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Our prior study demonstrated that mitochondrial regulators, SIRT5 and IDH2, were significantly downregulated during cryopreservation; however, how this dysregulation alters sperm metabolism remains unknown. Here, SIRT5 and IDH2 were knocked down in boar sperm via siRNA electro-transfection, and differential metabolites were identified by liquid chromatography\u2013tandem mass spectrometry (LC-MS\/MS). Results showed that SIRT5\/IDH2 knockdown dysregulated antioxidant pathways and significantly downregulated L-methionine (p\u2009&amp;lt;\u20090.05). Supplementation with 80\u2009\u00b5M L-methionine significantly increased total sperm motility (fresh: \u223c+6% at 48\u2009hr; cryopreserved: \u223c+10%), progressive motility, ATP content, and mitochondrial function, while maintaining acrosomal and plasma membrane integrity. Notably, L-methionine reduced reactive oxygen species levels by 9% (p\u2009&amp;lt;\u20090.05) in fresh sperm at 48\u2009hr and 13% in post-thawed sperm. This study elucidates a mechanism-driven strategy wherein the SIRT5\/IDH2 axis maintains sperm antioxidant capacity by regulating L-methionine and associated metabolites. These findings enhance our understanding of energy metabolism in sperm and offer a targeted approach to improve cryopreserved semen quality, potentially increasing artificial insemination success rates.<\/jats:p>","DOI":"10.1093\/reprod\/xaag017","type":"journal-article","created":{"date-parts":[[2026,1,21]],"date-time":"2026-01-21T12:40:16Z","timestamp":1768999216000},"source":"Crossref","is-referenced-by-count":3,"title":["L-Methionine enhances cryosurvival and antioxidant activity of boar sperm"],"prefix":"10.1093","volume":"171","author":[{"given":"Malik Ahsan","family":"Ali","sequence":"first","affiliation":[{"name":"State Key Laboratory of Swine and Poultry Breeding Industry, College of Animal Science and Technology, Sichuan Agricultural University , Chengdu,","place":["China"]},{"name":"Key Laboratory of Livestock and Poultry Multi-Omics, Ministry of Agriculture and Rural Affairs, College of Animal Science and Technology, Sichuan Agricultural University , 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Agricultural University , Chengdu,","place":["China"]},{"name":"Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, College of Animal Science and Technology, Sichuan Agricultural University , Chengdu,","place":["China"]},{"name":"College of Animal Science and Technology, Sichuan Agricultural University , Chengdu,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weihan","family":"Yang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Swine and Poultry Breeding Industry, College of Animal Science and Technology, Sichuan Agricultural University , Chengdu,","place":["China"]},{"name":"Key Laboratory of Livestock and Poultry Multi-Omics, Ministry of Agriculture and Rural Affairs, College of Animal Science and Technology, Sichuan Agricultural University , Chengdu,","place":["China"]},{"name":"Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, College of Animal Science and Technology, Sichuan Agricultural University , Chengdu,","place":["China"]},{"name":"College of Animal Science and Technology, Sichuan Agricultural University , Chengdu,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qingyong","family":"Ni","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Swine and Poultry Breeding Industry, College of Animal Science and Technology, Sichuan Agricultural University , Chengdu,","place":["China"]},{"name":"Key Laboratory of Livestock and Poultry Multi-Omics, Ministry of Agriculture and Rural Affairs, College of Animal Science and Technology, Sichuan Agricultural University , Chengdu,","place":["China"]},{"name":"Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, College of Animal Science and Technology, Sichuan Agricultural University , Chengdu,","place":["China"]},{"name":"College of Animal Science and Technology, Sichuan Agricultural 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