LNC_005531介导chi-miR-2290靶向WNT4基因调控山羊排卵的机理

排卵是山羊产羔的重要环节，然而调控山羊排卵的分子机制尚未明晰。申请人前期发现WNT4与山羊多排卵数显著相关，且初步证实chi-miR-2290、LNC_005531是WNT4的调控元件。因此，提出“LNC_005531介导chi-miR-2290靶向WNT4基因调控山羊排卵的机理”这一科学假设。本研究首先在细胞水平过表达和干扰试验精确探究LNC_005531介导chi-miR-2290靶向调控WNT4基因的转录表达模式；其次，分离培养不同卵泡发育时期的颗粒细胞，测定其增殖、分化指标及激素分泌，解析LNC_005531-chi-miR-2290-WNT4通路的不同调控模式；最后，分析山羊排卵标志因子PR、COX转录表达及激素分泌变化，阐明LNC_005531介导chi-miR-2290靶向WNT4基因调控山羊排卵的作用机制。本项目将为筛选山羊排卵性状主效基因和分子设计育种提供新靶点。

Ovulation is an important part of kidding in goats. However, the molecular mechanisms regulating ovulation in goats are not yet understood. In our previous study, we found that WNT4 was significantly associated with the number of ovulation in goats, and we preliminarily confirmed that chi-miR-2290 and LNC_005531 are the regulatory elements of WNT4. Therefore, we propose the scientific hypothesis that "LNC_005531 mediates the mechanism of chi-miR-2290 targeting WNT4 gene to regulate ovulation in goats". In this study, we firstly investigated the regulation mode of LNC_005531 mediated chi-miR-2290 targeted WNT4 gene expression through cell overexpression and interference assays. Secondly, we analyzed the effect of LNC_005531-chi-miR-2290-WNT4 pathway through cell proliferation, differentiation marker genes and hormone assay in granulosa cells cultured in vitro at different growth stages. Finally, the mechanism of LNC_005531 mediated chi-miR-2290 targeting WNT4 gene to regulate the ovulation process in goats was elucidated by transcriptional expression of ovulation markers PR and COX and hormone assays. This project will provide new targets for screening the major genes of ovulation in goats and molecular design breeding.