Faculty of Science, University of Ontario Institute of Technology, Oshawa, Ontario, Canada.
J Neurochem. 2010 Jun;113(5):1113-22. doi: 10.1111/j.1471-4159.2010.06651.x. Epub 2010 Feb 23.
We have identified two genes from the parasitic nematode Haemonchus contortus, Hco-unc-49B and Hco-unc-49C that encode two GABA-gated chloride channel subunits. Electrophysiological analysis revealed that this channel has properties similar to those of the UNC-49 channel from the free-living nematode Caenorhabditis elegans. For example, the Hco-UNC-49B subunit forms a functional homomeric channel that responds to GABA and is highly sensitive to picrotoxin. Hco-UNC-49C alone does not respond to GABA but can assemble with Hco-UNC-49B to form a heteromeric channel with a lower sensitivity to picrotoxin. However, we did find that the Hco-UNC-49B/C heteromeric channel is significantly more responsive to agonists compared to the Hco-UNC-49B homomeric channel, which is the opposite trend to what has been found previously for the C. elegans channel. To investigate the subunit requirements for high agonist sensitivity, we generated cross-assembled channels by co-expressing the H. contortus subunits with UNC-49 subunits from C. elegans (Cel-UNC-49). Co-expressing Cel-UNC-49B with Hco-UNC-49C produced a heteromeric channel with a reduced sensitivity to GABA compared to that of the Cel-UNC-49B homomeric channel. In contrast, co-expressing Hco-UNC-49B with Cel-UNC-49C produced a heteromeric channel that, like the Hco-UNC-49B/C heteromeric channel, exhibits an increased sensitivity to GABA. These results suggest that the Hco-UNC-49B subunit is the key determinant for the high agonist sensitivity of heteromeric channels.
我们从寄生线虫捻转血矛线虫中鉴定出两个基因,Hco-unc-49B 和 Hco-unc-49C,它们分别编码两个 GABA 门控氯离子通道亚基。电生理分析表明,该通道具有与自由生活线虫秀丽隐杆线虫的 UNC-49 通道相似的特性。例如,Hco-UNC-49B 亚基形成一种对 GABA 有反应且对印防己毒素高度敏感的功能性同源通道。Hco-UNC-49C 本身不能对 GABA 作出反应,但可以与 Hco-UNC-49B 组装形成对印防己毒素敏感性较低的异源通道。然而,我们确实发现 Hco-UNC-49B/C 异源通道对激动剂的反应性明显高于 Hco-UNC-49B 同源通道,这与先前在秀丽隐杆线虫通道中发现的趋势相反。为了研究高激动剂敏感性的亚基要求,我们通过共表达捻转血矛线虫的亚基与秀丽隐杆线虫的 UNC-49 亚基(Cel-UNC-49)生成了交叉组装的通道。与 Cel-UNC-49B 共表达 Hco-UNC-49C 产生的异源通道对 GABA 的敏感性比 Cel-UNC-49B 同源通道降低。相比之下,与 Cel-UNC-49C 共表达 Hco-UNC-49B 产生的异源通道与 Hco-UNC-49B/C 异源通道一样,对 GABA 的敏感性增加。这些结果表明,Hco-UNC-49B 亚基是异源通道高激动剂敏感性的关键决定因素。