Expression of nicotinic acetylcholine receptor subunits from parasitic nematodes in Caenorhabditis elegans.
Expression of nicotinic acetylcholine receptor subunits from parasitic nematodes in Caenorhabditis elegans.
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DOI：
10.1016/j.molbiopara.2015.12.006
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发表时间：
2015-11
影响因子：
1.5
通讯作者：
Wolstenholme AJ
中科院分区：
文献类型：
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作者：
Sloan MA;Reaves BJ;Maclean MJ;Storey BE;Wolstenholme AJ
线虫神经肌肉接头处存在的左旋咪唑敏感烟碱乙酰胆碱受体由多个不同的亚基组成，其确切组成因物种而异。我们测试了来自捻转血矛线虫和猪蛔虫的两个高度保守的烟碱受体亚基UNC-38和UNC-29拯救具有unc-38和unc-29基因零缺失突变的秀丽隐杆线虫菌株的左旋咪唑抗性和运动缺陷的能力。将寄生虫 cDNA 克隆到相关秀丽隐杆线虫启动子的下游，并通过基因枪转化引入突变菌株中。 H. contortus 的 UNC-38 亚基能够完全挽救无效缺失突变体 VC2937 (ok2896) 的运动缺陷和左旋咪唑抗性，但无法检测到表达猪曲霉 UNC-38 的线虫。 H. contortus UNC-29.1 亚基部分挽救了 unc-29 VC1944 (ok2450) 中秀丽隐杆线虫无效突变的左旋咪唑抗性，但在震荡测定中确实导致运动性增加。相比之下，只有含有猪曲霉UNC-29亚基的单一线虫显示出对左旋咪唑抗性的部分挽救，但对鞭打没有影响。
The levamisole-sensitive nicotinic acetylcholine receptor present at nematode neuromuscular junctions is composed of multiple different subunits, with the exact composition varying between species. We tested the ability of two well-conserved nicotinic receptor subunits, UNC-38 and UNC-29, from Haemonchus contortus and Ascaris suum to rescue the levamisole-resistance and locomotion defects of Caenorhabditis elegans strains with null deletion mutations in the unc-38 and unc-29 genes. The parasite cDNAs were cloned downstream of the relevant C. elegans promoters and introduced into the mutant strains via biolistic transformation. The UNC-38 subunit of H. contortus was able to completely rescue both the locomotion defects and levamisole resistance of the null deletion mutant VC2937 (ok2896), but no C. elegans expressing the A. suum UNC-38 could be detected. The H. contortus UNC-29.1 subunit partially rescued the levamisole resistance of a C. elegans null mutation in unc-29 VC1944 (ok2450), but did cause increased motility in a thrashing assay. In contrast, only a single line of worms containing the A. suum UNC-29 subunit showed a partial rescue of levamisole resistance, with no effect on thrashing.