{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T02:56:32Z","timestamp":1791860192739,"version":"4.3.4"},"reference-count":41,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2025,5,20]],"date-time":"2025-05-20T00:00:00Z","timestamp":1747699200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Hebei Provincial College Science and Technology Research Project","award":["QN2024004"],"award-info":[{"award-number":["QN2024004"]}]},{"name":"Hebei Provincial College Science and Technology Research Project","award":["C2023204277"],"award-info":[{"award-number":["C2023204277"]}]},{"name":"Hebei Provincial College Science and Technology Research Project","award":["226Z7603G"],"award-info":[{"award-number":["226Z7603G"]}]},{"name":"Hebei Provincial Natural Science Foundation Project","award":["QN2024004"],"award-info":[{"award-number":["QN2024004"]}]},{"name":"Hebei Provincial Natural Science Foundation Project","award":["C2023204277"],"award-info":[{"award-number":["C2023204277"]}]},{"name":"Hebei Provincial Natural Science Foundation Project","award":["226Z7603G"],"award-info":[{"award-number":["226Z7603G"]}]},{"name":"Hebei Provincial Central Guidance on Local Science and Technology Development Fund Project","award":["QN2024004"],"award-info":[{"award-number":["QN2024004"]}]},{"name":"Hebei Provincial Central Guidance on Local Science and Technology Development Fund Project","award":["C2023204277"],"award-info":[{"award-number":["C2023204277"]}]},{"name":"Hebei Provincial Central Guidance on Local Science and Technology Development Fund Project","award":["226Z7603G"],"award-info":[{"award-number":["226Z7603G"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Semen cryopreservation is a crucial technology in the artificial insemination of livestock and poultry. It not only contributes to the conservation of germplasm resources but also facilitates the cross-regional exchange of high-quality breeding stock. In this study, 165 Duroc boars were selected for genome-wide genotyping, and the sperm freezing\/thawing motility ratio (sperm recovery rate) was used as phenotypic data for genome-wide association analysis (GWAS). Considerable individual variations in sperm recovery rates (SRRs) were detected, and the sperm structure after cyropreservation was significantly better in highly freeze-tolerant individuals compared to non-freeze-tolerant ones. The heritability of the SRR was calculated and found to be 0.199 \u00b1 0.158, representing low heritability. Through GWAS, eight single-nucleotide polymorphism (SNP) loci and four candidate genes (SLC10A6, MYRF, GGA1, and UTRN) were identified as being significantly associated with sperm freezing tolerance. Moreover, the dominant genotypes of four SNPs were finally determined to be valuable for identifying individuals with high sperm freezing tolerance. This study reveals the heritability of the sperm recovery rate and identifies molecular markers associated with sperm freezing tolerance in Duroc boars, which is of great significance for accelerating boar genetic improvement and enhancing the economic efficiency of pig breeding industry.<\/jats:p>","DOI":"10.3390\/ani15101474","type":"journal-article","created":{"date-parts":[[2025,5,20]],"date-time":"2025-05-20T08:41:12Z","timestamp":1747730472000},"page":"1474","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Genome-Wide Association Study That Identifies Molecular Markers with Freezing Resistance in Duroc Boar Sperm"],"prefix":"10.3390","volume":"15","author":[{"given":"Jiajun","family":"Zhang","sequence":"first","affiliation":[{"name":"College of Animal Science and Technology, Hebei Agricultural University, Baoding 071000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meicheng","family":"Li","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Hebei Agricultural University, Baoding 071000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guangxiang","family":"Chen","sequence":"additional","affiliation":[{"name":"Experimental Training Center, Hebei Agricultural University, Baoding 071000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chenyu","family":"Tao","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Hebei Agricultural University, Baoding 