TNF-α对同期发情-排卵山羊卵母细胞老化的影响及机制

以同期排卵和定时输精技术为核心的批次化生产，能够大幅度提高母羊的繁殖力。然而，该技术面临母羊经外源激素处理后卵母细胞发育和受精异常导致受胎率下降的问题，阻碍了该技术的推广应用。申请人前期研究发现在不同季节及应用不同方法处理，山羊卵母细胞的发育率和受精率有差异。利用小鼠研究发现，肿瘤坏死因子（TNF-α）促进了卵母细胞的老化，并影响了其发育和受精。TNF-α对山羊卵母细胞的影响及作用机制未见报道。本项目以山羊为研究对象，应用B超监测、免疫荧光和蛋白印迹及细胞水平敲低等技术，探索外源激素处理过程中TNF-α对同期发情-排卵山羊卵母细胞的细胞周期、老化、发育和受精的影响以及卵母细胞中TNF-R1的表达、微管变化和早期胚胎发育情况。项目完成后，将明晰TNF-α对同期发情-排卵山羊卵母细胞老化的调控作用机制，为相关繁殖调控技术研发和提高卵母细胞发育能力提供理论参考。

The batch production based on the technology of ovulation synchronization-timed artificial insemination（OS-TAI）can greatly improve the fecundity of goat. However, this technique is faced with the problem of the decrease of goat pregnancy rate via abnormal oocyte development and fertilization by using exogenous hormone, which hinders the large scale practical application of OS-TAI. In our previous studies, we found that the development and fertilization rates of goat oocytes were different in different seasons and with different treatments. Using mouse models, the applicant found that tumor necrosis factor（TNF-α）promoted aging of mouse oocytes, and affected their development and fertilization. The effect and mechanism of TNF-α on goat oocytes aging have not well understood. Based on these evidences, the subject will be goat as the research object, using B-mode ultrasound monitoring, immunofluorescence staining, western blotting and knockdown in cell level, to explore the influence of TNF-α for the cell cycle, aging, development, fertilization, expression of TNF-R1, variation of microtubules in oocyte, and early embryonic development during the goat synchronization of estrus and ovulation with exogenous hormones. After the completion of the project, the effect and mechanism of TNF-α regulating the aging of oocytes will be clarified during the synchronization of estrus and ovulation in goats, and these can also provide theoretical reference for improving the research and development of related reproductive regulation technologies and the development ability of oocytes in practice.