Result-X101SC25013530_Z02_J001_B1_43
Description
This dataset contains quantitative proteomic profiles of bovine cumulus cells collected at key stages of in vitro oocyte maturation. The purpose of this study was to investigate the dynamic protein expression changes that occur in cumulus cells during the transition from the germinal vesicle (GV) stage to the metaphase II (MII) stage, providing insight into the molecular mechanisms that support oocyte maturation. Cumulus–oocyte complexes (COCs) were aspirated from bovine ovaries and cultured under standard in vitro maturation (IVM) conditions. Cumulus cells were isolated at GV (0 h) and MII (24 h) time points. Proteomic analysis was performed using high-resolution liquid chromatography–tandem mass spectrometry (LC–MS/MS). Protein identification and quantification were achieved through database-dependent searches, and label-free quantification was applied to determine relative protein abundance across biological replicates. The dataset includes raw mass spectrometry files, processed protein identification and quantification tables, and experimental metadata. These data provide a resource for understanding the proteomic remodeling of cumulus cells during oocyte maturation and may serve as a reference for studies on mammalian follicular development, oocyte competence, and assisted reproductive technologies.
Files
Steps to reproduce
Cumulus–oocyte complexes (COCs) were collected from bovine ovaries, and cumulus cells were isolated at germinal vesicle (GV, 0 h) and metaphase II (MII, 24 h) stages (3 biological replicates per stage, ~150 oocytes per replicate). Total proteins were extracted using DB lysis buffer (6 M urea, 100 mM TEAB, pH 8.5) followed by ultrasonication and centrifugation. Supernatants were reduced with DTT, alkylated with iodoacetamide, and quantified using the Bradford assay. Proteins were digested with trypsin, and peptides were labeled with tandem mass tag (TMT) reagents, pooled, desalted, and fractionated by high-pH reverse-phase HPLC. Fractions were dried, reconstituted in formic acid, and analyzed by nano-UHPLC coupled to a Q Exactive HF-X Orbitrap mass spectrometer (Thermo Fisher Scientific). Peptides were separated on in-house packed C18 columns, and MS data were acquired in data-dependent acquisition mode. Raw spectra were processed using Proteome Discoverer (Thermo Scientific) against the Bos taurus UniProt database. Search parameters included a precursor mass tolerance of 10 ppm and fragment mass tolerance of 0.02 Da. Carbamidomethylation of cysteine was set as a fixed modification, while oxidation (M) and TMT plex were variable modifications. Up to two missed cleavages were allowed. Protein and peptide identifications were filtered at 1% FDR. Quantitative comparisons between GV and MII cumulus cells were performed using t-tests, with differentially expressed proteins (DEPs) defined as p < 0.05 and |log2FC| > 1. Functional annotation of proteins and DEPs was conducted using GO, InterPro, COG, KEGG pathway analysis, and STRING-db for protein–protein interaction predictions.
Institutions
- Zhejiang University