Proteomic and metabolomic insights into oxidative stress response activation in mouse embryos generated by

study questionHow different is the global proteomic and metabolic profile of mouse blastocysts generated by IVF, cultured in optimal (5% O SUMMARY ANSWER: We found that in IVF-generated embryos: (i) the proteome was more sensitive to high oxygen levels than the global metabolomic profile; (ii) enzymes involved in splicing and the spliceosome are altered; (iii) numerous metabolic pathways, particularly amino acids metabolism, are altered (iv) there is activation of the integrated stress response (ISR) and downregulation of mTOR pathways. WHAT IS KNOWN ALREADY: IVF culture conditions are known to affect the gene expression of embryos. However, comprehensive data on the global metabolic and proteomic changes that occur in IVF-generated embryos are unknown. STUDY DESIGN SIZE DURATION: Mouse embryos were generated by natural mating ( PARTICIPANTS/MATERIALS SETTING

methodsMouse blastocysts were collected from CD-1 and B6D2F1 strains as specified above. High-resolution liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used for proteomics, while high-performance liquid chromatography coupled with mass spectrometry (HILIC-MS) was used for metabolomics. In addition, Immunofluorescence was used to assess the activation of stress response pathways, including the ISR. MAIN RESULTS AND THE ROLE OF CHANCE: Proteomic analysis revealed significant changes in protein expression in embryos cultured under 20% O LARGE SCALE DATA: N/A. LIMITATIONS REASONS FOR CAUTION: Results in a murine model should be extrapolated with caution to human embryos. WIDER IMPLICATIONS OF THE

findingsThese findings offer valuable insights into how different IVF culture conditions, specifically oxygen levels, impact the global metabolic and proteomic profiles of embryos. These findings provide critical insights into the profound impact of IVF culture conditions, particularly oxygen levels, on the global metabolic and proteomic landscapes of embryos. By identifying key metabolic pathways disrupted by oxidative stress, we highlight the potential clinical importance of proteomic and metabolomic analyses in understanding embryo quality, improving ART, and ultimately enhancing pregnancy outcomes. The integration of metabolomic and proteomic data offers a comprehensive understanding of how oxidative stress influences cellular function. These insights have direct clinical relevance, providing a foundation for optimizing ART protocols to mitigate oxidative stress. STUDY FUNDING/COMPETING INTERESTS: This work was supported by grant R01 HD108166-01A1 from the National Institute of Child Health and Human Development (NICHD) to P.F.R. The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported.