山羊卵泡选择中miRNAs的参与及其调控机制研究

项目名称： 山羊卵泡选择中miRNAs的参与及其调控机制研究

项目编号： No.31301944

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 农业科学

项目作者： 娜日苏

作者单位： 西南大学

项目金额： 24万元

中文摘要： 卵泡选择（follicle selection）是决定排卵数量的重要步骤，属于多因素参与调节的复杂生命过程，其机制尚未明确。探索miRNAs参与卵泡选择调控的分子机制是对卵泡选择调控机理的有益补充和完善，也对山羊排卵机理研究及卵泡储备资源开发具有重要的意义。本项目以山羊卵泡为研究材料，利用山羊卵巢表达的miRNAs（前期基础）制定miRNA芯片寻找募集卵泡、优势卵泡及从属卵泡中差异表达的miRNAs，通过生物信息学手段预测miRNA的靶基因，进一步利用荧光素酶报告系统明确miRNA与靶基因的调控关系；在此基础上，利用FSH诱导颗粒细胞增殖研究FSH与miRNAs的调控关系,利用miRNA抑制剂（和类似物）体内外干扰途径研究miRNA及其靶基因调控颗粒细胞增殖、激素合成及对卵泡发育波动力学的影响，阐明miRNAs参与卵泡选择的调控机制。

中文关键词： 卵泡；山羊；微小RNA；原始卵泡；初级卵泡

英文摘要： The number of ovulations at each ovarian cycle is determined by a tightly controlled follicle selection process. The mechanism underlying follicle selection take place on different scales and different levels, how precisely the follicular selection is controlled remains a critical and important question in reproductive biology. It is a supplement and improvement of the follicle selection mechanism by studying the involvement and regulation of miRNAs in goat follicular selection , it also provides important values to goat ovulation research and follicular resources development. In this study, we employ miRNA microarray using miRNAs expressed in goat ovaries to evaluate the differently expressed miRNAs between pre-selected growing antral follicle, dominant follicle and subordinate one. Some of these differentially expressed miRNAs are validated using quantitative PCR assay. Top significant GO categories, biological functions and different canonical pathways are analyzed for the specific targets of differentially expressed miRNAs. Using luciferase reporter system to validate target genes. Inducing granulosa cell proliferation by FSH to identify the regulation between FSH and miRNAs. Using miRNA inhibitors or locked nucleic acid oligonucleotides complementary to specific miRNAs to identify the function of miRNAs on

英文关键词： follicle；goat；microRNA；primordial follicle；primary follicle