{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,3]],"date-time":"2026-08-03T13:14:18Z","timestamp":1785762858729,"version":"3.56.0"},"reference-count":36,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2025,1,24]],"date-time":"2025-01-24T00:00:00Z","timestamp":1737676800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Key R&amp;D Program","award":["2022YFC2702800"],"award-info":[{"award-number":["2022YFC2702800"]}]},{"name":"National Key R&amp;D Program","award":["2021YFC2700302"],"award-info":[{"award-number":["2021YFC2700302"]}]},{"name":"National Key R&amp;D Program","award":["2021YFC2700501-1"],"award-info":[{"award-number":["2021YFC2700501-1"]}]},{"name":"National Key R&amp;D Program","award":["82221005"],"award-info":[{"award-number":["82221005"]}]},{"name":"National Key R&amp;D Program","award":["82122025"],"award-info":[{"award-number":["82122025"]}]},{"name":"National Key R&amp;D Program","award":["82201763"],"award-info":[{"award-number":["82201763"]}]},{"name":"National Key R&amp;D Program","award":["21KJA310007"],"award-info":[{"award-number":["21KJA310007"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["2022YFC2702800"],"award-info":[{"award-number":["2022YFC2702800"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["2021YFC2700302"],"award-info":[{"award-number":["2021YFC2700302"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["2021YFC2700501-1"],"award-info":[{"award-number":["2021YFC2700501-1"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82221005"],"award-info":[{"award-number":["82221005"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82122025"],"award-info":[{"award-number":["82122025"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82201763"],"award-info":[{"award-number":["82201763"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["21KJA310007"],"award-info":[{"award-number":["21KJA310007"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Natural Science Foundation of the Jiangsu Higher Education Institutions of China","award":["2022YFC2702800"],"award-info":[{"award-number":["2022YFC2702800"]}]},{"name":"Natural Science Foundation of the Jiangsu Higher Education Institutions of China","award":["2021YFC2700302"],"award-info":[{"award-number":["2021YFC2700302"]}]},{"name":"Natural Science Foundation of the Jiangsu Higher Education Institutions of China","award":["2021YFC2700501-1"],"award-info":[{"award-number":["2021YFC2700501-1"]}]},{"name":"Natural Science Foundation of the Jiangsu Higher Education Institutions of China","award":["82221005"],"award-info":[{"award-number":["82221005"]}]},{"name":"Natural Science Foundation of the Jiangsu Higher Education Institutions of China","award":["82122025"],"award-info":[{"award-number":["82122025"]}]},{"name":"Natural Science Foundation of the Jiangsu Higher Education Institutions of China","award":["82201763"],"award-info":[{"award-number":["82201763"]}]},{"name":"Natural Science Foundation of the Jiangsu Higher Education Institutions of China","award":["21KJA310007"],"award-info":[{"award-number":["21KJA310007"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Spermatogenesis is a complex process requiring precisely controlled metabolic adaptations. Although the genetic and cellular aspects of spermatogenesis have been extensively studied, the underlying metabolic mechanisms remain largely underexplored. In this study, we utilized STA-PUT technology to separate three key cell types involved in mouse spermatogenesis: pachytene spermatocytes (PAC), round spermatids (RS), and elongated spermatids (ES). A comprehensive untargeted metabolomic analysis revealed significant metabolic changes during spermatogenesis, such as reduced methylation-related metabolites and increased glycolytic intermediates and TCA cycle metabolites during ES. Moreover, metabolic differences between germ cells and somatic cells (Leydig and Sertoli cells) were highlighted, particularly in steroidogenesis and lipid metabolism. To investigate clinical relevance, we analyzed human seminal plasma. Samples from individuals with azoospermia displayed significant metabolic abnormalities, including reduced methionine, tryptophan, and arginine, which play vital roles in sperm development. Pathway enrichment analysis revealed disturbances in the metabolism of nucleotide, amino acid, and energy in azoospermia, suggesting potential biomarkers of male infertility. Our findings provide a comprehensive metabolic profile of spermatogenesis and suggest that metabolic alterations may be significant contributors to male infertility, particularly in cases of azoospermia.<\/jats:p>","DOI":"10.3390\/ijms26031001","type":"journal-article","created":{"date-parts":[[2025,1,24]],"date-time":"2025-01-24T08:48:23Z","timestamp":1737708503000},"page":"1001","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Characterization of Metabolic Patterns in Mouse Spermatogenesis and Its Clinical Implications in Humans"],"prefix":"10.3390","volume":"26","author":[{"given":"Jiachen","family":"Wang","sequence":"first","affiliation":[{"name":"State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing 211166, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mengqi","family":"Chen","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing 211166, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying","family":"Yao","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing 211166, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mengyuan","family":"Zhu","sequence":"additional","affiliation":[{"name":"Key Laboratory of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing 