{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:46:19Z","timestamp":1760147179600,"version":"build-2065373602"},"reference-count":48,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2023,1,16]],"date-time":"2023-01-16T00:00:00Z","timestamp":1673827200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004829","name":"Sichuan Provincial Science and Technology Program","doi-asserted-by":"publisher","award":["2021YFN0121","SCCXTD-2022-13","2022NYXXS032"],"award-info":[{"award-number":["2021YFN0121","SCCXTD-2022-13","2022NYXXS032"]}],"id":[{"id":"10.13039\/501100004829","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Sichuan Beef Cattle Innovation Team of National Agricultural Industrial Technology System","award":["2021YFN0121","SCCXTD-2022-13","2022NYXXS032"],"award-info":[{"award-number":["2021YFN0121","SCCXTD-2022-13","2022NYXXS032"]}]},{"DOI":"10.13039\/501100007535","name":"Central Universities of Southwest Minzu University","doi-asserted-by":"publisher","award":["2021YFN0121","SCCXTD-2022-13","2022NYXXS032"],"award-info":[{"award-number":["2021YFN0121","SCCXTD-2022-13","2022NYXXS032"]}],"id":[{"id":"10.13039\/501100007535","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>This study aimed to investigate the spatially and temporally expressed patterns and biological characteristics of TSSK1B in male yaks and explore the potential correlation between TSSK1B and male sterility of the yak hybrid offspring (termed cattle\u2013yak). First, the coding sequence (CDS) of TSSK1B was cloned by RT-PCR, and bioinformatics analysis was conducted with relevant software. Quantitative real-time PCR (RT-qPCR) was employed to detect the expression profile of TSSK1B in various tissues of male adult yaks, the spatiotemporal expression of TSSK1B in different stages of yak testes, and the differential expression of TSSK1B between yak and cattle\u2013yak testes. The cellular localization of TSSK1B was determined by immunohistochemistry (IHC). Furthermore, the methylation status of the TSSK1B promoter region was analyzed by bisulfite-sequencing PCR (BSP). The results showed that TSSK1B was 1235 bp long, including 1104 bp of the CDS region, which encoded 367 amino acids. It was a conserved gene sharing the highest homology with Bos mutus (99.67%). In addition, the bioinformatics analysis revealed that TSSK1B was an unstable hydrophilic protein mainly containing the alpha helix of 34.06% and a random coil of 44.41%, with a transmembrane structure of 29 amino acids long. The RT-qPCR results demonstrated that TSSK1B was specifically expressed in yak testes compared with that in other tissues and especially highly expressed in adult yak testes. On the contrary, TSSK1B was hardly expressed in the testis of adult cattle\u2013yak. IHC confirmed that TSSK1B protein was more strongly expressed in the testes of adult yaks than in their fetal and juvenile counterparts. Interestingly, nearly no expression was observed in the testes of cattle\u2013yak compared with the corresponding testes of yak. Bisulfite-sequencing PCR (BSP) revealed that the methylated CpG sites in the TSSK1B promoter region of cattle\u2013yak was significantly higher than that in the yak. Taken together, this study revealed that TSSK1B was specifically expressed in yak testes and highly expressed upon sexual maturity. Moreover, the rare expression in cattle\u2013yak may be related to the hypermethylation of the promoter region, thereby providing a basis for further studies on the regulatory mechanism of TSSK1B in male cattle\u2013yak sterility.<\/jats:p>","DOI":"10.3390\/ani13020320","type":"journal-article","created":{"date-parts":[[2023,1,17]],"date-time":"2023-01-17T02:10:32Z","timestamp":1673921432000},"page":"320","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["The Biological Characteristics and Differential Expression Patterns of TSSK1B Gene in Yak and Its Infertile Hybrid Offspring"],"prefix":"10.3390","volume":"13","author":[{"given":"Yanjin","family":"Zhu","sequence":"first","affiliation":[{"name":"Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Reservation and Exploitation of Ministry of Education, Southwest Minzu University, Chengdu 610041, China"},{"name":"Key Laboratory of Animal Science of National Ethnic Affairs Commission, Southwest Minzu University, Chengdu 610041, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bangting","family":"Pan","sequence":"additional","affiliation":[{"name":"Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Reservation and Exploitation of Ministry of Education, Southwest Minzu University, Chengdu 610041, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xixi","family":"Fei","sequence":"additional","affiliation":[{"name":"Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Reservation and Exploitation of Ministry of Education, Southwest Minzu University, Chengdu 610041, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yulei","family":"Hu","sequence":"additional","affiliation":[{"name":"Key Laboratory of Animal Science of National Ethnic Affairs Commission, Southwest Minzu University, Chengdu 610041, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Manzhen","family":"Yang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Animal