Discovery of Oogenesis Biomarkers from Mouse Oocytes Using a Single-Cell Proteomics Approach

Abstract

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.

On-capillary alkylation micro-reactor: a facile strategy for proteo-metabolome profiling in the same single cells

“…For example, Li et al for the first time identified 355 proteins from single mouse oocytes by employing the OAD chip. 17 Recently, an efficient and simplified single-cell proteomics (ES-SCP) workflow was developed to realize the identification of more than 1500 proteins from single oocytes, 29 which was a great advance in this field. More recently, Huang et al conducted multi-omics analysis of mouse oocytes, including single-cell proteomics, translatomics and transcriptomics, and detected 1200 proteins per single oocyte.…”

NUSAP1 regulates mouse oocyte meiotic maturation

“…Owing to recent advancements in mass spectrometry sensitivity, we conducted data mining and systematic comparisons of several known spindle assembly related genes 11,33–35 . At the mRNA expression level, consistent with these spindle‐related genes, NUSAP1 mRNA levels exhibited dominant expression in oocytes compared to granulosa cells (Figure S1).…”

Lectin-Based SP3 Technology Enables N-Glycoproteomic Analysis of Mouse Oocytes

“…Several N-linked glycoproteins in oocytes have been shown to be essential for sperm−egg interactions and fertility, including zona pellucida (ZP1−3), 2 Juno, 3 and Dpagt1. 4 However, the protein content in a single mouse oocyte is about 10−30 ng 5 and glycoproteomic analysis typically requires several hundred micrograms of protein for one experiment. Global mapping of the mouse oocyte N-glycoproteome has not been achieved and reported.…”