211166, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yingtong","family":"Jiang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing 211166, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiawei","family":"Duan","sequence":"additional","affiliation":[{"name":"Key Laboratory of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing 211166, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Yuan","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing 211166, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laihua","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing 211166, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5742-5080","authenticated-orcid":false,"given":"Minjian","family":"Chen","sequence":"additional","affiliation":[{"name":"Key Laboratory of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing 211166, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiahao","family":"Sha","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing 211166, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,1,24]]},"reference":[{"key":"ref_1","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_2","doi-asserted-by":"crossref","first-page":"2692","DOI":"10.1038\/nprot.2007.376","article-title":"Metabolic profiling, metabolomic and metabonomic procedures for NMR spectroscopy of urine, plasma, serum and tissue extracts","volume":"2","author":"Beckonert","year":"2007","journal-title":"Nat. Protoc."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1016\/j.jpba.2014.12.017","article-title":"Metabolomics for laboratory diagnostics","volume":"113","author":"Bujak","year":"2015","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"R55","DOI":"10.1530\/REP-21-0431","article-title":"Mitochondrial regulation in spermatogenesis","volume":"163","author":"Zhang","year":"2022","journal-title":"Reproduction"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1159\/000520662","article-title":"Metabolic control of germline formation and differentiation in mammals","volume":"16","author":"Hayashi","year":"2022","journal-title":"Sex. Dev."},{"key":"ref_6","first-page":"309","article-title":"Energy metabolism and sperm function","volume":"65","author":"Miki","year":"2007","journal-title":"Soc. Reprod. Fertil. Suppl."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"jcs235937","DOI":"10.1242\/jcs.235937","article-title":"Mitochondrial dynamics during spermatogenesis","volume":"133","author":"Varuzhanyan","year":"2020","journal-title":"J. Cell Sci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1586\/14737159.7.4.351","article-title":"Role of metabolomic analysis of biomarkers in the management of male infertility","volume":"7","author":"Deepinder","year":"2007","journal-title":"Expert Rev. Mol. Diagn."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"723","DOI":"10.1093\/humupd\/dmx023","article-title":"Metabolomics as a tool to identify biomarkers to predict and improve outcomes in reproductive medicine: A systematic review","volume":"23","author":"Saso","year":"2017","journal-title":"Hum. Reprod. Update"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1093\/molehr\/gaz056","article-title":"An optimised STAPUT method for the purification of mouse spermatocyte and spermatid populations","volume":"25","author":"Dunleavy","year":"2019","journal-title":"Mol. Hum. Reprod."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Jiang, H., Gao, Q., Zheng, W., Yin, S., Wang, L., Zhong, L., Ali, A., Khan, T., Hao, Q., and Fang, H. (2018). MOF influences meiotic expansion of H2AX phosphorylation and spermatogenesis in mice. PLoS Genet., 14.","DOI":"10.1371\/journal.pgen.1007300"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"103786","DOI":"10.1016\/j.isci.2022.103786","article-title":"Developmental alterations in DNA methylation during gametogenesis from primordial germ cells to sperm","volume":"25","author":"Maamar","year":"2022","journal-title":"IScience"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"R25","DOI":"10.1530\/REP-14-0643","article-title":"MicroRNAs and DNA methylation as epigenetic regulators of mitosis, meiosis and spermiogenesis","volume":"150","author":"Yao","year":"2015","journal-title":"Reproduction"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Yao, Y., Shi, B., Zhang, X., Wang, X., Li, S., Yao, Y., Guo, Y., Chen, D., Wang, B., and Yuan, Y. (2024). Germ cell-specific deletion of Pex3 reveals essential roles of PEX3-dependent peroxisomes in spermiogenesis. J. Biomed. Res., 38.","DOI":"10.7555\/JBR.37.20230055"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"954","DOI":"10.1095\/biolreprod.105.046896","article-title":"Mechanism of action of L-arginine on the vitality of spermatozoa is primarily through increased biosynthesis of nitric oxide","volume":"74","author":"Srivastava","year":"2006","journal-title":"Biol. Reprod."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1111\/j.1439-0272.2009.01005.x","article-title":"Activity of nitric oxide synthase in mature and immature human spermatozoa","volume":"42","author":"Roessner","year":"2010","journal-title":"Andrologia"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"564","DOI":"10.1210\/er.2014-1101","article-title":"The mammalian blood-testis barrier: Its biology and regulation","volume":"36","author":"Mruk","year":"2015","journal-title":"Endocr. Rev."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1002\/jemt.1070200104","article-title":"The blood-testis barrier and Sertoli cell junctions: Structural considerations","volume":"20","author":"Pelletier","year":"1992","journal-title":"Microsc. Res. Tech."