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大黄鱼金属硫蛋白:在睾丸中的表达及对镉诱导损伤的反应

Metallothionein from Pseudosciaena crocea: expression and response to cadmium-induced injury in the testes.

作者信息

Sheng Zhang, Yang Wan-Xi, Zhu Jun-Quan

机构信息

School of Marine Sciences, Ningbo University, Ningbo, 315211, Zhejiang, People's Republic of China.

出版信息

Ecotoxicology. 2015 May;24(4):779-94. doi: 10.1007/s10646-015-1423-y. Epub 2015 Feb 14.

Abstract

Metallothioneins (MTs) are a family of stress proteins that are involved in the process of detoxification and anti-oxidation. Previous studies have focused mostly on the expression and functions of MTs in the non-reproductive tissues of aquatic vertebrates. However, there have been only a few reports regarding the functions of MTs in the reproductive tissues of such vertebrates. In order to investigate the function of MTs during spermatogenesis in Pseudosciaena crocea, reverse-transcription polymerase chain reaction (PCR) and rapid amplification of cDNA ends were performed to obtain the P. crocea MT complete cDNA sequence from the total RNA of the testes for the first time. MT was detected in the liver, kidneys, testes, spleen, gill and muscle of P. crocea by tissue-specific expression analysis. Meanwhile, immunohistochemistry staining indicated that the MT protein was localized in germ cells, Sertoli cells and the peripheral connective tissues in P. crocea testes. Furthermore, acute toxicity tests were conducted with cadmium (Cd) to determine the 96 h-medial lethal concentration value. The toxic effects of Cd on the microstructure and ultrastructure of the testes were observed. In addition, the changes in MT mRNA expression levels in the testes after Cd exposure were measured using real-time quantitative PCR. Consequently, we suggest that MTs play an important role in spermatogenesis and testes protection against Cd toxicity in P. crocea.

摘要

金属硫蛋白(MTs)是一类应激蛋白,参与解毒和抗氧化过程。以往的研究主要集中在MTs在水生脊椎动物非生殖组织中的表达和功能。然而,关于MTs在这类脊椎动物生殖组织中的功能报道较少。为了研究大黄鱼精子发生过程中MTs的功能,首次通过逆转录聚合酶链反应(PCR)和cDNA末端快速扩增从睾丸总RNA中获得大黄鱼MT完整cDNA序列。通过组织特异性表达分析在大黄鱼的肝脏、肾脏、睾丸、脾脏、鳃和肌肉中检测到MT。同时,免疫组织化学染色表明MT蛋白定位于大黄鱼睾丸的生殖细胞、支持细胞和外周结缔组织中。此外,用镉(Cd)进行急性毒性试验以确定96小时半数致死浓度值。观察了Cd对睾丸微观结构和超微结构的毒性作用。另外,使用实时定量PCR测量Cd暴露后睾丸中MT mRNA表达水平的变化。因此,我们认为MTs在大黄鱼精子发生和睾丸抵御Cd毒性中起重要作用。

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