071000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bushe","family":"Li","sequence":"additional","affiliation":[{"name":"Shanghai Engineering Research Center of Breeding Pig, Shanghai 201106, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hejun","family":"Zhang","sequence":"additional","affiliation":[{"name":"Shanghai Engineering Research Center of Breeding Pig, Shanghai 201106, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongyang","family":"Wang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Livestock and Poultry Resources (Pig) Evaluation and Utilization, Ministry of Agriculture and Rural Affairs, Institute of Animal Husbandry &Veterinary Science, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenjun","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Hebei Agricultural University, Baoding 071000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,5,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1016\/j.rbmo.2018.05.012","article-title":"Sperm Cryo-preservation: A Review on Current Molecular Cryobiology and Advanced Approaches","volume":"37","author":"Hezavehei","year":"2018","journal-title":"Reprod. Biomed. Online"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"106904","DOI":"10.1016\/j.anireprosci.2021.106904","article-title":"Advances in Sperm Cryopreservation in Farm Animals: Cattle, Horse, Pig and Sheep","volume":"246","author":"Yeste","year":"2022","journal-title":"Anim. Reprod. Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1111\/rda.12569","article-title":"Recent Advances in Boar Sperm Cryopreservation: State of the Art and Current Perspectives","volume":"50","author":"Yeste","year":"2015","journal-title":"Reprod. Domest. Anim."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1186\/s40813-017-0062-5","article-title":"Boar Management and Semen Handling Factors Affect the Quality of Boar Extended Semen","volume":"3","author":"Arsenakis","year":"2017","journal-title":"Porc. Health Manag."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"887","DOI":"10.1530\/rep.1.01049","article-title":"Within and between Breed Differences in Freezing Tolerance and Plasma Membrane Fatty Acid Composition of Boar Sperm","volume":"131","author":"Waterhouse","year":"2006","journal-title":"Reproduction"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1016\/j.theriogenology.2022.06.023","article-title":"Promoter Polymorphisms in STK35 and IFT27 Genes and Their Associations with Boar Sperm Freezability","volume":"189","author":"Brym","year":"2022","journal-title":"Theriogenology"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Wang, W., Bai, Y., Wang, X., Zhang, J., Li, B., Zhang, H., Zhou, X., Wang, H., and Liu, B. (2025). Identification of Key Genes and Variants Associated with Boar Sperm Freezability Using Whole Genome Resequencing. Int. J. Bio-Log. Macromol., 294.","DOI":"10.1016\/j.ijbiomac.2024.139268"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"400","DOI":"10.1016\/j.theriogenology.2019.11.001","article-title":"Transcriptome Analysis of Boar Spermatozoa with Different Freezability Using RNA-Seq","volume":"142","author":"Fraser","year":"2020","journal-title":"Theriogenology"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Gao, N., Chen, Y., Liu, X., Zhao, Y., Zhu, L., Liu, A., Jiang, W., Peng, X., Zhang, C., and Tang, Z. (2019). Weighted Single-Step GWAS Identified Candidate Genes Associated with Semen Traits in a Duroc Boar Population. BMC Genom., 20.","DOI":"10.1186\/s12864-019-6164-5"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"skab188","DOI":"10.1093\/jas\/skab188","article-title":"Identification of new semen trait-related candidate genes in Duroc boars through genome-wide association and weighted gene co-expression network analyses","volume":"99","author":"Mei","year":"2021","journal-title":"J. Anim. Sci."