Clone, identification and functional characterization of a novel toll (Shtoll1) from the freshwater crab Sinopotamon henanense in response to cadmium exposure and Aeromonas hydrophila infection.
Clone, identification and functional characterization of a novel toll (Shtoll1) from the freshwater crab Sinopotamon henanense in response to cadmium exposure and Aeromonas hydrophila infection.
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DOI：
10.1016/j.fsi.2020.01.012
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发表时间：
2020-01
影响因子：
4.7
通讯作者：
Lang Lang-Lang;Minnan Bao;Wei-xin Jing;Wei Chen;Lan Wang
中科院分区：
文献类型：
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作者：
Lang Lang-Lang;Minnan Bao;Wei-xin Jing;Wei Chen;Lan Wang
Toll 在先天免疫系统中至关重要，对于无脊椎动物抵御细菌、真菌和病毒感染非常重要。我们之前的研究表明，镉（Cd）可以改变嗜水气单胞菌感染的上皮（淡水蟹河南中华蟹的鳃和中肠）中ShToll3的表达模式。为了研究甲壳类同源物 Tolls 的多种先天免疫作用，在本研究中，我们从 S 中克隆了 Shtoll1。河南。 Shtoll1的cDNA全长为4746 bp，ORF为3033 bp，编码111个氨基酸的推定蛋白、255 bp的5'非翻译区和1713 bp的3'非翻译区。系统进化分析显示ShToll1聚类到DmToll1、DmToll 4和DmToll 5组中。此外，组织分布结果显示Shtoll1在不同组织中广泛表达，其中在血细胞中表达量最高。此外，A.A.后血细胞和肝胰腺中Shtoll1表达上调。亲水感染。同时，检测了血细胞中Shtoll1表达的增加，以响应Cd暴露和A。联合亲水感染。通过蛋白质印迹和免疫组织化学分析，血细胞中的ShToll1表达因A.亲水性和Cd独立地以及组合地。此外，还检测了可能对Cd和A有反应的三种抗菌肽(AMP)alf5、alf6和c-lys的mRNA水平。分析了Shtoll1对亲水刺激的影响。由此，我们得出结论，Cd扩大了ShToll1对A的敏感性。血细胞中的亲水感染。这表明ShToll1可能有助于S的先天免疫防御。河南队对抗A。血细胞中的亲水性和镉。
Toll is essential in innate immune system which is important for defense against bacterial, fungal and viral infections in invertebrates. Our previous study showed that cadmium (Cd) could change the expression pattern of ShToll3 in the epithelium (gills and midgut from the freshwater crabSinopotamon henanense) infected by Aeromonas hydrophila. To investigate the diverse innate immune roles of crustacean homolog Tolls, in this study, we clonedShtoll1fromS. henanense. The full-length cDNA ofShtoll1was 4746 bp, with an ORF of 3033 bp encoding a putative protein of 111 amino acids, a 5′-untranslated region of 255 bp and a 3′-untranslated region of 1713 bp. Phylogenetic analysis showed that ShToll1 was clustered into the group of DmToll1, DmToll 4 and DmToll 5. In addition, the tissue distribution results showed thatShtoll1was expressed widely in different tissues, with the highest expression in heamocytes. Besides,Shtoll1expressions were upregulated in heamocytes and hepatopancreas afterA. hydrophilainfection. At the same time, the increase ofShtoll1expressions were examined in heamocytes in response to Cd exposure andA. hydrophilainfection in combination. Through western blotting and immunohistochemical analysis, the ShToll1 expressions in heamocytes were increased in response toA. hydrophilaand Cd independently as well as in combination. Moreover, the mRNA level of three antimicrobial peptides (AMPs)alf5, alf6,andc-lys, which possibly responded to Cd andA. hydrophilastimulation throughShtoll1,were analyzed.Thus, we conclude that Cd expand the susceptibility of ShToll1 toA. hydrophilainfection in heamocytes. This suggest that ShToll1 may contribute to the innate immune defense ofS. henanenseagainstA. hydrophilaand Cd in heamocytes.