{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,26]],"date-time":"2026-01-26T21:37:09Z","timestamp":1769463429345,"version":"3.49.0"},"reference-count":62,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2026,1,22]],"date-time":"2026-01-22T00:00:00Z","timestamp":1769040000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ningxia Key R&D Talent Recruitment Program","award":["2023BSB03064"],"award-info":[{"award-number":["2023BSB03064"]}]},{"name":"Ningxia Higher Education Natural Science Project","award":["NYC2024058"],"award-info":[{"award-number":["NYC2024058"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>In livestock farming, the reproductive function and breeding performance of Tan sheep are crucial for enhancing farming efficiency. Despite advances in research on sheep germ cells, studies on the identification of markers for spermatogonia, spermatocytes, and spermatozoa in Tan sheep remain limited and inadequate. In this study, Tan sheep were used as research subjects to investigate the morphological characteristics of testicular tissues, the developmental status of germ cells, and potential novel markers for spermatogonia, spermatocytes, and spermatozoa across different ages (0 days, 60 days, 180 days, and 365 days). Homology of the SMC3, G3BP1, and AKAP4 genes was analyzed via NCBI alignment. The localization and expression characteristics of these genes in the testis tissues of Tan sheep were investigated using HE staining, qPCR, and immunofluorescence double staining. The results showed that from 0 to 365 days of age, with increasing age, spermatogonia, spermatocytes, and spermatids exhibited an orderly distribution, and mature spermatozoa appeared in the tubular lumen, marking the initial establishment of the spermatogenic process. The homology of SMC3, G3BP1, and AKAP4 was 90%, 85%, and 81%. The mRNA levels of SMC3 and G3BP1 in the testes of 60-day-old Tan sheep were significantly increased, while AKAP4 expression showed a gradual increase with advancing age. SMC3 was co-localized with PLZF in undifferentiated spermatogonia, G3BP1 was co-expressed with SYCP2 in spermatocytes, and AKAP4 was co-expressed with PNA in spermatozoa. The findings of this study provide further supportive evidence for novel markers of spermatogonia, spermatocytes, and spermatozoa in Tan sheep.<\/jats:p>","DOI":"10.3390\/ani16020350","type":"journal-article","created":{"date-parts":[[2026,1,23]],"date-time":"2026-01-23T14:58:40Z","timestamp":1769180320000},"page":"350","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Exploration of Novel Markers in Tan Sheep Spermatogenesis"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-6724-7216","authenticated-orcid":false,"given":"Yuan","family":"Ma","sequence":"first","affiliation":[{"name":"College of Animal Science and Technology, Ningxia University, Yinchuan 750021, China"},{"name":"Key Laboratory of Ruminant Molecular Cell Breeding, Yinchuan 750021, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-2305-7767","authenticated-orcid":false,"given":"Haoyan","family":"Jin","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Ningxia University, Yinchuan 750021, China"},{"name":"Key Laboratory of Ruminant Molecular Cell Breeding, Yinchuan 750021, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-7665-9842","authenticated-orcid":false,"given":"Nana","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Ningxia University, Yinchuan 750021, China"},{"name":"Key Laboratory of Ruminant Molecular Cell Breeding, Yinchuan 750021, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-3075-1030","authenticated-orcid":false,"given":"Yaru","family":"Xie","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Ningxia University, Yinchuan 750021, China"},{"name":"Key Laboratory of Ruminant Molecular Cell Breeding, Yinchuan 750021, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-3524-4414","authenticated-orcid":false,"given":"Lingkai","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Ningxia University, Yinchuan 750021, China"},{"name":"Key Laboratory of Ruminant Molecular Cell Breeding, Yinchuan 750021, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8647-3057","authenticated-orcid":false,"given":"Bei","family":"Cai","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Ningxia University, Yinchuan 750021, China"},{"name":"Key Laboratory of Ruminant Molecular Cell Breeding, Yinchuan 750021, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,1,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2116","DOI":"10.1080\/23802359.2019.1623114","article-title":"Characterization of the complete mitochondrial genome sequence of Jialuo sheep (Ovis aries)","volume":"4","author":"Fu","year":"2019","journal-title":"Mitochondrial DNA B Resour."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1159","DOI":"10.1080\/21655979.2020.1834740","article-title":"Nutritive value and ruminal degradation of seven Chinese herbs as forage for Tan sheep","volume":"11","author":"Jiang","year":"2020","journal-title":"Bioengineered"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1186\/s40104-021-00632-9","article-title":"Dynamic changes of genomic methylation profiles at different growth stages in Chinese Tan sheep","volume":"12","author":"Liu","year":"2021","journal-title":"J. Anim. Sci. Biotechnol."},{"key":"ref_4","unstructured":"Zhao, Y. (2011). Sheep Production, China Agricultural Press. [3rd ed.]."},{"key":"ref_5","unstructured":"Yang, L. (2015). Animal Reproduction, China Agricultural Press. [4th ed.]."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"776","DOI":"10.1002\/j.1939-4640.2000.tb03408.x","article-title":"All you wanted to know about spermatogonia but were afraid to ask","volume":"21","author":"Russell","year":"2000","journal-title":"J. Androl."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/j.scr.2018.04.009","article-title":"Spermatogonial stem cells and spermatogenesis in mice, monkeys and men","volume":"29","author":"Fayomi","year":"2018","journal-title":"Stem Cell Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1141","DOI":"10.1038\/s41422-018-0099-2","article-title":"The adult human testis transcriptional cell atlas","volume":"28","author":"Guo","year":"2018","journal-title":"Cell Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2091","DOI":"10.1016\/j.theriogenology.2006.03.039","article-title":"Isolation and transplantation of spermatogonia in sheep","volume":"66","author":"Dobson","year":"2006","journal-title":"Theriogenology"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1071\/RD09237","article-title":"Expression of promyelocytic leukaemia zinc-finger in ovine testis and its application in evaluating the enrichment efficiency of differential plating","volume":"22","author":"Borjigin","year":"2010","journal-title":"Reprod. Fertil. Dev."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1016\/j.anireprosci.2013.02.007","article-title":"Fertility of cryopreserved ovine semen is determined by sperm velocity","volume":"138","author":"Bisbal","year":"2013","journal-title":"Anim. Reprod. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"e21187","DOI":"10.1096\/fj.202001035RRR","article-title":"Characterization of sheep spermatogenesis through single-cell RNA sequencing","volume":"35","author":"Yang","year":"2021","journal-title":"FASEB J."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"e22348","DOI":"10.1096\/fj.202200152R","article-title":"Single-cell RNA sequencing of the Mongolia sheep testis reveals a conserved and divergent transcriptome landscape of mammalian spermatogenesis","volume":"36","author":"Tian","year":"2022","journal-title":"FASEB J."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e23367","DOI":"10.1002\/dvg.23367","article-title":"Generation of B6-Ddx4em1(CreERT2)Utr, a novel CreERT2 knock-in line, for germ cell lineage by CRISPR\/Cas9","volume":"58","author":"Le","year":"2020","journal-title":"Genesis"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Wang, M., Ding, H., Wu, S., Wang, M., Wei, C., Wang, B., Bao, Z., and Hu, J. (2022). Vasa Is a Potential Germ Cell Marker in Leopard Coral Grouper (Plectropomus leopardus). Genes, 13.","DOI":"10.3390\/genes13061077"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"7191","DOI":"10.1093\/nar\/gkx328","article-title":"In mammalian foetal testes, SOX9 regulates expression of its target genes by binding to genomic regions with conserved signatures","volume":"45","author":"Rahmoun","year":"2017","journal-title":"Nucleic Acids Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1016\/j.theriogenology.2019.07.008","article-title":"Pig SOX9: Expression profiles of Sertoli cell (SCs) and a functional 18 bp indel affecting testis weight","volume":"138","author":"Chen","year":"2019","journal-title":"Theriogenology"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.anireprosci.2017.01.002","article-title":"Characterization of male germ cell markers in canine testis","volume":"182","author":"Lee","year":"2017","journal-title":"Anim. Reprod. Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1213","DOI":"10.1038\/s41594-025-01509-5","article-title":"ZBTB16\/PLZF regulates juvenile spermatogonial stem cell development through an extensive transcription factor poising network","volume":"32","author":"Yi","year":"2025","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"e13347","DOI":"10.1111\/cpr.13347","article-title":"Ubiquitin C-terminal hydrolase L1 (UCHL1), a double-edged sword in mammalian oocyte maturation and spermatogenesis","volume":"56","author":"Yang","year":"2023","journal-title":"Cell Prolif."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Takemoto, K., Imai, Y., Saito, K., Kawasaki, T., Carlton, P.M., Ishiguro, K.I., and Sakai, N. (2020). Sycp2 is essential for synaptonemal complex assembly, early meiotic recombination and homologous pairing in zebrafish spermatocytes. PLoS Genet., 16.","DOI":"10.1371\/journal.pgen.1008640"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1568","DOI":"10.1038\/s41467-024-45693-2","article-title":"Meiotic protein SYCP2 confers resistance to DNA-damaging agents through R-loop-mediated DNA repair","volume":"15","author":"Wang","year":"2024","journal-title":"Nat. Commun."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1007\/s00418-016-1478-8","article-title":"Identification of sperm equatorial segment protein 1 in the acrosome as the primary binding target of peanut agglutinin (PNA) in the mouse testis","volume":"147","author":"Nakata","year":"2017","journal-title":"Histochem. Cell Biol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"464","DOI":"10.1002\/mrd.22982","article-title":"Peanut agglutinin specifically binds to a sperm region between the nucleus and mitochondria in tunicates and sea urchins","volume":"85","author":"Nakazawa","year":"2018","journal-title":"Mol. Reprod. Dev."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1016\/j.exphem.2018.11.008","article-title":"Smc3 is required for mouse embryonic and adult hematopoiesis","volume":"70","author":"Wang","year":"2019","journal-title":"Exp. Hematol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"dev199800","DOI":"10.1242\/dev.199800","article-title":"Maternal Smc3 protects the integrity of the zygotic genome through DNA replication and mitosis","volume":"148","author":"Yueh","year":"2021","journal-title":"Development"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1038\/s41590-018-0262-4","article-title":"G3BP1 promotes DNA binding and activation of cGAS","volume":"20","author":"Liu","year":"2019","journal-title":"Nat. Immunol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1016\/j.ydbio.2019.06.017","article-title":"Proteomics and single-cell RNA analysis of Akap4-knockout mice model confirm indispensable role of Akap4 in spermatogenesis","volume":"454","author":"Fang","year":"2019","journal-title":"Dev. Biol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1093\/hmg\/ddab234","article-title":"Loss-of-function missense variant of AKAP4 induced male infertility through reduced interaction with QRICH2 during sperm flagella development","volume":"31","author":"Zhang","year":"2021","journal-title":"Hum. Mol. Genet."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1033","DOI":"10.2116\/analsci.20P361","article-title":"DNA Extraction with TRIzol Reagent Using a Silica Column","volume":"37","author":"Yang","year":"2021","journal-title":"Anal. Sci."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Elekwachi, C.O., Wang, Z., Wu, X., Rabee, A., and Forster, R.J. (2017). Total rRNA-Seq Analysis Gives Insight into Bacterial, Fungal, Protozoal and Archaeal Communities in the Rumen Using an Optimized RNA Isolation Method. Front. Microbiol., 8.","DOI":"10.3389\/fmicb.2017.01814"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1650","DOI":"10.1016\/j.celrep.2018.10.026","article-title":"The Mammalian Spermatogenesis Single-Cell Transcriptome, from Spermatogonial Stem Cells to Spermatids","volume":"25","author":"Hermann","year":"2018","journal-title":"Cell Rep."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"483","DOI":"10.3390\/cimb44020033","article-title":"The Transcriptional Cell Atlas of Testis Development in Sheep at Pre-Sexual Maturity","volume":"44","author":"Wu","year":"2022","journal-title":"Curr. Issues Mol. Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1159\/000439452","article-title":"Variation of Meiotic Recombination Rates and MLH1 Foci Distribution in Spermatocytes of Cattle, Sheep and Goats","volume":"146","author":"Vozdova","year":"2015","journal-title":"Cytogenet. Genome Res."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Yang, H., Deng, M., Lv, W., Wei, Z., Cai, Y., Cheng, P., Wang, F., and Zhang, Y. (2022). Overexpression of bmp4, dazl, nanos3 and sycp2 in Hu Sheep Leydig Cells Using CRISPR\/dcas9 System Promoted Male Germ Cell Related Gene Expression. Biology, 11.","DOI":"10.3390\/biology11020289"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"7860","DOI":"10.1038\/s41467-022-35465-1","article-title":"SOX17-positive rete testis epithelium is required for Sertoli valve formation and normal spermiogenesis in the male mouse","volume":"13","author":"Uchida","year":"2022","journal-title":"Nat. Commun."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"5514","DOI":"10.1093\/nar\/gkae178","article-title":"Two redundant transcription factor binding sites in a single enhancer are essential for mammalian sex determination","volume":"52","author":"Ridnik","year":"2024","journal-title":"Nucleic Acids Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1038\/s42003-025-07504-2","article-title":"GATA4 binding to the Sox9 enhancer mXYSRa\/Enh13 is critical for testis differentiation in mouse","volume":"8","author":"Ogawa","year":"2025","journal-title":"Commun. Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"841","DOI":"10.1101\/gad.14.7.841","article-title":"The mouse homolog of Drosophila Vasa is required for the development of male germ cells","volume":"14","author":"Tanaka","year":"2000","journal-title":"Genes Dev."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"e13390","DOI":"10.1111\/cpr.13390","article-title":"MAST4 controls cell cycle in spermatogonial stem cells","volume":"56","author":"Lee","year":"2023","journal-title":"Cell Prolif."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"3076","DOI":"10.1182\/bloodadvances.2023011411","article-title":"Yin Yang 1 regulates cohesin complex protein SMC3 in mouse hematopoietic stem cells","volume":"8","author":"Lu","year":"2024","journal-title":"Blood Adv."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1826","DOI":"10.1038\/s12276-024-01293-0","article-title":"SMC3 contributes to heart development by regulating super-enhancer associated genes","volume":"56","author":"Zhang","year":"2024","journal-title":"Exp. Mol. Med."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Zhang, D., Jin, W., Cui, Y., and He, Z. (2024). Establishment and Characterization of Testis Organoids with Proliferation and Differentiation of Spermatogonial Stem Cells into Spermatocytes and Spermatids. Cells, 13.","DOI":"10.3390\/cells13191642"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1016\/j.cell.2020.03.046","article-title":"G3BP1 Is a Tunable Switch that Triggers Phase Separation to Assemble Stress Granules","volume":"181","author":"Yang","year":"2020","journal-title":"Cell"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"352","DOI":"10.1016\/j.cub.2023.12.018","article-title":"Multiple reorganizations of the lateral elements of the synaptonemal complex facilitate homolog segregation in Bombyx mori oocytes","volume":"34","author":"Xiang","year":"2024","journal-title":"Curr. Biol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"102063","DOI":"10.1016\/j.molmet.2024.102063","article-title":"Essential role of germ cell glycerol-3-phosphate phosphatase for sperm health, oxidative stress control and male fertility in mice","volume":"90","author":"Oppong","year":"2024","journal-title":"Mol. Metab."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"927","DOI":"10.1016\/j.jgg.2024.12.010","article-title":"The testis-specific gene 1700030J22Rikis essential for sperm flagellar function and male fertility in mice","volume":"52","author":"Yun","year":"2025","journal-title":"J. Genet. Genom."