H3K4me3调控RBP1的转录促进母猪初情启动的机制研究

母猪初情期是中等遗传力的重要经济性状，其遗传改良必能提高猪场的经济效益。项目申请人前期发现，H3K4me3可能调控RBP1启动子的染色质状态及其转录表达，影响卵巢颗粒细胞的增殖、视黄醇代谢、E2合成，进而促进母猪的卵泡成熟和初情启动。本项目拟在前期基础上，以初情正常和延缓猪的卵巢为实验材料，利用组织切片和高效液相色谱等方法，明确H3K4me3、RBP1基因、视黄醇代谢、E2合成、颗粒细胞增殖、卵泡成熟及初情启动之间的关系；然后，在细胞、组织和活体水平，采用CHART-PCR和CRISPR/Cas9等方法，研究H3K4me3调控RBP1启动子染色质状态及其转录表达，进而影响视黄醇代谢、E2合成、颗粒细胞功能、卵泡成熟和初情启动的作用机制。研究结果不仅能揭示H3K4me3调控RBP1的转录促进母猪初情启动的机制，还能为母猪初情期的遗传改良提供新的科学依据，在现代化的养猪生产中有极强的应用前景。

Puberty is one of the important economic traits with moderate heritability in gilts, and its genetic improvement will make for a favorable economic effect on the modern commercial pig farms. Our previous results revealed that H3K4me3 might regulate the chromatin remodeling of Retinol-binding protein 1 (RBP1) promoter to control the transcription of RBP1, to promote the proliferation of ovarian granulosa cells, retinol metabolism, and estrogen synthesis, and to advance the follicular maturation and the initiation of puberty in gilts. Therefore, in this project, using histological sections, high performance liquid chromatography, chromatin assay by RT-PCR with DNase I and CRISPR/Cas9, we first try to identify the relationship among H3K4me3, RBP1 gene, retinol metabolism estrogen synthesis, cell proliferation, follicular maturation, and the initiation of puberty by utilizing the ovaries from the normal and delayed gilts. Then we explore and determine the mechanism that H3K4me3 regulates the chromatin remodeling of RBP1 promoter and the transcription of RBP1 to promote the proliferation of ovarian granulosa cells, retinol metabolism, and estrogen synthesis to advance the follicular maturation and the initiation of puberty in the porcine ovarian granulosa cell, cultured follicles and animal model. These works will not only figure out the mechanism of H3K4me3 regulating the transcription of RBP1 to advance the initiation of puberty in gilts, but also improve the theoretical resource available for genetic improvement of puberty in gilts, and these works will harbor a highly applicable prospects in the modern commercial pig farms.