Science of National Ethnic Affairs Commission, Southwest Minzu University, Chengdu 610041, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hailing","family":"Yu","sequence":"additional","affiliation":[{"name":"Key Laboratory of Animal Science of National Ethnic Affairs Commission, Southwest Minzu University, Chengdu 610041, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jian","family":"Li","sequence":"additional","affiliation":[{"name":"Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Reservation and Exploitation of Ministry of Education, Southwest Minzu University, Chengdu 610041, China"},{"name":"Key Laboratory of Animal Science of National Ethnic Affairs Commission, Southwest Minzu University, Chengdu 610041, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5470-122X","authenticated-orcid":false,"given":"Xianrong","family":"Xiong","sequence":"additional","affiliation":[{"name":"Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Reservation and Exploitation of Ministry of Education, Southwest Minzu University, Chengdu 610041, China"},{"name":"Key Laboratory of Animal Science of National Ethnic Affairs Commission, Southwest Minzu University, Chengdu 610041, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,1,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"3461","DOI":"10.3390\/ani11123461","article-title":"Polymorphisms of SORBS 1 Gene and Their Correlation with Milk Fat Traits of Cattleyak","volume":"11","author":"Yang","year":"2021","journal-title":"Animals"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1016\/j.anireprosci.2018.04.067","article-title":"Isolation and characterization of spermatogenic cells from cattle, yak and cattleyak","volume":"193","author":"Shah","year":"2018","journal-title":"Anim. Reprod. Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1186\/s13568-021-01259-9","article-title":"Comparative study of growth performance, nutrient digestibility, and ruminal and fecal bacterial community between yaks and cattle-yaks raised by stall-feeding","volume":"11","author":"Dai","year":"2021","journal-title":"AMB Express"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"3927","DOI":"10.1002\/jsfa.11032","article-title":"Comparison of carcass characteristics and meat quality between Simmental crossbred cattle, cattle-yaks and Xuanhan yellow cattle","volume":"101","author":"Wang","year":"2021","journal-title":"J. Sci. Food Agric."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"358","DOI":"10.1071\/AN16111","article-title":"Fatty acid profile of ghee derived from two genotypes (cattle-yak vs yak) grazing different alpine Himalayan pasture sites","volume":"58","author":"Marquardt","year":"2016","journal-title":"Anim. Prod. Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"999","DOI":"10.1111\/rda.14166","article-title":"Disruption of cell proliferation and apoptosis balance in the testes of crossbred cattle-yaks affects spermatogenic cell fate and sterility","volume":"57","author":"Shimazaki","year":"2022","journal-title":"Reprod. Domest. Anim."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Wu, S., Mipam, T.D., Xu, C., Zhao, W., Shah, M.A., Yi, C., Luo, H., Cai, X., and Zhong, J. (2020). Testis transcriptome profiling identified genes involved in spermatogenic arrest of cattleyak. PLoS ONE, 15.","DOI":"10.1371\/journal.pone.0229503"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Zhao, W., Ahmed, S., Liu, J., Ahmed, S., Quansah, E., Solangi, T.H., Wu, Y., Yangliu, Y., Wang, H., and Zhu, J. (2021). Comparative iTRAQ proteomics identified proteins associated with sperm maturation between yak and cattleyak epididymis. BMC Vet. Res., 17.","DOI":"10.1186\/s12917-021-02907-9"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Bononi, A., Agnoletto, C., De Marchi, E., Marchi, S., Patergnani, S., Bonora, M., Giorgi, C., Missiroli, S., Poletti, F., and Rimessi, A. (2011). Protein kinases and phosphatases in the control of cell fate. Enzym. Res., 2011.","DOI":"10.4061\/2011\/329098"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1016\/0378-1119(94)90762-5","article-title":"Characterization of a novel murine testis-specific serine\/threonine kinase","volume":"139","author":"Bielke","year":"1994","journal-title":"Gene"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1851","DOI":"10.1083\/jcb.139.7.1851","article-title":"A novel family of serine\/threonine kinases participating in spermiogenesis","volume":"139","author":"Kueng","year":"1997","journal-title":"J. Cell Biol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1006\/geno.2001.6628","article-title":"Cloning and chromosomal localization of a gene encoding a novel serine\/threonine kinase belonging to the subfamily of testis-specific kinases","volume":"77","author":"Visconti","year":"2001","journal-title":"Genomics"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1093\/molehr\/gah052","article-title":"Expression analysis of the human testis-specific serine\/threonine kinase (TSSK) homologues. A TSSK member is present in the equatorial segment of human sperm","volume":"10","author":"Hao","year":"2004","journal-title":"Mol. Hum. Reprod."