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Proteomic Analysis of the Reproductive Organs of the Hermaphroditic Gastropod Lymnaea stagnalis Exposed to Different Endocrine Disrupting Chemicals

机译:暴露于不同内分泌干扰物的雌雄同体腹足动物生殖器生殖器官的蛋白质组学分析

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摘要

Many studies have reported perturbations of mollusc reproduction following exposure to low concentrations (ng/L range) of endocrine disrupting chemicals (EDCs). However, the mechanisms of action of these molecules on molluscs are still poorly understood. Investigation of the modifications of protein expression in organisms exposed to chemicals using proteomic methods can provide a broader and more comprehensive understanding of adverse impacts of pollution on organisms than conventional biochemical biomarkers (e.g., heat-shock proteins, metallothioneins, GST, EROD). In this study we have investigated the impacts of four chemicals, which exhibit different endocrine disrupting properties in vertebrates, on the proteome of the hermaphroditic freshwater pulmonate gastropod Lymnaea stagnalis after 21 days of exposure. Testosterone, tributyltin, chlordecone and cyproterone acetate were chosen as tested compounds as they can induce adverse effects on the reproduction of this snail. The 2D-DIGE method was used to identify proteins whose expression was affected by these compounds. In addition to modifying the expression of proteins involved in the structure and function of the cytoskeleton, chemicals had impacts on the expression of proteins involved in the reproduction of L. stagnalis. Exposure to 19.2 µg/L of chlordecone increased the abundance of ovipostatin, a peptide transmitted during mating through seminal fluid, which reduces oviposition in this species. The expression of yolk ferritin, the vitellogenin equivalent in L. stagnalis, was reduced after exposure to 94.2 ng Sn/L of tributyltin. The identification of yolk ferritin and the modification of its expression in snails exposed to chemicals were refined using western blot analysis. Our results showed that the tested compounds influenced the abundance of yolk ferritin in the reproductive organs. Alteration in proteins involved in reproductive pathways (e.g., ovipostatin and yolk ferritin) could constitute relevant evidence of interaction of EDCs with reproductive pathways that are under the control of the endocrine system of L. stagnalis.
机译:许多研究报告称,暴露于低浓度(ng / L范围)的内分泌干扰物(EDC)后,软体动物繁殖受到干扰。但是,这些分子对软体动物的作用机理仍知之甚少。与常规生化生物标记物(例如热休克蛋白,金属硫蛋白,GST,EROD)相比,使用蛋白质组学方法研究暴露于化学物质的生物中蛋白质表达的修饰可以提供更广泛和更全面的了解污染对生物的不利影响。在这项研究中,我们研究了暴露于21天后四种在脊椎动物中表现出不同内分泌干扰特性的化学物质对雌雄同体淡水肺腹足纲腹足Ly的蛋白质组的影响。选择睾丸激素,三丁锡,十氯酮和醋酸环丙孕酮作为被测化合物,因为它们会对这种蜗牛的繁殖产生不利影响。 2D-DIGE方法用于鉴定表达受这些化合物影响的蛋白质。除了修饰与细胞骨架的结构和功能有关的蛋白质的表达外,化学物质还对与stagnalis繁殖有关的蛋白质的表达产生影响。暴露于19.2 µg / L的十氯酮会增加ovipostatin的含量,ovipostatin是一种在交配过程中通过精液传播的肽,可减少该物种的产卵。暴露于94.2 ng Sn / L的三丁基锡后,卵黄铁蛋白的表达降低,而卵黄铁蛋白的表达等同于stagnalis中的卵黄蛋白原。蛋黄铁蛋白的鉴定及其在暴露于化学物质中的蜗牛中表达的修饰使用蛋白质印迹分析进行了完善。我们的结果表明,受试化合物影响生殖器官中卵黄铁蛋白的丰度。生殖途径中涉及的蛋白质(例如,卵黄质和卵黄铁蛋白)的改变可能构成EDC与受stagnalis内分泌系统控制的生殖途径相互作用的相关证据。

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