Transcriptome Analysis Reveals the Molecular Mechanisms by Which ADAMTS1 Influences the Proliferation of Ovarian Granulosa Cells in Sheep

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 <i>ADAMTS1</i> in granulosa cells demonstrated that the number of EdU-positive cells significantly decreased in the knockdown group (<i>p</i> < 0.05), while the expression levels of <i>Bax</i> (<i>p</i> < 0.05), <i>Bax</i>/<i>Bcl2</i> (<i>p</i> < 0.01), and <i>caspase3</i> (<i>p</i> < 0.05) were significantly upregulated, indicating that knockdown of <i>ADAMTS1</i> markedly inhibited granulosa cell proliferation. In contrast, overexpression of <i>ADAMTS1</i> significantly promoted cell proliferation. Transcriptome sequencing revealed that <i>PSAT1</i> and <i>SLC6A9</i> were significantly downregulated in the knockdown group and significantly upregulated in the overexpression group, which was confirmed by Quantitative Polymerase Chain Reaction (Q-PCR) (<i>p</i> < 0.05). KEGG enrichment analysis showed that <i>PSAT1</i> 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 <i>ADAMTS1</i> may regulate amino acid metabolism in ovarian granulosa cells by modulating the expression of SLC6A9, which in turn affects <i>PSAT1</i> in the glycine, serine, and threonine metabolism and vitamin B6 metabolism pathways, thereby influencing granulosa cell proliferation.