Human Reproduction, Journal Year: 2025, Volume and Issue: unknown
Published: May 13, 2025
Abstract STUDY QUESTION Which proteins are involved in the transition of human oocytes from germinal vesicle (GV) to metaphase I (MI) phase? SUMMARY ANSWER A total 2369 were identified, including 149 with significantly differential expression, 79 upregulated expression MI and 70 downregulated expression. WHAT IS KNOWN ALREADY During oocyte maturation, maternal RNA stored support early embryo development. However, GV matured vitro have a lower chance developing into blastocysts than oocytes. Therefore, identifying key differentially expressed between stages can provide better understanding development maturation mechanisms improve utilization DESIGN, SIZE, DURATION In total, 16 at collected female patients who underwent ovulation induction due male factor infertility requiring retrieval for ICSI. Differential identified using liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis, several was verified by immunofluorescence (IF). interference employed identify functions specific during maturation. PARTICIPANTS/MATERIALS, SETTING, METHODS immature discarded ICSI cycles (eight eight oocytes) 10 patients. Two cohorts zona pellucida removal, lysis, enzymatic digestion prior peptide detection LC-MS/MS methodology. Peptide outcomes subjected protein screening functional annotation employing distinct analytical algorithms datasets. To corroborate sequencing findings, exhibiting notable authenticated via IF. Concerning functionality, siRNA introduced phase, evaluated through observation polar body extrusion, alongside assessment efficacy IF analysis. MAIN RESULTS AND THE ROLE OF CHANCE Gene ontology analysis revealed that these mainly organic matter cell metabolism, biological regulation, primary nitrogen compound other processes. Kyoto Encyclopedia Genes Genomes genes following pathways: transport catabolism, signal transduction, folding, energy amino acid metabolism. The included actin-related 2 (ACTR2), NADH: Ubiquinone Oxidoreductase Core Subunit S1 (NDUFS1), Tubulin Gamma Complex Component 3 (TUBGCP3), Heat Shock Protein Family B (Small) Member 1 (HSPB1), Eukaryotic Translation Initiation Factor B, which mitochondrial function, division, transduction. ACTR2, HSPB1, NDUFS1, TUBGCP3 selected staining, difference fluorescence intensity consistent results. Three pairs primers designed each gene corresponding top (with siRNAs successfully seven proteins) study their results after interference. LARGE SCALE DATA See supplementary tables. LIMITATIONS, REASONS FOR CAUTION Although we some stage, crucial roles remain elusive. elucidate generated conditional knockout mice targeting proteins. WIDER IMPLICATIONS FINDINGS We conducted single-cell level stages. Our objective is ascertain potential supplementing culture medium augment both rates quality. FUNDING/COMPETING INTEREST(S) This research supported National Natural Science Foundation China (82171599 82471657, B.X., 82301871, L.L.); Postdoctoral (2024M763169, S.B.); Key Research Development Project (2029YFA0802600, B.X.). None authors has any conflict interest declare. TRIAL REGISTRATION NUMBER N/A.
Language: Английский