{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,15]],"date-time":"2026-07-15T12:53:11Z","timestamp":1784119991274,"version":"3.55.0"},"reference-count":57,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2023,9,29]],"date-time":"2023-09-29T00:00:00Z","timestamp":1695945600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Andrology"],"published-print":{"date-parts":[[2024,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Acetaminophen and ibuprofen are widely administered to babies due to their presumed safety as over\u2010the\u2010counter drugs. However, no reports exist on the effects of cyclooxygenase inhibitors on undifferentiated spermatogonia and spermatogonial stem cells. Infancy represents a critical period for spermatogonial stem cell formation and disrupting spermatogonial stem cells or their precursors may be associated with infertility and testicular cancer formation.<\/jats:p><\/jats:sec><jats:sec><jats:title>Objectives<\/jats:title><jats:p>The goal of this study was to examine the molecular and functional impact of cyclooxygenase inhibition and silencing on early steps of undifferentiated spermatogonia (u spg) and spermatogonial stem cell development, to assess the potential reproductive risk of pharmaceutical cyclooxygenase inhibitors.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>The effects of cyclooxygenase inhibition were assessed using the mouse C18\u20104 undifferentiated juvenile spermatogonial cell line model, previously shown to include cells with spermatogonial stem cell features, by measuring prostaglandins, cell proliferation, and differentiation, using cyclooxygenase 1\u2010 and cyclooxygenase 2\u2010selective inhibitors NS398, celecoxib, and FR122047, acetaminophen, and ibuprofen. Cyclooxygenase 1 gene silencing was achieved using a stable short\u2010hairpin RNA approach and clone selection, then assessing gene and protein expression in RNA sequencing, quantitative real\u2010time polymerase chain reaction, and immunofluorescence studies.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Cyclooxygenase 2 inhibitors NS398 and celecoxib, as well as acetaminophen, but not ibuprofen, dose\u2010dependently decreased retinoic acid\u2010induced expression of the spg differentiation gene <jats:italic>Stra8<\/jats:italic>, while NS398 decreased the spg differentiation marker Kit, suggesting that cyclooxygenase 2 is positively associated with spg differentiation. In contrast, short\u2010hairpin RNA\u2010based cyclooxygenase 1 silencing in C18\u20104 cells altered cellular morphology and upregulated <jats:italic>Stra8<\/jats:italic> and <jats:italic>Kit<\/jats:italic>, implying that cyclooxygenase 1 prevented spg differentiation. Furthermore, RNA sequencing analysis of cyclooxygenase 1 knockdown cells indicated the activation of several signaling pathways including the TGFb, Wnt, and Notch pathways, compared to control C18\u20104 cells. Notch pathway genes were upregulated by selective cyclooxygenase inhibitors, acetaminophen and ibuprofen.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>We report that cyclooxygenase 1 and 2 differentially regulate undifferentiated spermatogonia\/spermatogonial stem cell differentiation. Cyclooxygenases regulate Notch3 expression, with the Notch pathway targeted by PGD2. These data suggest an interaction between the eicosanoid and Notch signaling pathways that may be critical for the development of spermatogonial stem cells and subsequent spermatogenesis, cautioning about using cyclooxygenase inhibitors in infants.<\/jats:p><\/jats:sec>","DOI":"10.1111\/andr.13537","type":"journal-article","created":{"date-parts":[[2023,9,29]],"date-time":"2023-09-29T11:45:02Z","timestamp":1695987902000},"page":"899-917","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Differential roles of cyclooxygenase enzymes in the regulation of murine juvenile undifferentiated spermatogonia"],"prefix":"10.1111","volume":"12","author":[{"given":"Amy","family":"Tran\u2010Guzman","sequence":"first","affiliation":[{"name":"Department of Pharmacology and Pharmaceutical Sciences Alfred E. Mann School of Pharmacy and Pharmaceutical Sciences University of Southern California  Los Angeles California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amina","family":"Khan","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Pharmaceutical Sciences Alfred E. Mann School of Pharmacy and Pharmaceutical Sciences University of Southern California  Los Angeles California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8295-7716","authenticated-orcid":false,"given":"Martine","family":"Culty","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Pharmaceutical Sciences Alfred E. Mann School of Pharmacy and Pharmaceutical Sciences University of Southern