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首页> 外文期刊>Aquatic Toxicology >Expression and function analysis of metallothionein in the testis of Portunus trituberculatus exposed to cadmium
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Expression and function analysis of metallothionein in the testis of Portunus trituberculatus exposed to cadmium

机译：金属硫蛋白在暴露于镉的三疣梭子蟹睾丸中的表达及功能分析

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Metallothioneins (MTs) possess a unique molecular structure that provides metal-binding and redox capabilities. These capabilities include the maintenance of metal equilibria that protect against heavy metals (especially cadmium) and oxidative damage. Past studies have focused on the function of MTs in vertebrates. However, the functions of MTs during spermiogenesis in invertebrates remain unclear. In order to investigate the function of MTs during spermiogenesis in Portunus trituberculatus, we used RTPCR and RACE to identify two MT complete cDNA sequences in the total RNA from the P. trituberculatus testis. The 450 bp MT-1 cDNA consists of a 77 bp 5' untranslated region, a 196 bp 3' untranslated region, and a 177 bp open reading frame that encodes 58 amino acids including 19 cysteines. The 581 bp MT-2 cDNA consists of 73 bp 5' untranslated region, a 328 bp 3' untranslated region, and a 180 bp open reading frame that encodes 59 amino acids including 18 cysteines. MT-1 and MT-2 of P. trituberculatus more closely resemble invertebrate (especially crab) MT homologues than vertebrate MT homologues as indicated by protein alignment comparisons and phylogenetic tree analysis. MT-1 and MT-2 were detected in the heart, testis, muscle, hepatopancreas, and gill of P. trituberculatus by tissue expression analysis. In addition, MT-1 and MT-2 are present during the entire process of spermiogenesis in P. trituberculatus as indicated by H&E staining and in situ hybridization. MT-1 and MT-2 expression levels significantly increase after cadmium (Cd) exposure as measured by real-time quantitative PCR analysis. Therefore, we suggest that MT-1 and MT-2 perform important functions in spermiogenesis and testis detoxification in P. trituberculatus. (C) 2013 Elsevier B.V. All rights reserved.
机译：金属硫蛋白（MTs）具有独特的分子结构，可提供金属结合和氧化还原功能。这些功能包括维持金属平衡，防止重金属（尤其是镉）和氧化损伤。过去的研究集中于MT在脊椎动物中的功能。但是，MTs在无脊椎动物的精子发生过程中的功能仍不清楚。为了研究三疣梭子蟹精子发生过程中MTs的功能，我们使用RTPCR和RACE鉴定了三疣梭子蟹睾丸总RNA中的两个MT完整cDNA序列。 450 bp的MT-1 cDNA由一个77 bp的5'非翻译区，一个196 bp的3'非翻译区和一个177 bp的开放阅读框组成，其编码58个氨基酸，包括19个半胱氨酸。 581 bp MT-2 cDNA由73 bp的5'非翻译区，328 bp的3'非翻译区和180 bp的开放阅读框组成，其编码59个氨基酸，包括18个半胱氨酸。如蛋白比对比较和系统进化树分析所表明的，三核对虾的MT-1和MT-2比无脊椎动物（尤其是螃蟹）的MT同源物更像脊椎动物的MT同源物。通过组织表达分析在心脏，睾丸，肌肉，肝胰脏和tri中检测到MT-1和MT-2。另外，如H＆E染色和原位杂交所示，MT-1和MT-2存在于三疣梭子蟹的整个精子发生过程中。通过实时定量PCR分析测得，镉（Cd）暴露后MT-1和MT-2表达水平显着增加。因此，我们建议MT-1和MT-2在P. trituberculatus的精子发生和睾丸排毒中起重要作用。 （C）2013 Elsevier B.V.保留所有权利。

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