雌激素与小鼠植入前胚胎发育过程中表观遗传模式正确建立的相关性研究

项目名称： 雌激素与小鼠植入前胚胎发育过程中表观遗传模式正确建立的相关性研究

项目编号： No.31501906

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 动物学

项目作者： 吴风瑞

作者单位： 阜阳师范学院

项目金额： 19万元

中文摘要： 雌激素和表观遗传修饰对小鼠植入前胚胎发育有重要影响，但鲜有揭示二者在胚胎发育过程中的相关性特别是雌激素功能的报道。结合前期研究，我们提出雌激素可能通过影响植入前胚胎表观遗传模式的建立和发育相关因子的表达，参与胚胎发育的观点，并拟以小鼠体外胚和体细胞克隆胚等植入前胚胎为材料，从以下3方面论证我们的设想：1）在培养基中添加雌激素，利用qPCR、免疫荧光、Western杂交、酶活力分析等方法，研究雌激素对表观遗传修饰酶、重要发育基因表达和酶活力的影响；2）通过启动子分析、ChIP和EMSA等方法揭示雌激素对表观遗传修饰相关酶基因转录调控的分子机制；3）通过RNA干扰抑制ER基因表达和以ER基因敲除的稳转系细胞为供体构建克隆胚，在体验证雌激素对表观遗传模式正确建立的影响和相关性。本研究将阐明雌激素在小鼠植入前胚胎发育过程中功能和作用机制，为提高克隆胚发育率和实验动物进一步模式化奠定基础。

中文关键词： 雌激素；表观遗传修饰；转录调控；分子机制；小鼠

英文摘要： Previous studies suggested that estrogen and epigenetic modification play important roles in mouse preimplantation embryo development and implantation, but the correlation between estrogen and the establishment of epigenetic modification pattern in this process remains unclear, especially the function of estrogen. Based on our previous studies, we propose that estrogen control the epigenetic modification, such as DNA methylation and histone modification, through modulating the transcription regulation of epigenetic modifying enzyme genes to participate in mouse preimplantation embryo development. Using culture system with estrogen, promoter assay system, real-time PCR, immunofluorescence microscopy, Western blot and enzyme activity analysis, we will examine the effect of estrogen on the mRNA expression, protein, and promoter activities of epigenetic modifying enzyme genes and enzyme activity in mouse IVF and cloned embryos. Electrophoretic Mobility Shift Assay (EMSA), Chromatin Immunoprecipitation(ChIP) and RNAi technology will be further performed to illustrate the molecular mechanisms of transcription regulation of epigenetic modifying enzymes regulation by estrogen and its receptors. Furthermore, ER gene knockout cells-cloned embryo was constructed to further in vivo validate the effect of estrogen on the cloned embryo development and epigenetic modification. The study aims to clarify the correlation between estrogen and the establishment of flawless epigenetic pattern during mouse preimplantation embryo development, and the results will provide new insight into the function of estrogen and epigenetic modification on the embryo development, and basis for promoting the rate of cloned embryo development and assisted reproduction.

英文关键词： Estrogen;Epigenetic modification;Transcription regulation;Molecular mechanism;Mouse