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鉴定并检测鲫鱼(Gobio gobio)在多氯联苯 77 暴露下的 pi 类谷胱甘肽 S-转移酶和硒依赖性谷胱甘肽过氧化物酶的 mRNA 表达。

Identification and mRNA expression of pi-class glutathione S-transferase and selenium-dependent glutathione peroxidase in the gudgeon Gobio gobio exposed to PCB 77.

机构信息

Department, Environment and Agro-biotechnologies, Centre de Recherche Public-Gabriel Lippmann, rue du Brill, 4422 Belvaux, Luxembourg.

出版信息

Comp Biochem Physiol C Toxicol Pharmacol. 2012 Mar;155(2):300-6. doi: 10.1016/j.cbpc.2011.09.009. Epub 2011 Oct 1.

Abstract

In aquatic environments some pollutants are present in water and sediments and organisms possess cellular detoxification systems to face up these xenobiotics. The gudgeon, Gobio gobio, is a freshwater benthopelagic fish that appears particularly adequate for an ecotoxicological assessment of rivers. The aim of this study was the identification of GST and GPx genes in this organism in order to develop new indicators of early exposure to xenobiotics in aquatic environments. Reverse-transcription PCR using degenerate primers and RACE-PCR allowed us to identify a selenium-dependent glutathione peroxidase (Se-GPx), belonging to the class one (GPx-1), and a pi-class glutathione S-transferase (pi-GST) cDNAs. These sequences encoded for 191 and 208 amino acids proteins respectively, they exhibit high identities and similarities with corresponding proteins in other fish and the residues essential to the enzymatic functions are highly conserved. The expression patterns of these two mRNA were established by real-time PCR in five gudgeon tissues: liver, gills, kidney, spleen and muscle. The mRNA levels of these genes were highest in the liver and their expression in the other tissues exhibited some differences. Gudgeons exposed to PCB 77 in the food exhibited an increase in pi-GST mRNA and a decrease in GPx-1 transcripts levels in the liver. However, no modification of the enzymatic activities was observed. The present study provides the first transcriptional data regarding pi-GST and GPx-1 in the gudgeon Gobio gobio.

摘要

在水生环境中,一些污染物存在于水和沉积物中,而生物体拥有细胞解毒系统来应对这些外来物质。欧鳊(Gobio gobio)是一种淡水底栖洄游鱼类,对于河流的生态毒理学评估来说,它似乎是一种特别合适的生物。本研究的目的是在该生物中鉴定 GST 和 GPx 基因,以便开发水生环境中早期暴露于外来物质的新指标。使用简并引物和 RACE-PCR 的逆转录 PCR 使我们能够鉴定一种依赖于硒的谷胱甘肽过氧化物酶(Se-GPx),属于第一类(GPx-1),以及一种 pi 类谷胱甘肽 S-转移酶(pi-GST)cDNA。这些序列分别编码 191 和 208 个氨基酸的蛋白质,它们与其他鱼类的相应蛋白质具有高度的同一性和相似性,并且酶功能所必需的残基高度保守。通过实时 PCR 在欧鳊的五个组织(肝、鳃、肾、脾和肌肉)中建立了这两种 mRNA 的表达模式。这些基因的 mRNA 水平在肝脏中最高,其在其他组织中的表达存在一些差异。在食物中暴露于 PCB 77 的欧鳊肝脏中 pi-GST mRNA 增加,而 GPx-1 转录物水平降低。然而,未观察到酶活性的改变。本研究提供了关于欧鳊 Gobio gobio 中 pi-GST 和 GPx-1 的首次转录数据。

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