绵羊内源性逆转录病毒调节胚胎滋养层细胞融合发育的机制

前期的研究证实绵羊内源性逆转录病毒（enJSRV）及其受体HYAL2在不同妊娠期的胎盘和孕体中的表达变化规律均与绵羊的妊娠过程和孕体的发育密切相关。胎盘的发生是一个受到高度调控的动态生理过程，关键环节是滋养层细胞的分化。本项目的提出是在原有研究的基础上通过构建稳定表达enJSRV的Env蛋白及其受体HYAL2的293T细胞系，然后进行共培养观察细胞融合机制并通过RNA干扰诱导enJSRV-env基因沉默，构建并筛选最佳干扰质粒转染原代培养的滋养层细胞，检测细胞生长发育情况，采用免疫沉淀法检测滋养层细胞分化发育中MAPK信号传导通路的调节。同时将构建的超表达质粒和干扰质粒包装慢病毒感染妊娠小鼠，观察小鼠妊娠过程和胚胎发育情况，进一步验证体外细胞中的实验结果，从而揭示enJSRV在胚胎发育特别是滋养层细胞融合分化过程的调控机制。研究成果将对动物的生存、发育、繁殖等具有重要的应用价值和理论意义。

Previous research confirmed that the expression of endogenous Jaagsiekte retrovirus (enJSRV) and its receptor Hyaluronidase-2 (Hyal-2) in placenta and conceptus is closely related to the pregnancy and development process. the formation of the placenta is a dynamic physiological process highly regulated by,and trophoblast cells differentiation is the key to placentogenesis. On the basis of these earlier studies, the project proposed is designed to establish 293T cell line stably expressing the enJSRV envelope protein (Env) and Hyal-2 .Then co-culture them and observe the fusion of subsequent cells. Also we explore RNA interference aiming to induce enJSRV-env gene silencing, observe the growth of the primarily cultured trophoblast cells by transfected with the best RNAi plasmid and evaluate the regulatory role of MAPK signal transduction pathway in trophoblast cells differentiation by the technique of immune-precipitation. Package eukaryotic expression vector and RNAi plasmid into the recombinant lentivirus to infect the pregnancy BALB/c mice and observe the pregnancy process and embryo development so as to confirm the in vitro experimental results further which investigate the regulation role of enJSRV in embryo development , especially in trophoblast cells fusion and differentiation process . The research results will provide deep insight into the research of animal survival, development, reproduction and other aspects.