{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T04:50:12Z","timestamp":1760244612362,"version":"build-2065373602"},"reference-count":28,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2022,11,25]],"date-time":"2022-11-25T00:00:00Z","timestamp":1669334400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ministry of Health &amp; Welfare, Republic of Korea","award":["HI21C1713","2019R1A2C1086882"],"award-info":[{"award-number":["HI21C1713","2019R1A2C1086882"]}]},{"DOI":"10.13039\/501100003725","name":"Basic Science Research Program through the National Research Foundation of Korea (NRF)","doi-asserted-by":"publisher","award":["HI21C1713","2019R1A2C1086882"],"award-info":[{"award-number":["HI21C1713","2019R1A2C1086882"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Non-obstructive azoospermia is a major clinical issue associated with male infertility that remains to be addressed. Although neogenin is reportedly abundantly expressed in the testis, its role in mammalian spermatogenesis is unknown. We systematically investigated the role of neogenin during spermatogenesis by performing loss-of-function studies. Testis-specific neogenin conditional knock-out (cKO) mice were generated using CRISPR\/Cas9 and neogenin-targeting guide RNAs. We analyzed the expression profiles of germ cell factors by RT-PCR and Western blotting. Neogenin localized mainly to spermatogonia in seminiferous tubules of mouse testes. RT-PCR and Western blot analyses further demonstrated that neogenin expression varied during spermatogenesis and was dramatically increased at postnatal day 12\u201325 during the pubertal stage. In neogenin-cKO mouse testes, the ratio of primary and secondary spermatocytes was significantly decreased compared with the control, while the number of apoptotic testicular cells was significantly increased. Taken together, these results suggest that neogenin plays a pivotal role in the maintenance and proliferation of spermatogonia during the early stage of spermatogenesis in mice.<\/jats:p>","DOI":"10.3390\/ijms232314761","type":"journal-article","created":{"date-parts":[[2022,11,28]],"date-time":"2022-11-28T01:58:46Z","timestamp":1669600726000},"page":"14761","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Down-Regulation of Neogenin Decreases Proliferation and Differentiation of Spermatogonia during the Early Phase of Spermatogenesis"],"prefix":"10.3390","volume":"23","author":[{"given":"Jin Woo","family":"Park","sequence":"first","affiliation":[{"name":"CHA Fertility Center Seoul Station, Seoul 04634, Republic of Korea"},{"name":"Department of Biomedical Sciences, College of Life Science, CHA University, Pochen 11160, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yu Jin","family":"Kim","sequence":"additional","affiliation":[{"name":"CHA Fertility Center Seoul Station, Seoul 04634, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sang Jin","family":"Lee","sequence":"additional","affiliation":[{"name":"Institute of Animal Genetic Resources Affiliated with Traditional Hanwoo Co., Ltd., Boryeong 33402, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jung Jae","family":"Ko","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences, College of Life Science, CHA University, Pochen 11160, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3237-6304","authenticated-orcid":false,"given":"Dae Keun","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Urology, CHA Fertility Center Seoul Station, CHA University School of Medicine, Seoul 04637, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8735-444X","authenticated-orcid":false,"given":"Jae Ho","family":"Lee","sequence":"additional","affiliation":[{"name":"CHA Fertility Center Seoul Station, Seoul 04634, Republic of Korea"},{"name":"Department of Biomedical Sciences, College of Life Science, CHA University, Pochen 11160, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,11,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1007\/s12522-016-0234-z","article-title":"Management of non-obstructive azoospermia","volume":"15","author":"Chiba","year":"2016","journal-title":"Reprod. Med. Biol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1007\/978-1-4614-4015-4_7","article-title":"Spermatogenesis","volume":"757","author":"Chu","year":"2013","journal-title":"Adv. Exp. Med. Biol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"e979623","DOI":"10.4161\/21565562.2014.979623","article-title":"Mechanisms of spermiogenesis and spermiation and how they are disturbed","volume":"4","year":"2014","journal-title":"Spermatogenesis"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1663","DOI":"10.1098\/rstb.2010.0026","article-title":"Spermatogonial stem cell regulation and spermatogenesis","volume":"365","author":"Phillips","year":"2010","journal-title":"Philos. Trans. R. Soc. Lond. B Biol. Sci."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"e28218","DOI":"10.4161\/spmg.28218","article-title":"Review of Azoospermia","volume":"4","author":"Wosnitzer","year":"2014","journal-title":"Spermatogenesis"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1838","DOI":"10.1016\/j.bbadis.2012.02.008","article-title":"Molecular control of rodent spermatogenesis","volume":"1822","author":"Jan","year":"2012","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1152\/physrev.00013.2015","article-title":"Spermatogenesis: The Commitment to Meiosis","volume":"96","author":"Griswold","year":"2016","journal-title":"Physiol. Rev."},{"key":"ref_8","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_9","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1146\/annurev.cellbio.24.110707.175355","article-title":"Regulation of spermatogonial stem cell self-renewal in mammals","volume":"24","author":"Oatley","year":"2008","journal-title":"Annu. Rev. Cell Dev. Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"874","DOI":"10.1016\/j.biocel.2006.10.023","article-title":"Neogenin: One receptor, many functions","volume":"39","author":"Wilson","year":"2007","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1369\/jhc.6A7031.2006","article-title":"Expression studies of neogenin and its ligand hemojuvelin in mouse tissues","volume":"55","author":"Rodriguez","year":"2007","journal-title":"J. Histochem. Cytochem."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1569","DOI":"10.1016\/j.biocel.2006.11.009","article-title":"Neogenin: A multi-functional receptor regulating diverse developmental processes","volume":"39","author":"Cole","year":"2007","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1016\/j.neuroscience.2006.06.041","article-title":"Characterization of neogenin-expressing neural progenitor populations and migrating neuroblasts in the embryonic mouse forebrain","volume":"142","author":"Fitzgerald","year":"2006","journal-title":"Neuroscience"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"458","DOI":"10.1038\/nsmb.3016","article-title":"Repulsive guidance molecule is a structural bridge between neogenin and bone morphogenetic protein","volume":"22","author":"Healey","year":"2015","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"5833","DOI":"10.1523\/JNEUROSCI.4487-15.2016","article-title":"Neogenin Promotes BMP2 Activation of YAP and Smad1 and Enhances Astrocytic Differentiation in Developing Mouse Neocortex","volume":"36","author":"Huang","year":"2016","journal-title":"J. Neurosci."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Lee, J.E., Kim, H.J., Bae, J.Y., Kim, S.W., Park, J.S., Shin, H.J., Han, W., Kim, S.W., Kang, K.S., and Noh, D.Y. (2005). Neogenin expression may be inversely correlated to the tumorigenicity of human breast cancer. BMC Cancer, 5.","DOI":"10.1186\/1471-2407-5-154"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"749","DOI":"10.1038\/ncb1157","article-title":"RGM and its receptor neogenin regulate neuronal survival","volume":"6","author":"Matsunaga","year":"2004","journal-title":"Nat. Cell Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"756","DOI":"10.1038\/ncb1156","article-title":"Neogenin mediates the action of repulsive guidance molecule","volume":"6","author":"Rajagopalan","year":"2004","journal-title":"Nat. Cell Biol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1038\/s41419-017-0019-2","article-title":"Neogenin, a regulator of adult hippocampal neurogenesis, prevents depressive-like behavior","volume":"9","author":"Sun","year":"2018","journal-title":"Cell Death Dis."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"500","DOI":"10.1007\/s11596-010-0457-x","article-title":"Effect of different concentrations of neogenin on proliferation, apoptosis and related proliferative factors in human trophoblasts","volume":"30","author":"Zhong","year":"2010","journal-title":"J. Huazhong Univ. Sci. Technol. Med. Sci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2928","DOI":"10.1634\/stemcells.2008-0134","article-title":"Spermatogonial stem cell self-renewal requires OCT4, a factor downregulated during retinoic acid-induced differentiation","volume":"26","author":"Dann","year":"2008","journal-title":"Stem Cells"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2516","DOI":"10.1634\/stemcells.2008-0379","article-title":"CD34+ testicular stromal cells support long-term expansion of embryonic and adult stem and progenitor cells","volume":"26","author":"Kim","year":"2008","journal-title":"Stem Cells"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Lee, J.H., Choi, S.S., Kim, H.W., Xiong, W.C., Min, C.K., and Lee, S.J. (2014). Neogenin as a receptor for early cell fate determination in preimplantation mouse embryos. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0101989"},{"key":"ref_24","first-page":"1","article-title":"Chromatin remodeling during mammalian spermatogenesis: Role of testis specific histone variants and transition proteins","volume":"63","author":"Pradeepa","year":"2007","journal-title":"Soc. Reprod. Fertil. Suppl."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1093\/molehr\/4.3.227","article-title":"The proliferation of spermatogonia in normal and pathological human seminiferous epithelium: An immunohistochemical study using monoclonal antibodies against Ki-67 protein and proliferating cell nuclear antigen","volume":"4","author":"Steger","year":"1998","journal-title":"Mol. Hum. Reprod."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"120","DOI":"10.1007\/BF00182039","article-title":"Paracrine factors and the regulation of spermatogenesis","volume":"11","author":"Niederberger","year":"1993","journal-title":"World J. Urol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"647","DOI":"10.1038\/ng1366","article-title":"Plzf is required in adult male germ cells for stem cell self-renewal","volume":"36","author":"Buaas","year":"2004","journal-title":"Nat. Genet."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"298","DOI":"10.1016\/j.contraception.2005.04.011","article-title":"Germ cell apoptosis control during spermatogenesis","volume":"72","author":"Giampietri","year":"2005","journal-title":"Contraception"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/23\/14761\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:27:00Z","timestamp":1760146020000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/23\/14761"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,11,25]]},"references-count":28,"journal-issue":{"issue":"23","published-online":{"date-parts":[[2022,12]]}},"alternative-id":["ijms232314761"],"URL":"https:\/\/doi.org\/10.3390\/ijms232314761","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2022,11,25]]}}}