环境雌激素对鱼类Kisspeptin/GnRH系统的神经毒性及作用机理研究

项目名称： 环境雌激素对鱼类Kisspeptin/GnRH系统的神经毒性及作用机理研究

项目编号： No.31470554

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 生物科学

项目作者： 杨明

作者单位： 上海大学

项目金额： 86万元

中文摘要： 双酚A和烷基酚类化合物是由人类活动引入水生生态系统的环境雌激素，已确认对水生生物特别是鱼类具有显著的内分泌干扰效应。申请人前期的研究发现该类物质对鱼类具有神经毒性，如影响胚胎神经发育相关基因的表达、诱导神经细胞凋亡、影响kisspeptin/GnRH系统基因的表达水平等。本项目拟在前期工作的基础上，以模式生物野生斑马鱼和绿色荧光标记的GnRH转基因斑马鱼为研究对象，以双酚A和壬基酚为目标物，设置环境相关浓度下单独暴露和联合暴露的在体和离体神经毒性实验，并通过拮抗剂共暴露和受体基因敲除等策略，结合多色荧光标记和共聚焦显微技术，从细胞和分子水平探索鱼类中枢神经系统在下丘脑-垂体-性腺轴上游的kisspeptin/GnRH调控系统对该类化合物的毒性响应及其作用途径和调控机理。本项目预期从根本上阐明环境雌激素类物质对鱼类内分泌干扰效应及神经毒性发生的根源，为保障鱼类种群繁育具有重要的科学意义。

中文关键词： 双酚A；烷基酚；斑马鱼；神经毒性；Kisspeptin/GnRH系统

英文摘要： Bisphenol A and alkylphenols are environmental estrogen-like compounds being introduced into the aquatic ecosystems by human activities. Those chemicals have been confirmed by numbers of previous studies to have significant endocrine disrupting effects on aquatic organisms, especially on fish. Our previous studies have shown that those chemicals are also neurotoxic substances to fish. Upon exposure to those compounds, the expression of genes related to the development of nervous system in fish embryos has been changed, and cell apoptosis has been induced, and the expression levels of kisspeptin / GnRH system-related genes have been altered. According to those results, the present project is designed to evaluate the effects of bisphenol A and nonylphenol on kisspeptin / GnRH neural system in fish under individual and joint exposure to those chemicals at environmental relevant concentrations. The in vivo and in vitro neurotoxicity experiments will be set using the lived whole mount GnRH transgenic zebrafish embryos and the primary neural cells cultured from GnRH transgenic zebrafish embryos. The combined exposure with estrogen receptor antagonist and receptor knock-out strategies will be implemented. Using multicolor fluorescence labeling and confocal microscopy imaging methods, the project aims to explore the toxic response, signaling pathway and its role in the regulation mechanism of kisspeptin / GnRH system, the upstream regulatory neural system in the central nervous system modulating the response of Hypothalamic - Pituitary - Gonadal (HPG) axis in fish. The project is expected to clarify the action mechanisms of environmental estrogen-like chemicals regarding their endocrine disrupting effects and neurotoxicity on fish. Furthermore, these results should be of great scientific significance for breeding and population conservation for fish stocks.

英文关键词： Bisphenol A;Alkylphenol;Zebrafish;Neurotoxicity;Kisspeptin/GnRH System