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首页> 外文期刊>Australasian journal of ecotoxicology: the official journal of the Australasian Society for Ecotoxicology >PRELIMINARY ASSESSMENT OF AN INDIRECT ELISA AND WESTERN BLOT TECHNIQUES TO SCREEN VITELLOGENIN IN BARRAMUNDI AS A POTENTIAL BIOMARKER OF EXPOSURE TO EDCS IN AUSTRALIAN TROPICAL WATERS
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PRELIMINARY ASSESSMENT OF AN INDIRECT ELISA AND WESTERN BLOT TECHNIQUES TO SCREEN VITELLOGENIN IN BARRAMUNDI AS A POTENTIAL BIOMARKER OF EXPOSURE TO EDCS IN AUSTRALIAN TROPICAL WATERS

机译：间接酶联免疫吸附测定和西方印迹技术对巴拉墨地中的玻璃蛋白进行筛查的初步评估，作为在澳大利亚热带水域暴露于EDCS的潜在生物标志物

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Vitellogenin (Vtg) is a complex phospholipoglycoprotein that is involved in yolk synthesis in oviparous vertebrates. Vtg is secreted from the liver of female fish in response to ovarian estradiol and males exposed to estrogenic compounds can also produce Vtg. Endocrine disrupting chemicals (EDCs) have been identified in coastal waters of Australia, necessitating the evaluation of biomarkers, such as Vtg, for exposure of Australian finfish to EDCs. Initially, Vtg induction was assessed in plasma from 17beta-estradiol (E_2)-dosed hatchery-reared juvenile barramundi (Lates calcarifer) by indirect ELISA using a monoclonal antibody (Mab) which cross reacts to barramundi Vtg (ND-3G2). E_2-dosed juvenile barramundi (20mg/kg) demonstrated significantly higher Vtg induction compared with corn-oil injected controls (p < 0.001). These results were confirmed by Western blot. Field assessment of Vtg induction in barramundi from four rivers in Northern Queensland (Johnstone, Herbert, Lockhart and Pascoe) demonstrated higher Vtg levels than laboratory control fish, albeit not significant. Juvenile_male_barramundi_from_two sites in Darwin Harbour (Sadgroves Creek and West Arm) had Vtg levels comparable to juvenile fish sampled from a local hatchery and juvenile laboratory control fish (p > 0.05). This work represents one of the first evaluations of ELISA and Western blot techniques for assessment of Vtg induction in barramundi and demonstrates the potential use of Vtg induction as a biomarker for exposure to EDCs in Australian coastal waters. With further development of Australian species-specific antibodies and Vtg standards, this assay could be a very powerful and sensitive diagnostic tool for Australian waters.
机译：卵黄蛋白原（Vtg）是一种复杂的磷酸脂糖蛋白，与卵生脊椎动物的卵黄合成有关。 Vtg是雌性鱼类肝脏中对卵巢雌二醇的反应所分泌的，而雄性接触雌激素化合物也可以产生Vtg。已在澳大利亚沿海水域发现了破坏内分泌的化学物质（EDC），因此有必要评估诸如Vtg的生物标志物，以使澳大利亚有鳍鱼类接触EDC。最初，通过间接ELISA使用单克隆抗体（Mab）与bar鱼Vtg（ND-3G2）发生交叉反应，通过间接ELISA对血浆中17β-雌二醇（E_2）饲养的孵化的幼鱼澳洲肺鱼（Lates calcarifer）的Vtg诱导进行了评估。与玉米油注射的对照组相比，E_2剂量的幼鱼（20mg / kg）表现出明显更高的Vtg诱导（p <0.001）。这些结果通过蛋白质印迹证实。昆士兰北部昆士兰州的四条河流（约翰斯通，赫伯特，洛克哈特和帕斯科地区）的澳洲肺鱼中Vtg诱导的现场评估显示，Vtg水平高于实验室控制鱼，尽管水平不高。在达尔文港的两个地点（Sadgroves Creek和West Arm）的Juvenile_male_barramundi_from_from地点的Vtg水平可与从当地孵化场和少年实验室控制鱼中取样的幼鱼相媲美（p> 0.05）。这项工作代表了首次评估ELISA和蛋白质印迹技术以评估澳洲肺鱼中的Vtg诱导的方法之一，并证明了Vtg诱导作为暴露于澳大利亚沿海水域EDC中的生物标记物的潜在用途。随着澳大利亚物种特异性抗体和Vtg标准的进一步发展，该测定法可能成为澳大利亚水域非常强大且灵敏的诊断工具。

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