Guo Yueshuai, Gao Mengmeng, Liu Xiaofei, Zhang Haotian, Wang Yue, Yan Tong, Wang Bing, Han Xudong, Qi Yaling, Zhu Hui, Situ Chenghao, Li Yan, Guo Xuejiang
State Key Laboratory of Reproductive Medicine and Offspring Health, Department of Histology and Embryology, Nanjing Medical University, Nanjing, China.
State Key Laboratory of Reproductive Medicine and Offspring Health, Department of Histology and Embryology, Nanjing Medical University, Nanjing, China; School of Medicine, Southeast University, Nanjing, China.
Mol Cell Proteomics. 2025 Jan;24(1):100882. doi: 10.1016/j.mcpro.2024.100882. Epub 2024 Nov 20.
Once ovulated, the oocyte has to be fertilized in a short time window or it will undergo post-ovulation aging (POA), whose underlying mechanisms are still not elucidated. Here, we optimized single-cell proteomics methods and performed single-cell transcriptomic, proteomic, and phosphoproteomic analysis of fresh, POA, and melatonin-treated POA oocytes. POA oocytes showed downregulation of most differentially expressed proteins, with little correlation with mRNA expression, and the protein changes can be rescued by melatonin treatment. MG132 treatment rescued the decreased fertilization and polyspermy rates and upregulated fragmentation and parthenogenesis rates of POA oocytes. MG132-treated oocytes displayed health status at proteome, phosphoproteome, and fertilization ability similar to fresh oocytes, suggesting that protein stabilization might be the underlying mechanism for melatonin to rescue POA. The important roles of proteasome-mediated protein degradation during oocyte POA revealed by single-cell multi-omics analyses offer new perspectives for increasing oocyte quality during POA and improving assisted reproduction technologies.
一旦排卵，卵母细胞必须在短时间内受精，否则它将经历排卵后老化（POA），其潜在机制仍未阐明。在这里，我们优化了单细胞蛋白质组学方法，并对新鲜、POA和褪黑素处理的POA卵母细胞进行了单细胞转录组学、蛋白质组学和磷酸蛋白质组学分析。POA卵母细胞中大多数差异表达蛋白下调，与mRNA表达相关性很小，而褪黑素处理可以挽救蛋白质变化。MG132处理挽救了POA卵母细胞受精率和多精入卵率的下降，并上调了碎片化和孤雌生殖率。MG132处理的卵母细胞在蛋白质组、磷酸蛋白质组和受精能力方面显示出与新鲜卵母细胞相似的健康状态，这表明蛋白质稳定可能是褪黑素挽救POA的潜在机制。单细胞多组学分析揭示的蛋白酶体介导的蛋白质降解在卵母细胞POA过程中的重要作用，为提高POA期间的卵母细胞质量和改进辅助生殖技术提供了新细胞多组学分析揭示的蛋白酶体介导的蛋白质降解在卵母细胞POA过程中的重要作用，为提高POA期间的卵母细胞质量和改进辅助生殖技术提供了新的视角。
