Integrated ultrasensitive metabolomics and single-cell transcriptomics identify crucial regulators of sheep oocyte maturation and early embryo development in vitro
Introduction: Developmental competence of oocytes matured in vitro is limited due to a lack of complete understanding of metabolism and metabolic gene expression during oocyte maturation and embryo development. Conventional metabolic analysis requires a large number of samples and is not efficiently...
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Elsevier
2025-07-01
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| Series: | Journal of Advanced Research |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2090123224003813 |
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| author | Bo Pan JianPeng Qin KunLin Du LuYao Zhang GongXue Jia JiangFeng Ye QiuXia Liang QiEn Yang GuangBin Zhou |
| author_facet | Bo Pan JianPeng Qin KunLin Du LuYao Zhang GongXue Jia JiangFeng Ye QiuXia Liang QiEn Yang GuangBin Zhou |
| author_sort | Bo Pan |
| collection | DOAJ |
| description | Introduction: Developmental competence of oocytes matured in vitro is limited due to a lack of complete understanding of metabolism and metabolic gene expression during oocyte maturation and embryo development. Conventional metabolic analysis requires a large number of samples and is not efficiently applicable in oocytes and early embryos, thereby posing challenges in identifying key metabolites and regulating their in vitro culture system. Objectives: To enhance the developmental competence of sheep oocytes, this study aimed to identify and supplement essential metabolites that were deficient in the culture systems. Methods: The metabolic characteristics of oocytes and embryos were determined using ultrasensitive metabolomics analysis on trace samples and single-cell RNA-seq. By conducting integrated analyses of metabolites in cells (oocytes and embryos) and their developmental microenvironment (follicular fluid, oviductal fluid, and in vitro culture systems), we identified key missing metabolites in the in vitro culture systems. In order to assess the impact of these key missing metabolites on oocyte development competence, we performed in vitro culture experiments. Furthermore, omics analyses were employed to elucidate the underlying mechanisms. Results: Our findings demonstrated that betaine, carnitine and creatine were the key missing metabolites in vitro culture systems and supplementation of betaine and L-carnitine significantly improved the blastocyst formation rate (67.48% and 48.61%). Through in vitro culture experiments and omics analyses, we have discovered that L-carnitine had the potential to promote fatty acid oxidation, reduce lipid content and lipid peroxidation level, and regulate spindle morphological grade through fatty acid degradation pathway. Additionally, betaine may participate in methylation modification and osmotic pressure regulation, thereby potentially improving oocyte maturation and early embryo development in sheep. Conclusion: Together, these analyses identified key metabolites that promote ovine oocyte maturation and early embryo development, while also providing a new viewpoint to improve clinical applications such as oocyte maturation or embryo culture. |
| format | Article |
| id | doaj-art-0bd610e30d3b407d85833b864faad8df |
| institution | OA Journals |
| issn | 2090-1232 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Advanced Research |
| spelling | doaj-art-0bd610e30d3b407d85833b864faad8df2025-08-20T02:36:46ZengElsevierJournal of Advanced Research2090-12322025-07-017314716010.1016/j.jare.2024.08.040Integrated ultrasensitive metabolomics and single-cell transcriptomics identify crucial regulators of sheep oocyte maturation and early embryo development in vitroBo Pan0JianPeng Qin1KunLin Du2LuYao Zhang3GongXue Jia4JiangFeng Ye5QiuXia Liang6QiEn Yang7GuangBin Zhou8State Key Laboratory of Swine and Poultry Breeding Industry, Key Laboratory of Livestock and Poultry Multiomics, Ministry of Agriculture and Rural Affairs, Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, College of Animal Science and Technology, Sichuan Agricultural University, Sichuan, Chengdu 611130, PR ChinaState Key Laboratory of Swine and Poultry Breeding Industry, Key Laboratory of Livestock and Poultry Multiomics, Ministry of Agriculture and Rural Affairs, Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, College of Animal Science and Technology, Sichuan Agricultural University, Sichuan, Chengdu 611130, PR ChinaState Key Laboratory of Swine and Poultry Breeding Industry, Key Laboratory of Livestock and Poultry Multiomics, Ministry of Agriculture and Rural Affairs, Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, College of Animal Science and Technology, Sichuan Agricultural University, Sichuan, Chengdu 611130, PR ChinaKey Laboratory of Adaptation and Evolution of Plateau Biota, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Qinghai, Xining 810001, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China; Qinghai Key Laboratory of Animal Ecological Genomics, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Qinghai, Xining 810001, PR ChinaKey Laboratory of Adaptation and Evolution of Plateau Biota, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Qinghai, Xining 810001, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China; Qinghai Key Laboratory of Animal Ecological Genomics, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Qinghai, Xining 810001, PR ChinaState Key Laboratory of Swine and Poultry Breeding Industry, Key Laboratory of Livestock and Poultry Multiomics, Ministry of Agriculture and Rural Affairs, Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, College of Animal Science and Technology, Sichuan Agricultural University, Sichuan, Chengdu 611130, PR ChinaCollege of Life Science, Sichuan Agricultural University, Sichuan, Ya’an 625014, PR ChinaKey Laboratory of Adaptation and Evolution of Plateau Biota, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Qinghai, Xining 810001, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China; Qinghai Key Laboratory of Animal Ecological Genomics, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Qinghai, Xining 810001, PR China; Corresponding authors at: Key Laboratory of Adaptation and Evolution of Plateau Biota, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, No. 23, Xinning Road, Xining, Qinghai, 810001, PR China (Q.E. Yang); College of Animal Science and Technology, Sichuan Agricultural University, No. 211, Huimin road, Wenjiang district, Chengdu 61130, PR China (G.B. Zhou).State Key Laboratory of Swine and Poultry Breeding Industry, Key Laboratory of Livestock and Poultry Multiomics, Ministry of Agriculture and Rural Affairs, Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, College of Animal Science and Technology, Sichuan Agricultural University, Sichuan, Chengdu 611130, PR China; Corresponding authors at: Key Laboratory of Adaptation and Evolution of Plateau Biota, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, No. 23, Xinning Road, Xining, Qinghai, 810001, PR China (Q.E. Yang); College of Animal Science and Technology, Sichuan Agricultural University, No. 211, Huimin road, Wenjiang district, Chengdu 61130, PR China (G.B. Zhou).Introduction: Developmental competence of oocytes matured in vitro is limited due to a lack of complete understanding of metabolism and metabolic gene expression during oocyte maturation and embryo development. Conventional metabolic analysis requires a large number of samples and is not efficiently applicable in oocytes and early embryos, thereby posing challenges in identifying key metabolites and regulating their in vitro culture system. Objectives: To enhance the developmental competence of sheep oocytes, this study aimed to identify and supplement essential metabolites that were deficient in the culture systems. Methods: The metabolic characteristics of oocytes and embryos were determined using ultrasensitive metabolomics analysis on trace samples and single-cell RNA-seq. By conducting integrated analyses of metabolites in cells (oocytes and embryos) and their developmental microenvironment (follicular fluid, oviductal fluid, and in vitro culture systems), we identified key missing metabolites in the in vitro culture systems. In order to assess the impact of these key missing metabolites on oocyte development competence, we performed in vitro culture experiments. Furthermore, omics analyses were employed to elucidate the underlying mechanisms. Results: Our findings demonstrated that betaine, carnitine and creatine were the key missing metabolites in vitro culture systems and supplementation of betaine and L-carnitine significantly improved the blastocyst formation rate (67.48% and 48.61%). Through in vitro culture experiments and omics analyses, we have discovered that L-carnitine had the potential to promote fatty acid oxidation, reduce lipid content and lipid peroxidation level, and regulate spindle morphological grade through fatty acid degradation pathway. Additionally, betaine may participate in methylation modification and osmotic pressure regulation, thereby potentially improving oocyte maturation and early embryo development in sheep. Conclusion: Together, these analyses identified key metabolites that promote ovine oocyte maturation and early embryo development, while also providing a new viewpoint to improve clinical applications such as oocyte maturation or embryo culture.http://www.sciencedirect.com/science/article/pii/S2090123224003813Sheep oocyteEarly embryoMetabolomicsTranscriptomicsL-carnitineBetaine |
| spellingShingle | Bo Pan JianPeng Qin KunLin Du LuYao Zhang GongXue Jia JiangFeng Ye QiuXia Liang QiEn Yang GuangBin Zhou Integrated ultrasensitive metabolomics and single-cell transcriptomics identify crucial regulators of sheep oocyte maturation and early embryo development in vitro Journal of Advanced Research Sheep oocyte Early embryo Metabolomics Transcriptomics L-carnitine Betaine |
| title | Integrated ultrasensitive metabolomics and single-cell transcriptomics identify crucial regulators of sheep oocyte maturation and early embryo development in vitro |
| title_full | Integrated ultrasensitive metabolomics and single-cell transcriptomics identify crucial regulators of sheep oocyte maturation and early embryo development in vitro |
| title_fullStr | Integrated ultrasensitive metabolomics and single-cell transcriptomics identify crucial regulators of sheep oocyte maturation and early embryo development in vitro |
| title_full_unstemmed | Integrated ultrasensitive metabolomics and single-cell transcriptomics identify crucial regulators of sheep oocyte maturation and early embryo development in vitro |
| title_short | Integrated ultrasensitive metabolomics and single-cell transcriptomics identify crucial regulators of sheep oocyte maturation and early embryo development in vitro |
| title_sort | integrated ultrasensitive metabolomics and single cell transcriptomics identify crucial regulators of sheep oocyte maturation and early embryo development in vitro |
| topic | Sheep oocyte Early embryo Metabolomics Transcriptomics L-carnitine Betaine |
| url | http://www.sciencedirect.com/science/article/pii/S2090123224003813 |
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