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Zhuang, Z., Li, K., Yang, K., Gao, G., Li, Z., Zhu, X., and Zhao, Y. (2024). Genome-Wide Association Study Reveals Novel Candidate Genes Influencing Semen Traits in Landrace Pigs. Animals, 14.","DOI":"10.3390\/ani14131839"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/j.meatsci.2015.04.011","article-title":"The Inclusion of Duroc Breed in Maternal Line Affects Pork Quality and Fatty Acid Profile","volume":"107","author":"Alonso","year":"2015","journal-title":"Meat Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"117395","DOI":"10.1016\/j.theriogenology.2025.117395","article-title":"Sperm Metabolomics Identifies Freezability Markers in Duroc, Landrace, and Large White Boars","volume":"240","author":"Li","year":"2025","journal-title":"Theriogenology"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Liu, J., Zeng, Y., Tian, Y., Cheng, L., Li, W., Cheng, S., Wang, J., and Li, L. (2025). Genome-Wide Association Study Reveals Novel Loci and Candidate Genes for Birth Weight in Pigs. Animals, 15.","DOI":"10.3390\/ani15060825"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Zeng, H., Zhong, Z., Xu, Z., Teng, J., Wei, C., Chen, Z., Zhang, W., Ding, X., Li, J., and Zhang, Z. (2022). Meta-analysis of ge-nome-wide association studies uncovers shared candidate genes across breeds for pig fatness trait. BMC Genom., 23.","DOI":"10.1186\/s12864-022-09036-z"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Gualtieri, R., Kalthur, G., Barbato, V., Di Nardo, M., Adiga, S.K., and Talevi, R. (2021). Mitochondrial Dysfunction and Oxidative Stress Caused by Cryopreservation in Reproductive Cells. Antioxidants, 10.","DOI":"10.3390\/antiox10030337"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1038\/s41420-024-02013-3","article-title":"Sperm Freezing Damage: The Role of Regulated Cell Death","volume":"10","author":"Hai","year":"2024","journal-title":"Cell Death Discov."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"697","DOI":"10.1146\/annurev.cellbio.19.111301.140101","article-title":"Plasma Membrane Disruption: Repair, Prevention, Adaptation","volume":"19","author":"McNeil","year":"2003","journal-title":"Annu. Rev. Cell Dev. Biol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1700","DOI":"10.1016\/j.theriogenology.2012.05.002","article-title":"Effects of Reactive Oxygen Species on Sperm Function","volume":"78","author":"Guthrie","year":"2012","journal-title":"Theriogenology"},{"key":"ref_20","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. Reprod."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"105436","DOI":"10.1016\/j.livsci.2024.105436","article-title":"Estimation of Genetic Parameters and Breeding Value Accuracy for Growth and Egg Production Traits in Korean Native Chicken Pure Lines","volume":"282","author":"Afrin","year":"2024","journal-title":"Livest. Sci."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1038\/nrg2322","article-title":"Heritability in the Genomics Era\u2014Concepts and Misconceptions","volume":"9","author":"Visscher","year":"2008","journal-title":"Nat. Rev. Genet."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"202","DOI":"10.1016\/j.theriogenology.2018.10.006","article-title":"Genetics of Bull Semen Characteristics in a Multi-Breed Cattle Popu-lation","volume":"123","author":"Berry","year":"2019","journal-title":"Theriogenology"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1016\/S0378-4320(03)00142-8","article-title":"Differences among Breeds and Manifestation of Heterosis in AI Boar Sperm Output","volume":"80","author":"Smital","year":"2004","journal-title":"Anim. Reprod. Sci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1177","DOI":"10.1111\/rda.13492","article-title":"Genetic Parameters for Semen Production Traits in Swiss Dairy Bulls","volume":"54","author":"Burren","year":"2019","journal-title":"Reprod. Domest. Anim."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Fietz, D., Bakhaus, K., Wapelhorst, B., Grosser, G., G\u00fcnther, S., Alber, J., D\u00f6ring, B., Kliesch, S., Weidner, W., and Galuska, C.E. (2013). Membrane Transporters for Sulfated Steroids in the Human Testis\u2014Cellular Localization, Expression Pattern and Functional Analysis. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0062638"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1016\/j.anireprosci.2011.02.008","article-title":"Time Trends, Environmental Factors and Genetic Basis of Semen Traits Collected in Holstein Bulls under Commercial Conditions","volume":"124","author":"Karoui","year":"2011","journal-title":"Anim. Reprod. Sci."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1016\/S0921-4488(00)00140-1","article-title":"Reproductive Characteristics of Ethiopian Highland Sheep","volume":"37","author":"Toe","year":"2000","journal-title":"Small Rumin. Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1111\/j.1439-0388.2008.00788.x","article-title":"Estimation of Genetic Parameters and Genome Scan for 15 Semen Characteristics Traits of Holstein Bulls","volume":"126","author":"Druet","year":"2009","journal-title":"J. Anim. Breed. Genet."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Wannowius, M., Karakus, E., Akt\u00fcrk, Z., Breuer, J., and Geyer, J. (2023). Role of the Sodium-Dependent Organic Anion Transporter (SOAT\/SLC10A6) in Physiology and Pathophysiology. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24129926"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/j.jsbmb.2017.07.019","article-title":"Sodium-Dependent Organic Anion Transporter (Slc10a6\u2212\/\u2212) Knockout Mice Show Normal Spermatogenesis and Reproduction, but Elevated Serum Levels for Cholesterol Sulfate","volume":"179","author":"Bakhaus","year":"2018","journal-title":"J. Steroid Biochem. Mol. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"19728","DOI":"10.1074\/jbc.M702663200","article-title":"Cloning and Functional Characterization of Human Sodium-Dependent Organic Anion Transporter (SLC10A6)","volume":"282","author":"Geyer","year":"2007","journal-title":"J. Bio-Log. Chem"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1016\/j.jsbmb.2013.03.009","article-title":"Cloning and Functional Characterization of the Mouse Sodium-Dependent Organic Anion Transporter Soat (Slc10a6)","volume":"138","author":"Grosser","year":"2013","journal-title":"J. Steroid Biochem. Mol. Biol."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Bujalka, H., Koenning, M., Jackson, S., Perreau, V.M., Pope, B., Hay, C.M., Mitew, S., Hill, A.F., Lu, Q.R., and Wegner, M. (2013). MYRF Is a Membrane-Associated Transcription Factor That Autoproteolytically Cleaves to Directly Activate Myelin Genes. PLoS Biol., 11.","DOI":"10.1371\/journal.pbio.1001625"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2319","DOI":"10.1093\/hmg\/ddz066","article-title":"MYRF haploinsufficiency causes 46,XY and 46,XX disorders of sex development: Bioinformatics consideration","volume":"28","author":"Hamanaka","year":"2019","journal-title":"Hum. Mol. Genet."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"e15673","DOI":"10.7717\/peerj.15673","article-title":"GGA1 Participates in Spermatogenesis in Mice under Stress","volume":"11","author":"Jiao","year":"2023","journal-title":"PeerJ"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Hirst, J., and Carmichael, J. (2011). A Potential Role for the Clathrin Adaptor GGA in Drosophila Spermatogenesis. BMC Cell Biol., 12.","DOI":"10.1186\/1471-2121-12-22"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Khawar, M.B., Gao, H., and Li, W. (2019). Mechanism of Acrosome Biogenesis in Mammals. Front. Cell Dev. Biol, 7.","DOI":"10.3389\/fcell.2019.00195"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Venditti, M., and Minucci, S. (2022). Differential Expression and Localization of EHBP1L1 during the First Wave of Rat Spermatogenesis Suggest Its Involvement in Acrosome Biogenesis. Biomedicines, 10.","DOI":"10.3390\/biomedicines10010181"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"5899","DOI":"10.1074\/jbc.M709410200","article-title":"Generation and Characterization of Transgenic Mice with the Full-Length Human DMD Gene","volume":"283","author":"Boer","year":"2008","journal-title":"J. Biol. Chem."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Chen, H.-C., Chin, Y.-F., Lundy, D.J., Liang, C.-T., Chi, Y.-H., Kuo, P., and Hsieh, P.C.H. (2017). Utrophin Compensates Dys-trophin Loss during Mouse Spermatogenesis. Sci. Rep., 7.","DOI":"10.1038\/s41598-017-05993-8"}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/15\/10\/1474\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:35:46Z","timestamp":1760031346000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/15\/10\/1474"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,5,20]]},"references-count":41,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2025,5]]}},"alternative-id":["ani15101474"],"URL":"https:\/\/doi.org\/10.3390\/ani15101474","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,5,20]]}}}