Instituto de Investigaciones Bioquímicas de La Plata (INIBIOLP), Universidad Nacional de La Plata (UNLP) - CONICET CCT-La Plata, La Plata, Argentina.
Comp Biochem Physiol C Toxicol Pharmacol. 2013 May;157(4):390-6. doi: 10.1016/j.cbpc.2013.03.006. Epub 2013 Apr 6.
Hydrocarbon pollution is a major environmental threat to ecosystems in marine and freshwater environments, but its toxicological effect on aquatic organisms remains little studied. A proteomic approach was used to analyze the effect of a freshwater oil spill on the prawn Macrobrachium borellii. To this aim, proteins were extracted from midgut gland (hepatopancreas) of male and female prawns exposed 7 days to a sublethal concentration (0.6 ppm) of water-soluble fraction of crude oil (WSF). Exposure to WSF induced responses at the protein expression level. Two-dimensional gel electrophoresis (2-DE) revealed 10 protein spots that were differentially expressed by WSF exposure. Seven proteins were identified using MS/MS and de novo sequencing. Nm23 oncoprotein, arginine methyltransferase, fatty aldehyde dehydrogenase and glutathione S-transferase were down-regulated, whereas two glyceraldehyde-3-phosphate dehydrogenase isoforms and a lipocalin-like crustacyanin (CTC) were up-regulated after WSF exposure. CTC mRNA levels were further analyzed by quantitative real-time PCR showing an increased expression after WSF exposure. The proteins identified are involved in carbohydrate and amino acid metabolism, detoxification, transport of hydrophobic molecules and cellular homeostasis among others. These results provide evidence for better understanding the toxic mechanisms of hydrocarbons. Moreover, some of these differentially expressed proteins would be employed as potential novel biomarkers.
烃类污染是海洋和淡水环境中生态系统的主要环境威胁,但它对水生生物的毒性作用仍研究甚少。本研究采用蛋白质组学方法分析了淡水溢油对罗氏沼虾(Macrobrachium borellii)的影响。为此,从雄性和雌性罗氏沼虾的中肠腺(肝胰腺)中提取蛋白质,这些虾在亚致死浓度(0.6 ppm)的原油水溶性馏分(WSF)中暴露 7 天。WSF 暴露诱导了蛋白质表达水平的反应。二维凝胶电泳(2-DE)显示 10 个蛋白斑点因 WSF 暴露而差异表达。使用 MS/MS 和从头测序鉴定了 7 种蛋白质。Nm23 癌蛋白、精氨酸甲基转移酶、脂肪醛脱氢酶和谷胱甘肽 S-转移酶下调,而两种甘油醛-3-磷酸脱氢酶同工酶和脂钙蛋白样甲壳质(CTC)上调。进一步通过定量实时 PCR 分析 CTC mRNA 水平,显示 WSF 暴露后表达增加。鉴定的蛋白质参与碳水化合物和氨基酸代谢、解毒、疏水分子的转运和细胞内稳态等。这些结果为更好地理解烃类的毒性机制提供了证据。此外,其中一些差异表达的蛋白质可作为潜在的新型生物标志物。