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"689","DOI":"10.1016\/j.molcel.2010.05.027","article-title":"An Smc3 acetylation cycle is essential for establishment of sister chromatid cohesion","volume":"39","author":"Hu","year":"2010","journal-title":"Mol. Cell"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"0091","DOI":"10.34133\/research.0091","article-title":"SERBP1 Promotes Stress Granule Clearance by Regulating 26S Proteasome Activity and G3BP1 Ubiquitination and Protects Male Germ Cells from Thermostimuli Damage","volume":"6","author":"Wang","year":"2023","journal-title":"Research"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"538","DOI":"10.1007\/s13238-016-0354-6","article-title":"Synaptonemal complex protein 2 (SYCP2) mediates the association of the centromere with the synaptonemal complex","volume":"8","author":"Feng","year":"2017","journal-title":"Protein Cell."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"eadk8152","DOI":"10.1126\/sciadv.adk8152","article-title":"G3BP1-dependent condensation of translationally inactive viral RNAs antagonizes infection","volume":"10","author":"Burke","year":"2024","journal-title":"Sci. Adv."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1315","DOI":"10.2741\/3791","article-title":"Role of a-kinase anchoring proteins (AKAPs) in reproduction","volume":"16","author":"Luconi","year":"2011","journal-title":"Front. Biosci. (Landmark Ed.)"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1753","DOI":"10.1080\/15548627.2020.1783822","article-title":"Autophagy core protein ATG5 is required for elongating spermatid development, sperm individualization and normal fertility in male mice","volume":"17","author":"Huang","year":"2021","journal-title":"Autophagy"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"e12480","DOI":"10.1002\/rmb2.12480","article-title":"Influence of sperm cryopreservation on sperm motility and proAKAP4 concentration in mice","volume":"21","author":"Boersma","year":"2022","journal-title":"Reprod. Med. Biol."},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Dordas-Perpiny\u00e0, M., Yanez-Ortiz, I., Sergeant, N., Mevel, V., Bruyas, J.F., Catal\u00e1n, J., Delehedde, M., Briand-Amirat, L., and Mir\u00f3, J. (2022). ProAKAP4 Concentration Is Related to Sperm Motility and Motile Sperm Subpopulations in Frozen-Thawed Horse Semen. Animals, 12.","DOI":"10.3390\/ani12233417"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"8367714","DOI":"10.1155\/vmi\/8367714","article-title":"A-Kinase Anchor Protein 4 (proAKAP4): Protein Molecule-Based Fertility Marker of Indonesian Dairy Bull and Its Correlation with Frozen-Thawed Sperm Quality","volume":"2025","author":"Pardede","year":"2025","journal-title":"Vet. Med. Int."},{"key":"ref_57","doi-asserted-by":"crossref","unstructured":"Riesco, M.F., Anel-Lopez, L., Neila-Montero, M., Palacin-Martinez, C., Montes-Garrido, R., Alvarez, M., de Paz, P., and Anel, L. (2020). ProAKAP4 as Novel Molecular Marker of Sperm Quality in Ram: An Integrative Study in Fresh, Cooled and Cryopreserved Sperm. Biomolecules, 10.","DOI":"10.3390\/biom10071046"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"100455","DOI":"10.1016\/j.vas.2025.100455","article-title":"ProAKAP4 concentration in fresh canine semen and its correlation with motility parameters","volume":"28","author":"Bencharif","year":"2025","journal-title":"Vet. Anim. Sci."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"25","DOI":"10.4103\/aja202329","article-title":"Decreased AKAP4\/PKA signaling pathway in high DFI sperm affects sperm capacitation","volume":"26","author":"Zhang","year":"2024","journal-title":"Asian J. Androl."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1007\/s12015-019-09929-9","article-title":"The Progresses of Spermatogonial Stem Cells Sorting Using Fluorescence-Activated Cell Sorting","volume":"16","author":"Cai","year":"2020","journal-title":"Stem Cell Rev. Rep."},{"key":"ref_61","first-page":"7.44.1","article-title":"Flow cytometry of murine spermatocytes","volume":"72","author":"Gaysinskaya","year":"2015","journal-title":"Curr. Protoc. Cytom."},{"key":"ref_62","first-page":"e53379","article-title":"Step-specific Sorting of Mouse Spermatids by Flow Cytometry","volume":"31","author":"Simard","year":"2015","journal-title":"J. Vis. Exp."}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/16\/2\/350\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,26]],"date-time":"2026-01-26T11:35:08Z","timestamp":1769427308000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/16\/2\/350"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1,22]]},"references-count":62,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2026,1]]}},"alternative-id":["ani16020350"],"URL":"https:\/\/doi.org\/10.3390\/ani16020350","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,22]]}}}