California  Los Angeles California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2023,9,29]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrendo.2016.55"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajog.2011.11.019"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1289\/ehp2307"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365\u20102605.2012.01282.x"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0062556"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1034\/j.1600\u20100463.2003.11101031.x"},{"issue":"3","key":"e_1_2_10_8_1","doi-asserted-by":"crossref","first-page":"R139","DOI":"10.1530\/REP-14-0431","article-title":"Mammalian gonocyte and spermatogonia differentiation: recent advances and remaining challenges","volume":"149","author":"Manku G","year":"2015","journal-title":"Reproduction"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365\u20102605.2011.01199.x"},{"issue":"5","key":"e_1_2_10_10_1","doi-asserted-by":"crossref","first-page":"1063","DOI":"10.1213\/ANE.0b013e31828a4b54","article-title":"Nonsteroidal anti\u2010inflammatory drugs during pregnancy and the initiation of lactation","volume":"116","author":"Bloor M","year":"2013","journal-title":"Anesth Analg"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1210\/endo.142.7.8307"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1124\/pr.56.3.3"},{"issue":"3","key":"e_1_2_10_13_1","doi-asserted-by":"crossref","first-page":"691","DOI":"10.1111\/andr.12727","article-title":"Cyclooxygenase 2 (COX2) expression and prostaglandin synthesis in neonatal rat testicular germ cells: effects of acetaminophen and ibuprofen","volume":"8","author":"Manku G","year":"2020","journal-title":"Andrology"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.reprotox.2021.12.007"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ydbio.2004.12.006"},{"issue":"1","key":"e_1_2_10_16_1","doi-asserted-by":"crossref","first-page":"D377","DOI":"10.1093\/nar\/gks1118","article-title":"PANTHER in 2013: modeling the evolution of gene function, and other gene attributes, in the context of phylogenetic trees","volume":"41","author":"Mi H","year":"2012","journal-title":"Nucleic Acids Res"},{"issue":"9","key":"e_1_2_10_17_1","doi-asserted-by":"crossref","first-page":"2129","DOI":"10.1101\/gr.772403","article-title":"PANTHER: a library of protein families and subfamilies indexed by function","volume":"13","author":"Thomas PD","year":"2003","journal-title":"Genome Res"},{"issue":"1","key":"e_1_2_10_18_1","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/0090-6980(94)90074-4","article-title":"NS\u2010398, a new anti\u2010inflammatory agent, selectively inhibits prostaglandin G\/H synthase\/cyclooxygenase (COX\u20102) activity in vitro","volume":"47","author":"Futaki N","year":"1994","journal-title":"Prostaglandins"},{"issue":"9","key":"e_1_2_10_19_1","doi-asserted-by":"crossref","first-page":"1347","DOI":"10.1021\/jm960803q","article-title":"Synthesis and biological evaluation of the 1, 5\u2010diarylpyrazole class of cyclooxygenase\u20102 inhibitors: identification of 4\u2010[5\u2010(4\u2010methylphenyl)\u20103\u2010(trifluoromethyl)\u20101 H\u2010pyrazol\u20101\u2010yl] benzenesulfonamide (SC\u201058635, celecoxib)","volume":"40","author":"Penning TD","year":"1997","journal-title":"J Med Chem"},{"issue":"1","key":"e_1_2_10_20_1","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/S0014-2999(00)00051-0","article-title":"The analgesic effect profile of FR122047, a selective cyclooxygenase\u20101 inhibitor, in chemical nociceptive models","volume":"391","author":"Ochi T","year":"2000","journal-title":"Eur J Pharmacol"},{"issue":"12","key":"e_1_2_10_21_1","doi-asserted-by":"crossref","first-page":"1679","DOI":"10.1211\/0022357011778070","article-title":"Cyclooxygenase\u20101 and cyclooxygenase\u20102 selectivity of non\u2010steroidal anti\u2010inflammatory drugs: investigation using human peripheral monocytes","volume":"53","author":"Kato M","year":"2001","journal-title":"J Pharm Pharmacol"},{"issue":"2","key":"e_1_2_10_22_1","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1096\/fj.07-8506com","article-title":"Acetaminophen (paracetamol) is a selective cyclooxygenase\u20102 inhibitor in man","volume":"22","author":"Hinz B","year":"2008","journal-title":"FASEB J"},{"issue":"3","key":"e_1_2_10_23_1","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1002\/j.1552-4604.1992.tb03831.x","article-title":"Single\u2010dose pharmacokinetics of ibuprofen and acetaminophen in febrile children","volume":"32","author":"Brown RD","year":"1992","journal-title":"J Clin Pharmacol"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.envpol.2013.07.046"},{"issue":"1","key":"e_1_2_10_25_1","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1146\/annurev.biochem.69.1.145","article-title":"Cyclooxygenases: structural, cellular, and molecular biology","volume":"69","author":"Smith WL","year":"2000","journal-title":"Ann Rev Biochem"},{"issue":"1","key":"e_1_2_10_26_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1002\/bdrc.20142","article-title":"Gonocytes, the forgotten cells of the germ cell lineage","volume":"87","author":"Culty M","year":"2009","journal-title":"Birth Defects Res Part C: Embryo Today: Rev"},{"key":"e_1_2_10_27_1","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1146\/annurev.cellbio.24.110707.175355","article-title":"Regulation of spermatogonial stem cell self\u2010renewal in mammals","volume":"24","author":"Oatley JM","year":"2008","journal-title":"Ann Rev Cell Dev Biol"},{"issue":"1","key":"e_1_2_10_28_1","first-page":"1","article-title":"Developmental kinetics and transcriptome dynamics of stem cell specification in the spermatogenic lineage","volume":"10","author":"Law NC","year":"2019","journal-title":"Nat Commun"},{"issue":"6","key":"e_1_2_10_29_1","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1046\/j.1399-0004.2003.00194.x","article-title":"Notch signaling in development and disease","volume":"64","author":"Harper JA","year":"2003","journal-title":"Clin Genetics"},{"issue":"2","key":"e_1_2_10_30_1","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1002\/bies.20187","article-title":"Turning it up a Notch: cross\u2010talk between TGF\u03b2 and Notch signaling","volume":"27","author":"Kl\u00fcppel M","year":"2005","journal-title":"Bioessays"},{"issue":"1","key":"e_1_2_10_31_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3390\/jcm7010001","article-title":"Epithelial mesenchymal transition in embryonic development, tissue repair and cancer: a comprehensive overview","volume":"7","author":"Kim DH","year":"2018","journal-title":"J Clin Med"},{"issue":"8","key":"e_1_2_10_32_1","doi-asserted-by":"crossref","first-page":"900","DOI":"10.1634\/theoncologist.2017-0677","article-title":"The role of notch3 in cancer","volume":"23","author":"Aburjania Z","year":"2018","journal-title":"Oncologist"},{"key":"e_1_2_10_33_1","doi-asserted-by":"publisher","DOI":"10.1155\/2019\/8707053"},{"issue":"1","key":"e_1_2_10_34_1","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1080\/19336918.2019.1619958","article-title":"N3ICD with the transmembrane domain can effectively inhibit EMT by correcting the position of tight\/adherens junctions","volume":"13","author":"Tan J","year":"2019","journal-title":"Cell Adhesion Migration"},{"issue":"11","key":"e_1_2_10_35_1","doi-asserted-by":"crossref","first-page":"e269","DOI":"10.1038\/oncsis.2016.67","article-title":"Notch3 inhibits epithelial\u2013mesenchymal transition by activating Kibra\u2010mediated Hippo\/YAP signaling in breast cancer epithelial cells","volume":"5","author":"Zhang X","year":"2016","journal-title":"Oncogenesis"},{"issue":"5","key":"e_1_2_10_36_1","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1159\/000481772","article-title":"Expression profile of NOTCH3 in mouse Spermatogonia","volume":"204","author":"Okada R","year":"2017","journal-title":"Cells Tissues Organs"},{"issue":"7","key":"e_1_2_10_37_1","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0071213","article-title":"NOTCH1 gain of function in germ cells causes failure of spermatogenesis in male mice","volume":"8","author":"Huang Z","year":"2013","journal-title":"PLoS One"},{"issue":"1","key":"e_1_2_10_38_1","doi-asserted-by":"crossref","first-page":"188","DOI":"10.1016\/j.ydbio.2013.01.031","article-title":"Constitutive activation of NOTCH1 signaling in Sertoli cells causes gonocyte exit from quiescence","volume":"377","author":"Garcia TX","year":"2013","journal-title":"Dev Biol"},{"issue":"16","key":"e_1_2_10_39_1","doi-asserted-by":"crossref","first-page":"2538","DOI":"10.4161\/cc.25627","article-title":"NOTCH signaling in Sertoli cells regulates gonocyte fate","volume":"12","author":"Garcia TX","year":"2013","journal-title":"Cell cycle"},{"issue":"7","key":"e_1_2_10_40_1","doi-asserted-by":"crossref","first-page":"8423","DOI":"10.1096\/fj.201802361R","article-title":"Undifferentiated spermatogonia regulate Cyp26b1 expression through NOTCH signaling and drive germ cell differentiation","volume":"33","author":"Parekh PA","year":"2019","journal-title":"FASEB J"},{"key":"e_1_2_10_41_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.semcdb.2015.10.029"},{"issue":"4","key":"e_1_2_10_42_1","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1111\/j.1365-2605.2007.00785.x","article-title":"The transforming growth factor\u2010\u03b2 superfamily in early spermatogenesis: potential relevance to testicular dysgenesis","volume":"30","author":"Loveland KL","year":"2007","journal-title":"Int