IFN-τ调控奶牛子宫内膜上皮细胞差异表达BoLA-I的分子机制

项目名称： IFN-τ调控奶牛子宫内膜上皮细胞差异表达BoLA-I的分子机制

项目编号： No.31472254

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 邓干臻

作者单位： 华中农业大学

项目金额： 88万元

中文摘要： BoLA-I在牛妊娠免疫过程中起决定作用。IFN-τ在围着床期由胎盘滋养层细胞分泌，调节EEC差异表达BoLA-I，继而调节母-胎界面免疫。本项目构建IFN-τ调节奶牛子宫内膜上皮细胞差异表达BoLA-I的体外细胞模型，采用Illumina深度测序技术分析EEC中差异表达的miRNA并构建miRNA表达谱及其靶向关系，利用qRT-PCR、Northern blot、原位杂交、生物信息学、miRNA-mRNA复合物、反义寡核苷酸、超表达技术、报告基因等技术鉴定和验证靶向关系及其功能；利用超表达和RNAi技术研究上游miRNA/mRNA功能，综合判断miRNA及上游miRNA/mRNA在EEC差异表达BoLA-I中的作用。项目将发现并验证相关miRNA表达谱和BoLA-I上游miRNAs/mRANs及其功能，阐明IFN-τ调节EEC差异表达BoLA-I的分子机理，为提升奶牛繁殖力提供科学依据。

中文关键词： 奶牛；IFN-τ；BoLA-I；子宫内膜上皮细胞；分子机制

英文摘要： BoLA-I take major contribution to maternal immune response to the semiallogenic fetus.Embryo trophoblastic cells only express IFN-τ at the period of preimplantation, and its receptor is located on endometrial epitherial cells(EEC). IFN-τ differentially mediates the expression of BoLA-I on EEC, and then regulate the immune response on the interface of maternal-fetus,and therefore make the formation of endometrial condition for implantation. To test the effect of miRNA and the upstream miRNAs/mRNAs on the differential expression of BoLA-I on EEC mediated with IFN-τ, we construct the in vitro model of IFN-τ regulating the differential expression of BoLA-I on EEC. Based on the theory of miRNA/mRNA,Illumina deep sequensing will be adapted to analyze the miRNAs spectrum on the above cell model. qRT-PCR, Northern blot,in situ hydridization will be used for the confirmation of miRNAs. Bioimfornatics will applied to predict the miRNA/mRNA what is identified with miRNA/mRNA compound analytic precess and pGL3 Luciferase report genetic carrier. Antisense oligonucleotid and miRNA overexpression methods will be adapted to test the function of special miRNA.miRNA interference and overexpression induced with retrovirus will be used to analyze the regulating effect of upstream genes ahead BoLA-I. This project will discove and identify the miRNA spectrum and the upstream miRNAs/mRNAS between IFN-τ with BoLA-I in EEC of dairy cows, and then illuminate the molecular mechanism of IFN-τ differentially regulating BoLA-I expression, which should be essential to promote the productivity of dairy cows.

英文关键词： Dairy Cow;IFN-τ;BoLA-I;Endometrial epithelium cells;Molecular mechanism