common mechanisms underlying structure and function in CLC channels and will allow for a genetic dissection of chloride channels in this simple model organism.
线虫秀丽隐杆线虫的基因组编码六种假定的氯离子通道(CeCLC-1至CeCLC-6),它们代表了哺乳动物CLC基因家族的所有三个已知分支。利用启动子片段驱动绿色荧光蛋白的表达,在转基因秀丽隐杆线虫中研究了CeCLC-2、-3和-4的表达。CeCLC-4在大的H形排泄细胞中特异性表达,在该细胞中它与CeCLC-3共表达,CeCLC-3也在包括神经元、肌肉和上皮细胞在内的其他细胞中表达。此外,CeCLC-2在神经系统的几个细胞、肠细胞和外阴肌细胞中表达。与哺乳动物CLC蛋白相似,只有两个线虫CLC通道在非洲爪蟾卵母细胞中引发了可检测的质膜电流。CeCLC-3电流呈内向整流,由正向预脉冲激活。其复杂的门控行为可以由两个门来解释,其中至少一个门依赖于细胞外阴离子。在这方面,它类似于一些哺乳动物氯离子通道,它也与这些通道一样更倾向于氯离子而非碘离子。因此,秀丽隐杆线虫为理解CLC通道结构和功能的共同机制提供了新的机会,并将允许在这种简单的模式生物中对氯离子通道进行遗传学剖析。