J Androl"},{"issue":"11","key":"e_1_2_10_43_1","doi-asserted-by":"crossref","first-page":"2227","DOI":"10.1093\/carcin\/bgn202","article-title":"COX\u20102 inactivates Smad signaling and enhances EMT stimulated by TGF\u2010\u03b2 through a PGE 2\u2010dependent mechanisms","volume":"29","author":"Neil JR","year":"2008","journal-title":"Carcinogenesis"},{"issue":"11","key":"e_1_2_10_44_1","doi-asserted-by":"crossref","first-page":"7238","DOI":"10.1074\/jbc.274.11.7238","article-title":"Delta\u2010induced Notch signaling mediated by RBP\u2010J inhibits MyoD expression and myogenesis","volume":"274","author":"Kuroda K","year":"1999","journal-title":"J Biol Chem"},{"issue":"21","key":"e_1_2_10_45_1","doi-asserted-by":"crossref","first-page":"8206","DOI":"10.1073\/pnas.83.21.8206","article-title":"Type beta transforming growth factor is an inhibitor of myogenic differentiation","volume":"83","author":"Massagu\u00e9 J","year":"1986","journal-title":"Proc Nat Acad Sci"},{"issue":"4","key":"e_1_2_10_46_1","doi-asserted-by":"crossref","first-page":"723","DOI":"10.1083\/jcb.200305112","article-title":"Cross\u2010talk between the Notch and TGF\u2010\u03b2 signaling pathways mediated by interaction of the Notch intracellular domain with Smad3","volume":"163","author":"Blokzijl A","year":"2003","journal-title":"J Cell Biol"},{"key":"e_1_2_10_47_1","doi-asserted-by":"publisher","DOI":"10.1242\/dev.00834"},{"issue":"3","key":"e_1_2_10_48_1","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1038\/sj.emboj.7600065","article-title":"Synergy and antagonism between Notch and BMP receptor signaling pathways in endothelial cells","volume":"23","author":"Itoh F","year":"2004","journal-title":"EMBO J"},{"issue":"12","key":"e_1_2_10_49_1","doi-asserted-by":"crossref","first-page":"5039","DOI":"10.1158\/0008-5472.CAN-08-4021","article-title":"The activated Notch1 signal pathway is associated with gastric cancer progression through cyclooxygenase\u20102","volume":"69","author":"Yeh T\u2010S","year":"2009","journal-title":"Cancer Res"},{"issue":"1","key":"e_1_2_10_50_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s42003-020-0904-6","article-title":"Prostaglandin EP2 receptor downstream of Notch signaling inhibits differentiation of human skeletal muscle progenitors in differentiation conditions","volume":"3","author":"Sakai\u2010Takemura F","year":"2020","journal-title":"Commun Biol"},{"key":"e_1_2_10_51_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep44184"},{"issue":"1","key":"e_1_2_10_52_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/srep19789","article-title":"Analgesic exposure in pregnant rats affects fetal germ cell development with inter\u2010generational reproductive consequences","volume":"6","author":"Dean A","year":"2016","journal-title":"Sci Rep"},{"issue":"18","key":"e_1_2_10_53_1","doi-asserted-by":"crossref","first-page":"3561","DOI":"10.1242\/dev.103408","article-title":"Prostaglandin D2 acts through the Dp2 receptor to influence male germ cell differentiation in the foetal mouse testis","volume":"141","author":"Moniot B","year":"2014","journal-title":"Development"},{"issue":"7","key":"e_1_2_10_54_1","doi-asserted-by":"crossref","first-page":"1890","DOI":"10.1093\/humrep\/det112","article-title":"aspirin and indomethacin display endocrine disrupting properties in the adult human testis in vitro","volume":"28","author":"Albert O","year":"2013","journal-title":"Human Reprod"},{"key":"e_1_2_10_55_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1582\u20104934.2010.01242.x"},{"key":"e_1_2_10_56_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mce.2008.02.018"},{"key":"e_1_2_10_57_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365\u20102605.2005.00643.x"},{"key":"e_1_2_10_58_1","doi-asserted-by":"publisher","DOI":"10.1053\/j.seminoncol.2009.07.006"}],"container-title":["Andrology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/andr.13537","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,4,11]],"date-time":"2024-04-11T10:12:19Z","timestamp":1712830339000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/andr.13537"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,9,29]]},"references-count":57,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2024,5]]}},"alternative-id":["10.1111\/andr.13537"],"URL":"https:\/\/doi.org\/10.1111\/andr.13537","archive":["Portico"],"relation":{},"ISSN":["2047-2919","2047-2927"],"issn-type":[{"value":"2047-2919","type":"print"},{"value":"2047-2927","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,9,29]]},"assertion":[{"value":"2023-03-15","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2023-09-10","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2023-09-29","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}