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"4250","DOI":"10.1128\/MCB.25.10.4250-4261.2005","article-title":"Identification and characterization of SSTK, a serine\/threonine protein kinase essential for male fertility","volume":"25","author":"Spiridonov","year":"2005","journal-title":"Mol. Cell. Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1093\/molehr\/gaq071","article-title":"Expression and localization of five members of the testis-specific serine kinase (Tssk) family in mouse and human sperm and testis","volume":"17","author":"Li","year":"2011","journal-title":"Mol. Hum. Reprod."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1111\/j.2047-2927.2012.00021.x","article-title":"Evolution of testis-specific kinases TSSK1B and TSSK2 in primates","volume":"1","author":"Shang","year":"2013","journal-title":"Andrology"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1016\/j.ydbio.2008.03.047","article-title":"Targeted deletion of Tssk1 and 2 causes male infertility due to haploinsufficiency","volume":"319","author":"Xu","year":"2008","journal-title":"Dev. Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1530\/REP-13-0224","article-title":"PPP1CC2 can form a kinase\/phosphatase complex with the testis-specific proteins TSSK1 and TSKS in the mouse testis","volume":"147","author":"MacLeod","year":"2014","journal-title":"Reproduction"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1016\/j.theriogenology.2016.09.047","article-title":"Comparative analysis of testis transcriptomes associated with male infertility in cattleyak","volume":"88","author":"Cai","year":"2017","journal-title":"Theriogenology"},{"key":"ref_20","unstructured":"Zhao, D., Wu, J., Ma, Y., Zhang, J., Feng, X., Fan, Y., Xiong, X., Fu, W., Li, J., and Xiong, Y. (2022). The molecular characteristic analysis of TRIB2 gene and its expressional patterns in Bos grunniens tissue and granulosa cells. Anim. Biotechnol., 1\u20139."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1016\/j.ijbiomac.2022.03.212","article-title":"Molecular characterization, expression and anti-tumor function analysis of yak IFITM2 gene","volume":"209","author":"Wang","year":"2022","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2380","DOI":"10.3390\/ani12182380","article-title":"PRAMEY: A Bovid-Specific Y-Chromosome Multicopy Gene Is Highly Related to Postnatal Testicular Growth in Hu Sheep","volume":"12","author":"Pei","year":"2022","journal-title":"Animals"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.theriogenology.2022.07.012","article-title":"Molecular characterization of TRIB1 gene and its role in regulation of steroidogenesis in Bos grunniens granulosa cells","volume":"191","author":"Zhao","year":"2022","journal-title":"Theriogenology"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1186\/s13072-022-00466-3","article-title":"Sperm DNA methylation alterations from cannabis extract exposure are evident in offspring","volume":"15","author":"Schrott","year":"2022","journal-title":"Epigenet. Chromatin"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1016\/j.theriogenology.2022.08.016","article-title":"DNA methylome of primary spermatocyte reveals epigenetic dysregulation associated with male sterility of cattleyak","volume":"191","author":"Luo","year":"2022","journal-title":"Theriogenology"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"14908","DOI":"10.4238\/2015.October.18.56","article-title":"Cloning, sequence characterization, and expression patterns of members of the porcine TSSK family","volume":"14","author":"Wang","year":"2015","journal-title":"Genet. Mol. Res."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Li, X., Li, Y., Song, W., Xie, D., Zhu, F., Yang, M., and Li, Y. (2021). cDNA cloning, expression and bioinformatical analysis of Tssk genes in tree shrews. Comput. Biol. Chem., 92.","DOI":"10.1016\/j.compbiolchem.2021.107474"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1080\/07924259.2016.1161667","article-title":"Characterization, expression, and functional analysis of testis-specific serine\/threonine kinase 1 (Tssk1) in the pen shell Atrina pectinata","volume":"60","author":"Li","year":"2016","journal-title":"Invertebr. Reprod. Dev."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Kim, E.J., Kim, S.J., Park, C.J., and Nam, Y.K. (2019). Characterization of testis-specific serine\/threonine kinase 1-like (TSSK1-like) gene and expression patterns in diploid and triploid Pacific abalone (Haliotis discus hannai; Gastropoda; Mollusca) males. PLoS ONE, 14.","DOI":"10.1371\/journal.pone.0226022"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"R988","DOI":"10.1016\/j.cub.2017.07.067","article-title":"Spermatogenesis","volume":"27","author":"Nishimura","year":"2017","journal-title":"Curr. Biol."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Xue, X., Zhang, L., Li, Y., Wei, H., Wu, S., Liu, T., Liu, L., Xing, Q., Wang, S., and Bao, Z. (2021). Expression of the testis-specific serine\/threonine kinases suggests their role in spermiogenesis of bay scallop argopecten irradians. Front. Physiol., 12.","DOI":"10.3389\/fphys.2021.657559"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1242\/jcs.059949","article-title":"Functional transformation of the chromatoid body in mouse spermatids requires testis-specific serine\/threonine kinases","volume":"123","author":"Shang","year":"2010","journal-title":"J. Cell Sci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1093\/biolre\/ioaa207","article-title":"Male hybrid sterility in the cattle-yak and other bovines: A review","volume":"104","author":"Niayale","year":"2021","journal-title":"Biol. Reprod."