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In this study, we systematically described testicular development and identified thirteen different germ cell types throughout the reproductive cycle in large yellow croaker (Larimichthys crocea), a commercially important marine cultured fish in East Asia. Using a set of specific antibodies (VASA, PCNA, DMC1, NANOS2 and GSDF), we developed a high-throughput immunohistochemistry method to identify different types of spermatogenic cells, with a particular focus on distinguishing spermatogonial subtypes. VASA was strongly expressed in all four types of spermatogonia (As, Apr, Adiff, and B) and decreased progressively during spermatogenesis. DMC1 exhibited distinct expression patterns in different spermatocytes subtypes, and GSDF was highly expressed in somatic cells surrounding type A spermatogonia. Particularly, NANOS2 was highly specific to As and Apr spermatogonia, supporting their role as SSC candidates. By morphological observation and co-staining of VASA and PCNA, we found that As spermatogonia exhibited dynamic development characteristics during the annual reproductive cycle. These findings provide a valuable tool for reproductive studies and potential applications in surrogate reproduction through SSCs transplantation in teleost fish.<\/jats:p>","DOI":"10.1093\/biolre\/ioaf065","type":"journal-article","created":{"date-parts":[[2025,4,10]],"date-time":"2025-04-10T12:17:55Z","timestamp":1744287475000},"page":"1185-1199","source":"Crossref","is-referenced-by-count":6,"title":["Morphological characterization of spermatogenesis and spermatogonial stem cells in <i>Larimichthys crocea<\/i>, a seasonal breeding teleost"],"prefix":"10.1093","volume":"112","author":[{"given":"Yinan","family":"Zhou","sequence":"first","affiliation":[{"name":"School of Fisheries, Zhejiang Ocean University , Zhoushan, Zhejiang ,","place":["China"]},{"name":"Marine and Fishery Research Institute , Zhoushan, Zhejiang ,","place":["China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yang","family":"Yang","sequence":"additional","affiliation":[{"name":"Marine and Fishery Research Institute , Zhoushan, Zhejiang ,","place":["China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Huan","family":"Ye","sequence":"additional","affiliation":[{"name":"Key Laboratory of Freshwater Biodiversity Conservation, Ministry of Agriculture of China, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences , Wuhan, Hubei ,","place":["China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lulu","family":"Mi","sequence":"additional","affiliation":[{"name":"School of Life Sciences, Institute of Life Sciences and Green Development, Hebei University , Baoding, Hebei ,","place":["China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Weihua","family":"Hu","sequence":"additional","affiliation":[{"name":"Marine and Fishery Research Institute , Zhoushan, Zhejiang ,","place":["China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dongdong","family":"Xu","sequence":"additional","affiliation":[{"name":"Marine and Fishery Research Institute , Zhoushan, Zhejiang ,","place":["China"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2025,4,10]]},"reference":[{"key":"2025062516103700800_ref1","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1016\/j.ygcen.2009.02.013","article-title":"Spermatogenesis in fish","volume":"165","author":"Schulz","year":"2010","journal-title":"Gen Comp Endocr"},{"key":"2025062516103700800_ref2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1095\/biolreprod.113.112391","article-title":"Spermatogenic cycle length and sperm production in the freshwater turtle Kinosternon scorpioides","volume":"90","author":"Sousa","year":"2014","journal-title":"Biol 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