State Key Laboratory of Plant Environmental Resilience, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
State Key Laboratory of Farm Animal Biotech Breeding, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
J Proteome Res. 2023 Jun 2;22(6):2067-2078. doi: 10.1021/acs.jproteome.3c00157. Epub 2023 May 8.
We established an efficient and simplified single-cell proteomics (ES-SCP) workflow to realize proteomics profiling at the single-oocyte level. With the ES-SCP workflow, we constructed a deep coverage proteome library during oocyte maturation, which contained more than 6000 protein groups, and identified and quantified more than 4000 protein groups from a pool of only 15 oocytes at germinal vesicle (GV), GV breakdown (GVBD), and metaphase II (MII) stages. More than 1500 protein groups can be identified from single oocytes. We found that marker proteins including maternal factors and mRNA regulators, such as ZAR1, TLE6, and BTG4, showed significant variations in abundance during oocyte maturation, and it was discovered that maternal mRNA degradation was indispensable during oocyte maturation. Proteomics analysis from single oocytes revealed that changes in antioxidant factors, maternal factors, mRNA stabilization, and energy metabolism were the factors that affect the oocyte quality during ovary aging. Our data laid the foundation for future innovations in assisted reproduction.
我们建立了一种高效简化的单细胞蛋白质组学（ES-SCP）工作流程，以实现单细胞水平的蛋白质组学分析。通过 ES-SCP 工作流程，我们在卵母细胞成熟过程中构建了一个深度覆盖的蛋白质组文库，其中包含超过 6000 个蛋白质组，并且能够从仅 15 个处于生发泡（GV）、GV 破裂（GVBD）和中期 II（MII）阶段的卵母细胞中鉴定和定量超过 4000 个蛋白质组。从单个卵母细胞中可以鉴定出超过 1500 个蛋白质组。我们发现，包括母体因子和 mRNA 调节剂在内的标记蛋白，如 ZAR1、TLE6 和 BTG4，在卵母细胞成熟过程中丰度有明显变化，并且发现母源 mRNA 降解在卵母细胞成熟过程中是必不可少的。来自单个卵母细胞的蛋白质组学分析表明，抗氧化因子、母体因子、mRNA 稳定和能量代谢的变化是影响卵巢衰老过程中卵母细胞质量的因素。我们的数据为辅助生殖领域的未来创新奠定了基础。
