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暴露于毒死蜱的斑马鱼(Danio rerio)胚胎中差异表达蛋白质的分析。

Analysis of differentially expressed proteins in zebrafish (Danio rerio) embryos exposed to chlorpyrifos.

作者信息

Liu Lili, Xu Yongxue, Xu Lili, Wang Jian, Wu Wei, Xu Lei, Yan Yanchun

机构信息

Graduate School of Chinese Academy of Agricultural Sciences, Beijing 100081, China.

出版信息

Comp Biochem Physiol C Toxicol Pharmacol. 2015 Jan;167:183-9. doi: 10.1016/j.cbpc.2014.10.006.

Abstract

In this study, the protein expression profiles of zebrafish embryos under chlorpyrifos (CPF) stress were investigated. Zebrafish embryos were exposed to 0.25 mg/L CPF, and embryo samples were collected until 24 h post-fertilization (hpf). To gain a better understanding of the response of zebrafish embryos to CPF exposure, two-dimensional polyacrylamide gel electrophoresis (2D PAGE) coupled with mass spectrometry was employed to carry out a comparative proteomic analysis. Total proteins were extracted from the control and treated samples, separated by 2D PAGE, and visualized by silver staining. A total of 59 protein spots showed reproducible changes compared with the control. Of these 59 spots, 19 were selected and subjected to matrix-assisted laser desorption/ionization (MALDI) tandem time-of-flight mass spectrometry (TOF/TOF) analysis; 9 differentially expressed proteins were successfully identified, including 3 up-regulated proteins and 6 down-regulated proteins. The increased expression of 3 proteins associated with detoxification and stress response suggested that the activation of protective proteins was required in zebrafish embryos exposed to CPF. On the other hand, the decreased expression of 6 proteins is mainly involved in cytoskeleton structure, protein translation, signal transduction and lipoprotein metabolism. These data may help us understand the functions and the molecular mechanisms of these proteins in zebrafish embryos' response to CPF exposure.

摘要

在本研究中,对毒死蜱(CPF)胁迫下斑马鱼胚胎的蛋白质表达谱进行了研究。将斑马鱼胚胎暴露于0.25 mg/L的CPF中,并在受精后24小时(hpf)前收集胚胎样本。为了更好地了解斑马鱼胚胎对CPF暴露的反应,采用二维聚丙烯酰胺凝胶电泳(2D PAGE)结合质谱法进行比较蛋白质组学分析。从对照样本和处理样本中提取总蛋白,通过2D PAGE分离,并用银染法进行可视化。与对照相比,共有59个蛋白点显示出可重复的变化。在这59个点中,选择了19个进行基质辅助激光解吸/电离(MALDI)串联飞行时间质谱(TOF/TOF)分析;成功鉴定出9种差异表达蛋白,包括3种上调蛋白和6种下调蛋白。3种与解毒和应激反应相关的蛋白表达增加,表明暴露于CPF的斑马鱼胚胎需要激活保护蛋白。另一方面,6种蛋白表达下降主要涉及细胞骨架结构、蛋白质翻译、信号转导和脂蛋白代谢。这些数据可能有助于我们了解这些蛋白在斑马鱼胚胎对CPF暴露反应中的功能和分子机制。

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