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Differential GFP Expression Patterns Induced by Different Heavy Metals in Tg(hsp70:gfp) Transgenic Medaka (Oryzias latipes)

机译:Tg(hsp70:gfp)转基因Medaka(Oryzias latipes)中不同重金属诱导的GFP差异表达模式

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Heat shock protein 70 (Hsp70) is one of the most widely used biomarker for monitoring environment perturbations in biological systems. To facilitate the analysis of hsp70 expression as a biomarker, we generated a Tg(hsp70:gfp) transgenic medaka line in which green fluorescence protein (GFP) reporter gene was driven by the medaka hsp70 promoter. Here, we characterized Tg(hsp70:gfp) medaka for inducible GFP expression by seven environment-relevant heavy metals, including mercury, arsenic, lead, cadmium, copper, chromium, and zinc. We found that four of them (mercury, arsenic, lead, and cadmium) induced GFP expression in multiple and different organs. In general, the liver, kidney, gut, and skin are among the most frequent organs to show induced GFP expression. In contrast, no detectable GFP induction was observed to copper, chromium, or zinc, indicating that the transgenic line was not responsive to all heavy metals. RT-qPCR determination of hsp70 mRNA showed similar induction and non-induction by these metals, which also correlated with the levels of metal uptake in medaka exposed to these metals. Our observations suggested that these heavy metals have different mechanisms of toxicity and/or differential bioaccumulation in various organs; different patterns of GFP expression induced by different metals may be used to determine or exclude metals in water samples tested. Furthermore, we also tested several non-metal toxicants such as bisphenol A, 2,3,7,8-tetrachlorodibenzo-p-dioxin, 4-introphenol, and lindane; none of them induced significant GFP expression in Tg(hsp70:gfp) medaka, further suggesting that the inducibility of Tg(hsp70:gfp) for GFP expression is specific to a subset of heavy metals.
机译:热激蛋白70(Hsp70)是用于监测生物系统中环境扰动的最广泛使用的生物标志物之一。为了便于分析作为生物标志物的hsp70表达,我们生成了Tg(hsp70:gfp)转基因medaka品系,其中绿色荧光蛋白(GFP)报告基因由medaka hsp70启动子驱动。在这里,我们将Tg(hsp70:gfp)medaka表征为可诱导的GFP表达的七种与环境相关的重金属,包括汞,砷,铅,镉,铜,铬和锌。我们发现其中的四个(汞,砷,铅和镉)在多个不同的器官中诱导了GFP表达。通常,肝脏,肾脏,肠和皮肤是显示诱导的GFP表达的最常见器官。相反,未观察到对铜,铬或锌的可检测到的GFP诱导,表明该转基因系对所有重金属均无反应。 RT-qPCR测定的hsp70 mRNA显示出这些金属具有相似的诱导和非诱导作用,这也与暴露于这些金属的青aka中的金属吸收水平相关。我们的观察结果表明,这些重金属在不同器官中具有不同的毒性机制和/或不同的生物蓄积机制。由不同金属诱导的GFP表达的不同模式可用于确定或排除测试的水样品中的金属。此外,我们还测试了几种非金属毒物,例如双酚A,2,3,7,8-四氯二苯并-对-二恶英,4-吲哚酚和林丹;它们中没有一个在Tg(hsp70:gfp)medaka中诱导显着的GFP表达,进一步表明Tg(hsp70:gfp)对GFP表达的诱导性是特定于重金属子集的。

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