DNA甲基化参与山羊初情期启动的作用和机制

项目名称： DNA甲基化参与山羊初情期启动的作用和机制

项目编号： No.31472096

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 畜牧学与草地科学

项目作者： 方富贵

作者单位： 安徽农业大学

项目金额： 83万元

中文摘要： 初情期的早与迟直接影响动物繁殖性能和生产效益，目前其机制尚不完全清楚。近来研究报道DNA甲基化与初情期启动有关，抑制大鼠下丘脑DNA甲基化导致初情期失败，但作用机制仍不清楚。本项目以山羊为研究对象，拟利用多重免疫荧光标记等技术分析山羊在初情期前后下丘脑的总体DNA甲基化水平、DNA甲基转移酶及甲基化CpG结合蛋白的表达变化规律；采用亲和柱层析法富集高度甲基化组分并对其进行深度测序，绘制下丘脑全基因组DNA甲基化谱式，运用RNAseq技术绘制mRNA表达谱式，以期发现在初情期启动中起重要作用且其表达受到DNA甲基化状态调控的一批候选基因；通过亚硫酸氢盐测序等方法继而发现表达受到DNA甲基化调控的关键候选基因；从分子、细胞和动物水平阐明关键候选基因的功能和机制。综合上述结果，解析DNA甲基化参与山羊初情期启动的作用和机制，研究结果将有助于从新角度阐明动物初情期启动机制并为其调控提供新线索。

中文关键词： DNA甲基化；初情期；机制；山羊

英文摘要： The early and late onset of puberty is critical to reproductive performance and production payoff in animal. However, the mechanism underlying the onset of goat puberty is still poorly understood. Recently, DNA methylation was reported to be associated with the onset of puberty, which was failed in rats that were supressed in the hypothalamic DNA methylation. But no report on DNA methylation's role and its mechanism involved in the onset of puberty is given. In the present study, expression changes of global DNA methylation, DNA methyltransferases and methyl CpG binding proteins in hypothalamus of the prepubertal and pubertal goat will be analyzed by multiple immunofluorescence labeling. In order to find candidate genes being regulated by DNA methylation state and playing important role in the onset of puberty, a whole genome DNA methylation profile and the mRNA expression profile of the hypothalamus will be mapped by using affinity chromatography enrichment hypermethylation components and deep sequencing and RNAseq technology, respectively. Key candidate genes controlled by DNA methylation will be found by bisulfite sequencing PCR. The function and mechanism of key candidate genes will be clarified at the molecular, cellular and animal level. The role and mechanism of DNA methylation in the onset of puberty will be predicted according to the integrated above-mentioned research results. The results will be conducive to elucidate the mechanism of the onset of puberty in a novel perspective and provide new clues for the regulation of the onset of puberty.

英文关键词： DNA methylation;puberty;mechanism;goat