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绿白鲟幼鱼暴露于热应激和硒环境下的蛋白质组学分析。

A proteomic analysis of green and white sturgeon larvae exposed to heat stress and selenium.

机构信息

Department of Animal Science, University of California, Davis, CA 95616, USA.

出版信息

Sci Total Environ. 2010 Jul 15;408(16):3176-88. doi: 10.1016/j.scitotenv.2010.04.005.

Abstract

Temperature and selenium are two environmental parameters that potentially affect reproduction and stock recruitment of sturgeon in the San Francisco Bay/Delta Estuary. To identify proteins whose expression is modified by these environmental stressors, we performed a proteomic analysis on larval green and white sturgeons exposed to 18 or 26 degrees C and micro-injected with Seleno-L-Methionine to reach 8microgg(-)(1) selenium body burden, with L-Methionine as a control. Selenium and high temperature induced mortalities and abnormal morphologies in both species, with a higher mortality in green sturgeon. Larval proteins were separated by two-dimensional gel electrophoresis and differential abundances were detected following spot quantitation and hierarchical cluster analysis. In green sturgeon, 34 of 551 protein spots detected on gels showed a variation in abundance whereas in white sturgeon only 9 of 580 protein spots were differentially expressed (P<0.01). Gel replicates were first grouped according to heat treatment. Fifteen of these spots were identified using MALDI TOF/TOF mass spectrometry. Proteins involved in protein folding, protein synthesis, protein degradation, ATP supply and structural proteins changed in abundance in response to heat and/or selenium. 40S ribosomal protein SA, FK506-binding protein 10, 65kDa regulatory subunit A of protein phosphatase 2, protein disulfide isomerase, stress-induced-phosphoprotein 1, suppression of tumorigenicity 13 and collagen type II alpha 1, were differentially expressed in high temperature treatment only. Serine/arginine repetitive matrix protein 1, creatine kinase, serine peptidase inhibitor Kazal type 5 and HSP90 were sensitive to combined temperature and selenium exposure. Valosin-containing protein, a protein involved in aggresome formation and in protein quality control decreased more than 50% in response to selenium treatment. Potential use of such proteins as biomarkers of environmental stressors in larval sturgeons could indicate early warning signals preceding population decline.

摘要

温度和硒是两个潜在影响旧金山湾/三角洲河口鲟鱼繁殖和种群补充的环境参数。为了鉴定受这些环境胁迫影响而表达发生改变的蛋白质,我们对暴露于 18 或 26°C 并经微注射硒代蛋氨酸(达到 8μg/g 硒体负荷)或蛋氨酸(对照)的幼龄绿和白鲟进行了蛋白质组学分析。硒和高温诱导两种鱼类的死亡率和异常形态,绿鲟的死亡率更高。幼虫蛋白质用二维凝胶电泳分离,斑点定量和层次聚类分析后检测差异丰度。在绿鲟中,凝胶上检测到的 551 个蛋白质斑点中有 34 个丰度发生变化,而在白鲟中,只有 580 个蛋白质斑点中有 9 个差异表达(P<0.01)。凝胶重复首先根据热处理进行分组。使用 MALDI TOF/TOF 质谱对其中的 15 个斑点进行了鉴定。参与蛋白质折叠、蛋白质合成、蛋白质降解、ATP 供应和结构蛋白的蛋白质丰度因热和/或硒而发生变化。40S 核糖体蛋白 SA、FK506 结合蛋白 10、蛋白磷酸酶 2 的 65kDa 调节亚基 A、蛋白二硫键异构酶、应激诱导磷酸蛋白 1、肿瘤抑制因子 13 和胶原类型 II alpha 1,仅在高温处理中差异表达。丝氨酸/精氨酸重复基质蛋白 1、肌酸激酶、丝氨酸肽酶抑制剂 Kazal 类型 5 和 HSP90 对温度和硒的联合暴露敏感。参与聚集体形成和蛋白质质量控制的富含缬氨酸蛋白(valosin-containing protein),对硒处理的响应降低了 50%以上。这些蛋白质作为幼鲟环境胁迫生物标志物的潜在用途可能表明种群减少之前的早期预警信号。

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