{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,10]],"date-time":"2025-12-10T05:25:12Z","timestamp":1765344312877,"version":"3.46.0"},"reference-count":47,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2025,12,8]],"date-time":"2025-12-08T00:00:00Z","timestamp":1765152000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Biological Breeding-National Science and Technology Major Project","award":["2023ZD0404405-03"],"award-info":[{"award-number":["2023ZD0404405-03"]}]},{"DOI":"10.13039\/501100009997","name":"earmarked fund for Modern Agro-industry Technology Research System","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100009997","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Sperm cryo-tolerance resulted in significant variations in post-thaw semen quality among breeds and individual boars. In the present study, semen samples from thirty-seven large white boars were cryopreserved to select individuals with strong and weak freezing tolerance according to their post-thaw sperm quality. Comparative TMT-based quantitative proteomic analysis between the two groups identified 22 significantly differentially expressed proteins. NT5C1B and ADA, the significantly downregulated proteins in the semen of the low cryo-tolerance group, were supplemented in the semen samples with lower cryo-tolerance. Supplementation with 1 \u00b5g\/mL of NT5C1B dramatically (p &lt; 0.05) improved kinematic parameters and structural integrity. In comparison with the control group, mitochondrial activity and antioxidant capacity were significantly enhanced in post-thaw sperm. In vitro fertilization assays revealed that the NT5C1B-treated group also has notably (p &lt; 0.05) high sperm penetration and embryonic cleavage rates. ADA supplementation did not exhibit obvious freezing tolerance effects. NT5C1B can be a potential key functional protein to enhance the quality and cryo-tolerance during cryopreservation. Specifically, supplementation with 1 \u00b5g\/mL NT5C1B significantly improved post-thaw motility, structural integrity, mitochondrial activity, and antioxidant capacity and ultimately enhanced the sperm penetration rate and embryonic cleavage rate in cryo-sensitive sperm, confirming its role as a functional protector during cryopreservation.<\/jats:p>","DOI":"10.3390\/ani15243530","type":"journal-article","created":{"date-parts":[[2025,12,8]],"date-time":"2025-12-08T09:17:36Z","timestamp":1765185456000},"page":"3530","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["NT5C1B Improves Fertility of Boar Spermatozoa by Enhancing Quality and Cryotolerance During Cryopreservation"],"prefix":"10.3390","volume":"15","author":[{"given":"Shibin","family":"Wang","sequence":"first","affiliation":[{"name":"College of Animal Science, Shanxi Agricultural University, Jinzhong 030801, China"},{"name":"Shanxi Key Laboratory of Animal Genetics Resource Utilization and Breeding, Jinzhong 030801, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lei","family":"Shi","sequence":"additional","affiliation":[{"name":"College of Animal Science, Shanxi Agricultural University, Jinzhong 030801, China"},{"name":"Shanxi Key Laboratory of Animal Genetics Resource Utilization and Breeding, Jinzhong 030801, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhaoyang","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Animal Science, Shanxi Agricultural University, Jinzhong 030801, China"},{"name":"Shanxi Key Laboratory of Animal Genetics Resource Utilization and Breeding, Jinzhong 030801, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Junjie","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Animal Science, Shanxi Agricultural University, Jinzhong 030801, China"},{"name":"Shanxi Key Laboratory of Animal Genetics Resource Utilization and Breeding, Jinzhong 030801, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jiandong","family":"Xing","sequence":"additional","affiliation":[{"name":"College of Animal Science, Shanxi Agricultural University, Jinzhong 030801, China"},{"name":"Shanxi Key Laboratory of Animal Genetics Resource Utilization and Breeding, Jinzhong 030801, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jingxian","family":"Yang","sequence":"additional","affiliation":[{"name":"College of Animal Science, Shanxi Agricultural University, Jinzhong 030801, China"},{"name":"Shanxi Key Laboratory of Animal Genetics Resource Utilization and Breeding, Jinzhong 030801, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jiaxin","family":"Duan","sequence":"additional","affiliation":[{"name":"College of Animal Science, Shanxi Agricultural University, Jinzhong 030801, China"},{"name":"Shanxi Key Laboratory of Animal Genetics Resource Utilization and Breeding, Jinzhong 030801, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3844-3726","authenticated-orcid":false,"given":"Bugao","family":"Li","sequence":"additional","affiliation":[{"name":"College of Animal Science, Shanxi Agricultural University, Jinzhong 030801, China"},{"name":"Shanxi Key Laboratory of Animal Genetics Resource Utilization and Breeding, Jinzhong 030801, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Guoqing","family":"Cao","sequence":"additional","affiliation":[{"name":"College of Animal Science, Shanxi Agricultural University, Jinzhong 030801, China"},{"name":"Shanxi Key Laboratory of Animal Genetics Resource Utilization and Breeding, Jinzhong 030801, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e20200219","DOI":"10.1590\/1984-3143-ar2020-0219","article-title":"The effects of cryopreservation on the acrosome structure, enzyme activity, motility, and fertility of bovine, ovine, and goat sperm","volume":"17","author":"Sun","year":"2021","journal-title":"Anim. 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