四川大学学报（医学版）2025，Vol.56Issue(2)：419-424,6.DOI:10.12182/20250360603
小鼠次级卵母细胞和第一极体的高分辨率差异蛋白质组学分析
High-resolution Differential Proteomic Analysis of Mouse Secondary Oocytes and First Polar Bodies
摘要
Abstract
Objective To analyze the proteomic differences between mouse secondary oocytes,also known as metaphase Ⅱ oocytes(M Ⅱ),and the first polar bodies(PB1)using high-resolution single-cell proteomics,to identify key proteins regulating embryonic development,and to provide a molecular basis for optimizing in vitro oocyte maturation systems.Methods Paired samples of M Ⅱ(n=5)and PB1(n=5)were analyzed using high-resolution single-cell mass spectrometry(timsTOF HT).Quantitative proteomics and bioinformatics approaches were employed to conduct differential protein screening and functional enrichment.Results Using the timsTOF HT platform,we achieved the detection of over 3 000 proteins per single cell and identified 277 proteins specifically enriched in M Ⅱ.Gene Set Enrichment Analysis(GSEA)revealed that these M Ⅱ-specific proteins were significantly enriched in gene regulation and DNA damage repair pathways associated with mitochondrial energy metabolism.Cross-species GSEA comparison between human and mouse homologs demonstrated elevated expression of heat shock proteins,including Hsp90b1,Hspa5,etc.,in the mTORC1 pathway in M Ⅱ(P＜0.05).In addition,key factors regulating cumulus complex development,such as Calr,Aldoa,etc.,were significantly upregulated.Conclusion M Ⅱ strategically retains proteins essential for embryonic development through asymmetric division.The timsTOF HT platform demonstrated superior sensitivity in analyzing and identifying these proteins.According to the protein analysis results,the distribution of mTORC1 pathway proteins indicates that they play a key role in embryonic metabolism regulation.In particular,heat shock proteins facilitate protein folding and maintain endoplasmic reticulum homeostasis,thereby ensuring oocyte maturation and the embryonic developmental potential.关键词
单卵母细胞/蛋白质组学/极体/次级卵母细胞Key words
Single-cell oocyte/Proteomics/Polar body/Secondary oocyte引用本文复制引用
范晓丹,周黎明..小鼠次级卵母细胞和第一极体的高分辨率差异蛋白质组学分析[J].四川大学学报（医学版）,2025,56(2):419-424,6.基金项目
宁波市妇科疾病临床医学研究中心(No.2024L002)资助 （No.2024L002）
