{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,29]],"date-time":"2026-06-29T17:47:29Z","timestamp":1782755249699,"version":"3.54.5"},"reference-count":20,"publisher":"Public Library of Science (PLoS)","issue":"6","license":[{"start":{"date-parts":[[2026,6,29]],"date-time":"2026-06-29T00:00:00Z","timestamp":1782691200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Corestemchemon"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>The spermatogenic stage serves as a vital criterion for assessing normal spermatogenesis and is central to evaluating reproductive toxicity. Current manual methods for evaluating the spermatogenic stage are time-intensive, require expert knowledge, and are less effective at detecting subtle changes or comparing stage frequencies across samples. To overcome these limitations, this study introduces a method that leverages object detection models and Region-based Convolutional Neural Networks to enable efficient and accurate evaluation of spermatogenic stages. A total of 16 periodic acid-Schiff-stained testicular tissue whole-slide images (WSIs) obtained from 16 Sprague\u2013Dawley rats were used in this study. A total of 14 stages were identified, and the approach was further applied to atrophied testicular samples as a real-world example. A total of 10 WSIs (nine normal and one atrophied testes) were used for model training, validation, and testing. Six additional WSIs (three normal and three atrophied testes) were used for model inference. For the test set, the model achieved a mean average precision of 0.869 and a mean average recall of 0.977 for detecting spermatogenic stages and atrophy. For the inference set, agreement with pathologist assessments exceeded 91%, providing objective benchmarks for stage evaluation and facilitating the comparison of stage frequencies across multiple samples. The model enabled the quantitative assessment of atrophied tissues by analyzing the proportional changes in atrophied seminiferous tubules relative to normal tubules. This automated approach has the potential to reduce the workload of pathologists by enabling rapid, reproducible assessment of toxicological changes during spermatogenesis. As a proof-of-concept, the integration of deep learning demonstrated the feasibility of improving the efficiency and objectivity of pathological evaluations in reproductive toxicity studies.<\/jats:p>","DOI":"10.1371\/journal.pone.0337245","type":"journal-article","created":{"date-parts":[[2026,6,29]],"date-time":"2026-06-29T17:27:16Z","timestamp":1782754036000},"page":"e0337245","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Automated spermatogenic staging in periodic acid-Schiff-stained testes of Sprague\u2013Dawley rats using a deep learning model for normal and atrophied tissues"],"prefix":"10.1371","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-0572-489X","authenticated-orcid":true,"given":"Da-Mi","family":"Kim","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jin-Hyung","family":"Rho","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"So-Young","family":"Wee","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4388-0952","authenticated-orcid":true,"given":"Hwa-Young","family":"Son","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,6,29]]},"reference":[{"issue":"1","key":"pone.0337245.ref001","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1080\/019262301301418856","article-title":"Overview of male reproductive pathology","volume":"29","author":"GL Foley","year":"2001","journal-title":"Toxicol Pathol"},{"issue":"4","key":"pone.0337245.ref002","doi-asserted-by":"crossref","first-page":"548","DOI":"10.1111\/j.1749-6632.1952.tb26576.x","article-title":"Definition of the stages of the cycle of the seminiferous epithelium in the rat","volume":"55","author":"CP Leblond","year":"1952","journal-title":"Ann N Y Acad Sci"},{"issue":"4","key":"pone.0337245.ref003","first-page":"451","article-title":"The stages of the cycle of the seminiferous epithelium of the rat: practical definitions in PA-Schiff-hematoxylin and hematoxylin-eosin stained sections","volume":"16","author":"Y Clermont","year":"1957","journal-title":"Rev Can Biol"},{"key":"pone.0337245.ref004","doi-asserted-by":"crossref","DOI":"10.1177\/0192623312454337","article-title":"Proliferative and nonproliferative lesions of the rat and mouse male reproductive system","volume":"40","author":"D Creasy","year":"2012","journal-title":"Toxicol Pathol"},{"issue":"3","key":"pone.0337245.ref005","doi-asserted-by":"crossref","first-page":"338","DOI":"10.1177\/019262339502300310","article-title":"Incidence of nonneoplastic lesions in historical control male and female Fischer-344 rats from 90-day toxicity studies","volume":"23","author":"D