Identification of a novel toll gene (Shtoll3) from the freshwater crab Sinopotamon henanense and its expression pattern changes in response to cadmium followed by Aeromonas hydrophila infection
Identification of a novel toll gene (Shtoll3) from the freshwater crab Sinopotamon henanense and its expression pattern changes in response to cadmium followed by Aeromonas hydrophila infection
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DOI：
10.1016/j.fsi.2017.10.013
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发表时间：
2017-12
影响因子：
4.7
通讯作者：
Lang Lang-Lang;Zuobing Zhang;Wei-xin Jing;Jiang‐Shiou Hwang;Shao-Chin Lee;Lan Wang
中科院分区：
文献类型：
--
作者：
Lang Lang-Lang;Zuobing Zhang;Wei-xin Jing;Jiang‐Shiou Hwang;Shao-Chin Lee;Lan Wang
Toll 信号对于免疫基因的表达至关重要，而免疫基因对于无脊椎动物抵御细菌、真菌和病毒感染非常重要。尽管在甲壳类动物中已鉴定出多种toll基因，但尚未在淡水蟹河南中华蟹中研究过其中任何一个。此外，镉（Cd）ontoll基因表达对生活在沉积物中且易于重金属生物富集的淡水蟹的影响尚未得到研究。我们对肝胰腺组织的转录组分析表明，用镉处理后，toll3 基因表达降低。在这项研究中，我们从螃蟹中克隆了 onetoll 基因（特此指定为 Shtoll3）。 Shtoll3的cDNA全长为4488bp，ORF为3693bp，编码1230个氨基酸的推定蛋白，5'非翻译区414bp，3'非翻译区781bp。系统发育分析表明ShToll3 聚类到DmToll8 组中。组织分布结果显示，Shtoll3在不同组织中广泛表达，在鳃中表达最高，在血细胞中表达最低。嗜水气单胞菌感染后，仅中肠中Shtoll3表达下调。当 Cd 存在时，Shtoll3 表达响应 A。中肠和鳃中的亲水蛋白上调，蛋白质印迹分析进一步证实了这一点。此外，两种抗菌肽（AMP）crustinandc-lys的mRNA水平可能对Cd和A有反应。分析了Shtoll3对亲水刺激的影响。由此，我们得出结论，Cd改变了Shtoll3对A的敏感性。鳃和中肠的亲水感染。这表明Shtoll3可能有助于S的先天免疫防御。河南塞托A.亲水性和Cd可以改变上皮的免疫功能。
Toll signaling is essential for expression of immune genes which are important for defense against bacterial, fungal and viral infections in invertebrates. Although severaltollgenes have been identified in the crustaceans, none of them has been investigated in freshwater crabSinopotamon henanense.Moreover, the effect of cadmium (Cd) ontollgene expression has never been examined on the freshwater crabs which live in the sediments and are prone to heavy metal bioaccumulation. Our transcriptomic analysis of hepatopancreas tissue reveals thattoll3gene expression has been decreased when treated with Cd. In this study, we cloned onetollgene (hereby designatedShtoll3) from the crab. The full-length cDNA ofShtoll3was 4488 bp, with an ORF of 3693 bp encoding a putative protein of 1230 amino acids, a 5′-untranslated region of 414 bp and a 3′-untranslated region of 781 bp. Phylogenetic analysis showed that ShToll3 was clustered into the group of DmToll8. The tissue distribution results showed thatShtoll3was expressed widely in different tissues, with the highest in gills, and the lowest in hemocytes.Shtoll3expression was down-regulated only in midguts after Aeromonas hydrophila infection. With Cd presence,Shtoll3expression in response toA. hydrophilawere up-regulated in midguts and gills, which was further confirmed by western blotting analysis. Moreover, the mRNA level of two antimicrobial peptides (AMPs)crustinandc-lys, which possibly responded to Cd andA. hydrophilastimulation throughShtoll3,were analysised.Thus, we conclude that Cd changes the susceptibility ofShtoll3toA. hydrophilainfection in gills and midguts. This suggest thatShtoll3may contribute to the innate immune defense ofS. henanensetoA. hydrophilaand Cd can modify the immune function in epithelium.