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"903","DOI":"10.1111\/andr.12703","article-title":"Essential roles of interstitial cells in testicular development and function","volume":"8","author":"Heinrich","year":"2020","journal-title":"Andrology"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1016\/j.mce.2008.11.005","article-title":"Role of androgen and gonadotrophins in the development and function of the Sertoli cells and Leydig cells: Data from mutant and genetically modified mice","volume":"306","author":"Morris","year":"2009","journal-title":"Mol. Cell. Endocrinol."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Duan, N., Ran, Y., Wang, H., Luo, Y., Gao, Z., Lu, X., Cui, F., Chen, Q., Xue, B., and Liu, X. (2024). Mouse testicular macrophages can independently produce testosterone and are regulated by Cebpb. Biol. Res., 57.","DOI":"10.1186\/s40659-024-00544-8"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Gao, Y., Wang, Z., Long, Y., Yang, L., Jiang, Y., Ding, D., Teng, B., Chen, M., Yuan, J., and Gao, F. (2024). Unveiling the roles of Sertoli cells lineage differentiation in reproductive development and disorders: A review. Front. Endocrinol., 15.","DOI":"10.3389\/fendo.2024.1357594"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"152046","DOI":"10.1016\/j.acthis.2023.152046","article-title":"Immunohistochemical analysis of the vimentin filaments in Sertoli cells is a powerful tool for the prediction of spermatogenic dysfunction","volume":"125","author":"Saito","year":"2023","journal-title":"Acta Histochem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"4829180","DOI":"10.1155\/2017\/4829180","article-title":"Ethanolamine and phosphatidylethanolamine: Partners in health and disease","volume":"2017","author":"Patel","year":"2017","journal-title":"Oxidative Med. Cell. Longev."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"457","DOI":"10.1080\/15548627.2020.1720431","article-title":"m6A mRNA methylation regulates testosterone synthesis through modulating autophagy in Leydig cells","volume":"17","author":"Chen","year":"2021","journal-title":"Autophagy"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Krause, N., and Wegner, A. (2024). N-acetyl-aspartate metabolism at the interface of cancer, immunity, and neurodegeneration. Curr. Opin. Biotechnol., 85.","DOI":"10.1016\/j.copbio.2023.103051"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"152996","DOI":"10.1016\/j.tox.2021.152996","article-title":"3-Indolepropionic acid upturned male reproductive function by reducing oxido-inflammatory responses and apoptosis along the hypothalamic-pituitary-gonadal axis of adult rats exposed to chlorpyrifos","volume":"463","author":"Owumi","year":"2021","journal-title":"Toxicology"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Guerriero, G., Trocchia, S., Abdel-Gawad, F.K., and Ciarcia, G. (2014). Roles of reactive oxygen species in the spermatogenesis regulation. Front. Endocrinol., 5.","DOI":"10.3389\/fendo.2014.00056"},{"key":"ref_29","first-page":"109","article-title":"Metabolic fingerprinting of seminal plasma from non-obstructive azoospermia patients: Positive versus negative sperm retrieval","volume":"19","author":"Gilany","year":"2018","journal-title":"J. Reprod. Infertil."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1186\/s12610-023-00184-0","article-title":"Seminal plasma biomarkers for predicting successful sperm retrieval in patients with nonobstructive azoospermia: A narrative review of human studies","volume":"33","author":"Li","year":"2023","journal-title":"Basic Clin. Androl."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1859","DOI":"10.1111\/andr.12882","article-title":"Metabolomic signature of the seminal plasma in men with severe oligoasthenospermia","volume":"8","author":"Boguenet","year":"2020","journal-title":"Andrology"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Liang, J., Tang, Y., Li, H., Mei, J., Cao, Z., Xia, H., Huang, R., Yang, Y., and Huang, Y. (2021). Isolation of primary leydig cells from murine testis. Bio Protoc., 11.","DOI":"10.21769\/BioProtoc.4223"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/978-1-4939-7698-0_1","article-title":"Establishment of primary culture of sertoli cells","volume":"1748","author":"Bernardino","year":"2018","journal-title":"Sertoli Cells"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1093\/humupd\/dmp048","article-title":"World Health Organization reference values for human semen characteristics","volume":"16","author":"Cooper","year":"2010","journal-title":"Hum. Reprod. Update"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2847","DOI":"10.1093\/bioinformatics\/btw313","article-title":"Complex heatmaps reveal patterns and correlations in multidimensional genomic data","volume":"32","author":"Gu","year":"2016","journal-title":"Bioinformatics"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"4313","DOI":"10.1093\/bioinformatics\/bty528","article-title":"MetaboAnalystR: An R package for flexible and reproducible analysis of metabolomics data","volume":"34","author":"Chong","year":"2018","journal-title":"Bioinformatics"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/3\/1001\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,8]],"date-time":"2025-10-08T10:35:35Z","timestamp":1759919735000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/3\/1001"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,1,24]]},"references-count":36,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2025,2]]}},"alternative-id":["ijms26031001"],"URL":"https:\/\/doi.org\/10.3390\/ijms26031001","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,1,24]]}}}