Dixon","year":"1995","journal-title":"Toxicol Pathol"},{"key":"pone.0337245.ref006","volume-title":"Test No. 421: Reproduction\/Developmental Toxicity Screening Test","author":"OECD","year":"2016"},{"key":"pone.0337245.ref007","volume-title":"Test No. 422: Combined Repeated Dose Toxicity Study with the Reproduction\/Developmental Toxicity Screening Test","author":"OECD","year":"2016"},{"key":"pone.0337245.ref008","doi-asserted-by":"crossref","first-page":"101835","DOI":"10.1016\/j.media.2020.101835","article-title":"Computerized spermatogenesis staging (CSS) of mouse testis sections via quantitative histomorphological analysis","volume":"70","author":"J Xu","year":"2021","journal-title":"Med Image Anal"},{"issue":"4","key":"pone.0337245.ref009","doi-asserted-by":"crossref","first-page":"872","DOI":"10.1177\/0192623320969678","article-title":"Deep learning-based spermatogenic staging assessment for hematoxylin and eosin-stained sections of rat testes","volume":"49","author":"DM Creasy","year":"2021","journal-title":"Toxicol Pathol"},{"issue":"6","key":"pone.0337245.ref010","doi-asserted-by":"crossref","first-page":"736","DOI":"10.1177\/01926233221117747","article-title":"Morphologic features and deep learning-based analysis of canine spermatogenic stages","volume":"50","author":"S Mehrvar","year":"2022","journal-title":"Toxicol Pathol"},{"issue":"1","key":"pone.0337245.ref011","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1177\/01926233241234059","article-title":"Deep learning\u2013based spermatogenic staging in tissue sections of cynomolgus macaque testes","volume":"52","author":"L Mecklenburg","year":"2024","journal-title":"Toxicol Pathol"},{"issue":"6","key":"pone.0337245.ref012","doi-asserted-by":"crossref","first-page":"1137","DOI":"10.1109\/TPAMI.2016.2577031","article-title":"Faster R-CNN: towards real-time object detection with region proposal networks","volume":"39","author":"S Ren","year":"2017","journal-title":"IEEE Trans Pattern Anal Mach Intell"},{"issue":"5","key":"pone.0337245.ref013","first-page":"1483","article-title":"IEEE transactions on pattern analysis and machine intelligence","volume":"43","author":"Z Cai","year":"2019","journal-title":"IEEE Trans Pattern Anal Mach Intel"},{"key":"pone.0337245.ref014","doi-asserted-by":"crossref","first-page":"27","DOI":"10.4103\/2153-3539.119005","article-title":"OpenSlide: a vendor-neutral software foundation for digital pathology","volume":"4","author":"A Goode","year":"2013","journal-title":"J Pathol Inform"},{"key":"pone.0337245.ref015","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1111\/j.1365-2605.1993.tb01156.x","article-title":"Histological and histopathological evaluation of the testis","volume":"16","author":"R Russell","year":"1993","journal-title":"Int J Androl"},{"key":"pone.0337245.ref016","article-title":"MMDetection: Open mmlab detection toolbox and benchmark. arXiv preprint arXiv: 190607155","author":"K Chen","year":"2019"},{"key":"pone.0337245.ref017","article-title":"SAHI: A lightweight vision library for performing large scale object detection and instance segmentation","author":"FC Akyon","year":"2021","journal-title":"Zenodo"},{"issue":"3","key":"pone.0337245.ref018","doi-asserted-by":"crossref","first-page":"517","DOI":"10.1095\/biolreprod43.3.517","article-title":"Frequency of the stages in the cycle of the seminiferous epithelium in the rat","volume":"43","author":"RA Hess","year":"1990","journal-title":"Biol Reprod"},{"issue":"6","key":"pone.0337245.ref019","doi-asserted-by":"crossref","first-page":"2608","DOI":"10.1210\/endo.135.6.7988449","article-title":"Testosterone promotes the conversion of round spermatids between stages VII and VIII of the rat spermatogenic cycle","volume":"135","author":"L O\u2019Donnell","year":"1994","journal-title":"Endocrinology"},{"issue":"4","key":"pone.0337245.ref020","doi-asserted-by":"crossref","first-page":"547","DOI":"10.1002\/ar.1092350407","article-title":"Stage-dependent changes in spermatogenesis and Sertoli cells in relation to the onset of spermatogenic failure following withdrawal of testosterone","volume":"235","author":"JB Kerr","year":"1993","journal-title":"Anat Rec"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0337245","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,29]],"date-time":"2026-06-29T17:27:22Z","timestamp":1782754042000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0337245"}},"subtitle":[],"editor":[{"given":"Suresh","family":"Yenugu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,6,29]]},"references-count":20,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2026,6,29]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0337245","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,29]]}}}