Li Rongqing, Zhang Wenjia, Gao Yuanshuai, Xie Zhiqiang, He Jiangfeng, Fang Qinyuan, El-Sherbiny Mohamed, Gao Min, Wang Zheng, Zhang Teng, Liu Fang, Wang Biao, Liu Yongbin
Key Laboratory of Herbivorous Livestock Reproductive Regulation, National Sheep Genetic Evaluation Center, Inner Mongolia University, Hohhot 010030, China.
Inner Mongolia Academy of Agricultural & Animal Husbandry Sciences, Hohhot 010031, China.
Animals (Basel). 2025 Aug 11;15(16):2354. doi: 10.3390/ani15162354.
Normal proliferation of ovarian granulosa cells is essential for follicular development. The results of this study showed that ADAMTS1 was primarily localized in the cytoplasm of granulosa cells in sheep ovarian follicles, as revealed by immunohistochemistry and immunofluorescence staining. Knockdown and overexpression experiments of in granulosa cells demonstrated that the number of EdU-positive cells significantly decreased in the knockdown group ( < 0.05), while the expression levels of ( < 0.05), / ( < 0.01), and ( < 0.05) were significantly upregulated, indicating that knockdown of markedly inhibited granulosa cell proliferation. In contrast, overexpression of significantly promoted cell proliferation. Transcriptome sequencing revealed that and were significantly downregulated in the knockdown group and significantly upregulated in the overexpression group, which was confirmed by Quantitative Polymerase Chain Reaction (Q-PCR) ( < 0.05). KEGG enrichment analysis showed that was significantly enriched in the glycine, serine and threonine metabolism and vitamin B6 metabolism pathways. Molecular docking analysis indicated a stable binding interface between ADAMTS1 and PSAT1. Based on these findings, we speculate that may regulate amino acid metabolism in ovarian granulosa cells by modulating the expression of SLC6A9, which in turn affects in the glycine, serine, and threonine metabolism and vitamin B6 metabolism pathways, thereby influencing granulosa cell proliferation.
卵巢颗粒细胞的正常增殖对卵泡发育至关重要。本研究结果表明，通过免疫组织化学和免疫荧光染色显示，ADAMTS1主要定位于绵羊卵巢卵泡颗粒细胞的细胞质中。颗粒细胞的敲低和过表达实验表明，敲低组中EdU阳性细胞数量显著减少（<0.05），而（<0.05）、/（<0.01）和（<0.05）的表达水平显著上调，表明敲低显著抑制颗粒细胞增殖。相反，过表达显著促进细胞增殖。转录组测序显示，敲低组中显著下调，过表达组中显著上调，这通过定量聚合酶链反应（Q-PCR）得到证实（<0.05）。KEGG富集分析表明，在甘氨酸、丝氨酸和苏氨酸代谢以及维生素B6代谢途径中显著富集。分子对接分析表明ADAMTS1与PSAT1之间存在稳定的结合界面。基于这些发现，我们推测可能通过调节SLC6A9的表达来调节卵巢颗粒细胞中的氨基酸代谢，进而影响甘氨酸、丝氨酸和苏氨酸代谢以及维生素B6代谢途径中的，从而影响颗粒细胞增殖。