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1159\/000492516","article-title":"Interstitial deletion of 5q22. 2q23. 1 including APC and TSSK1B in a patient with adenomatous polyposis and asthenoteratozoospermia","volume":"9","author":"Kadiyska","year":"2018","journal-title":"Mol. Syndromol."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Li, J., Zhou, Y., Zhou, Z., Li, C., Wei, J., Qin, Y., Xiang, Z., Ma, H., Zhang, Y., and Zhang, Y. (2020). Comparative transcriptome analysis of three gonadal development stages reveals potential genes involved in gametogenesis of the fluted giant clam (Tridacna squamosa). BMC Genom., 21.","DOI":"10.1186\/s12864-020-07276-5"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Luo, S., Gao, X., Ding, J., Liu, C., Du, C., Hou, C., Zhu, J., and Lou, B. (2019). Transcriptome sequencing reveals the traits of spermatogenesis and testicular development in large yellow croaker (Larimichthys crocea). Genes, 10.","DOI":"10.3390\/genes10120958"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"797","DOI":"10.1042\/EBC20190037","article-title":"The DNA methylation landscape in cancer","volume":"63","author":"Skvortsova","year":"2019","journal-title":"Essays Biochem."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1186\/s13059-022-02728-5","article-title":"Large-scale manipulation of promoter DNA methylation reveals context-specific transcriptional responses and stability","volume":"23","author":"Nguyen","year":"2022","journal-title":"Genome Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"222","DOI":"10.1080\/10495398.2014.1002563","article-title":"Yak response to high-altitude hypoxic stress by altering mRNA expression and DNA methylation of hypoxia-inducible factors","volume":"26","author":"Xiong","year":"2015","journal-title":"Anim. Biotechnol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1186\/s13148-022-01324-5","article-title":"DNA methylation analysis of normal colon organoids from familial adenomatous polyposis patients reveals novel insight into colon cancer development","volume":"14","author":"Devall","year":"2022","journal-title":"Clin. Epigenet."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1038\/s41419-019-1744-5","article-title":"Roles of circular RNAs in immune regulation and autoimmune diseases","volume":"10","author":"Zhou","year":"2019","journal-title":"Cell Death Dis."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1186\/s13148-022-01337-0","article-title":"Liquid biopsies based on DNA methylation as biomarkers for the detection and prognosis of lung cancer","volume":"14","author":"Li","year":"2022","journal-title":"Clin. Epigenet."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"2485","DOI":"10.1038\/s41467-021-22720-0","article-title":"A pan-cancer analysis of CpG Island gene regulation reveals extensive plasticity within Polycomb target genes","volume":"12","author":"Zheng","year":"2021","journal-title":"Nat. Commun."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"484","DOI":"10.1038\/nrg3230","article-title":"Functions of DNA methylation: Islands, start sites, gene bodies and beyond","volume":"13","author":"Jones","year":"2012","journal-title":"Nat. Rev. Genet."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1038\/nature14192","article-title":"Function and information content of DNA methylation","volume":"517","year":"2015","journal-title":"Nature"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"811","DOI":"10.1038\/ng.864","article-title":"Dynamic CpG island methylation landscape in oocytes and preimplantation embryos","volume":"43","author":"Smallwood","year":"2011","journal-title":"Nat. Genet."},{"key":"ref_47","first-page":"23","article-title":"Underexpression of hsa-miR-449 family and their promoter hypermethylation in infertile men: A case-control study","volume":"19","author":"Najafipour","year":"2021","journal-title":"Int. J. Reprod. Biomed."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1007\/s43032-019-00025-4","article-title":"Expression and methylation pattern of hsa-miR-34 family in sperm samples of infertile men","volume":"27","author":"Momeni","year":"2020","journal-title":"Reprod. Sci."}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/13\/2\/320\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:07:48Z","timestamp":1760119668000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/13\/2\/320"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,1,16]]},"references-count":48,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2023,1]]}},"alternative-id":["ani13020320"],"URL":"https:\/\/doi.org\/10.3390\/ani13020320","relation":{},"ISSN":["2076-2615"],"issn-type":[{"type":"electronic","value":"2076-2615"}],"subject":[],"published":{"date-parts":[[2023,1,16]]}}}