双峰驼精清诱导排卵的分子调控机理

The population of bactrian camel has a dramatic decline in recent years, the main reasons caused this situation is the lower reproductive performance, longer growth period and a slower growth rate, especially the completely unclear molecular regulation mechanism of bactrian camel seminal plasma induced ovulation. Early studies of our tern find and reported that the bactrian camel is a kind of semen induced ovulation animal. The female bactrian camel will begin to ovulation after intramuscular injecting seminal plasma for 24 hours, complete ovulation for 36-48 hours. Whole genome sequencing and proteomics approach were implemented in this project for screening key target genes and proteins in induced ovulation, verified the expression pattern and constructed the network of candidate genes and proteins based on our previous study. And further clarified the molecular regulation mechanism of bactrian camel seminal plasma induced ovulation. Which will provid the theoretical basis for bactrian camel and other induced ovulation animals in breeding and reproduction related diseases.

近年来双峰驼数量急剧下降，其主要原因是双峰驼繁殖性能低、周期长和速度慢，特别是双峰驼排卵的分子机理完全不清楚。课题组早期发现并报道双峰驼为精液诱导排卵动物，母驼肌肉注射精清24h后即开始排卵，36-48h完成排卵。在此基础上，本项目通过全基因组重测序和蛋白质组学的方法筛选诱导排卵过程中关键的靶基因和蛋白，并验证其表达规律，构建其基因-蛋白互调控作网络，进一步解析双峰驼精清诱导排卵的分子机理。为今后双峰驼和诱导性排卵动物繁殖及生殖相关疾病的研究提供依据。

双峰驼(Camelus Bactrianus)作为西北地区特有的濒危物种，也是荒漠和半荒漠草原特有的优势畜种资源，具有重要的经济和科研价值。由于双峰驼是严格的季节性发情及诱导排卵动物，其生殖周期长、繁殖性能低、生长速度慢，迄今未见双峰驼生殖相关候选基因和功能蛋白等相关报道，并且双峰驼诱导排卵的分子机制仍不清楚，分子调控机理完全空白。故而，本研究首先从繁殖期和非繁殖期公驼入手，通过蛋白质组学筛选与生殖发育相关的血清差异蛋白，成功筛选32个差异蛋白，其中5个差异蛋白与季节性发情及生殖激素相关（PCGF5、H1.2、ANTXR2、FOLR1和RBP4），并通过体外细胞模型验证差异蛋白的功能。其次，利用肌肉注射精清诱导排卵，利用蛋白质组学成功筛选出404个卵巢差异蛋白，其中NR5A1,FST，PAK1，PRL及CAMK2G为关键功能蛋白，并对其进行性腺轴表达的功能验证，同时对CAMK2G的功能进行了体外细胞模型的验证。通过以上内容的研究，从蛋白质组学筛选双峰驼季节性发情及精清诱导排卵的关键蛋白；深入揭示双峰驼季节发情及精清诱导排卵的分子调控。为双峰驼的繁殖和育种提供参考依据，为诱导性排卵动物繁殖及其生殖相关疾病的研究提供理论基础。