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镉暴露下条纹小鲃肝脏金属硫蛋白的分子克隆、特征分析及表达分析

Molecular cloning, characterization and expression analysis of metallothionein in the liver of the teleost Acrossocheilus fasciatus exposed to cadmium chloride.

作者信息

Liu Guo-Di, Sheng Zhang, Hou Cong-Cong, Ni Jie, Han Ying-Li, Wang You-Fa, Zhou Yang, Fu Su-Yan, Zhu Jun-Quan

机构信息

School of Marine Sciences, Ningbo University, Ningbo, Zhejiang 315211, PR China.

School of Marine Sciences, Ningbo University, Ningbo, Zhejiang 315211, PR China.

出版信息

Environ Toxicol Pharmacol. 2017 Jul;53:1-9. doi: 10.1016/j.etap.2017.03.020. Epub 2017 Mar 29.

Abstract

Metallothionein (MT) has a characteristic molecular structure with a cysteine-rich content. This unique structure provides metal-binding and redox capabilities and promoting metal homeostasis and detoxification in living animals. In order to evaluate the effects of cadmium (Cd) on hepatic MT expression in the liver of Acrossocheilus fasciatus, we obtained the complete cDNA of the A. fasciatus liver MT for the first time. The MT cDNA contains a 605-bp sequence, which codes for 60 amino acids. Protein alignment showed that the similarity between MT protein sequences of A. fasciatus and those of other vertebrates (especially teleosts) was very high and a cysteine residue structure was also conserved. MT was detected in the liver, kidney, gill, testis, muscle, spleen, heart and brain tissues of A. fasciatus by tissue-specific expression analysis. After Cd exposure, Cd/hemoglobin saturation assay, immunohistochemistry and reverse-transcription quantitative PCR (RT-qPCR) were used to describe MT expression in liver tissue. These techniques indicate a sensitive response by liver MT to Cd exposure. The results suggest that A. fasciatus MT may play an important role in the detoxification processes in the liver, and also would be a useful biomarker for monitoring metal pollution in aquatic environments. In addition, A. fasciatus could be regarded as one candidate for a model species for bony fishes in ecotoxicology.

摘要

金属硫蛋白(MT)具有富含半胱氨酸的特征性分子结构。这种独特的结构赋予其金属结合和氧化还原能力,有助于维持活体动物体内的金属稳态并实现解毒作用。为了评估镉(Cd)对条纹光唇鱼肝脏中MT表达的影响,我们首次获得了条纹光唇鱼肝脏MT的完整cDNA。该MT cDNA包含一个605个碱基对的序列,编码60个氨基酸。蛋白质比对显示,条纹光唇鱼的MT蛋白序列与其他脊椎动物(尤其是硬骨鱼)的MT蛋白序列相似度很高,且半胱氨酸残基结构也保守。通过组织特异性表达分析,在条纹光唇鱼的肝脏、肾脏、鳃、睾丸、肌肉、脾脏、心脏和脑组织中检测到了MT。镉暴露后,采用镉/血红蛋白饱和度测定、免疫组织化学和逆转录定量PCR(RT-qPCR)来描述肝脏组织中MT的表达情况。这些技术表明肝脏MT对镉暴露有敏感反应。结果表明,条纹光唇鱼的MT可能在肝脏解毒过程中发挥重要作用,并且有望成为监测水生环境中金属污染的有用生物标志物。此外,条纹光唇鱼可被视为生态毒理学中硬骨鱼模型物种的候选之一。

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