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Endocrinol."],"abstract":"<jats:p>Correct fetal testis development underpins adult male fertility, and TGF\u03b2 superfamily ligands control key aspects of this process. Transcripts encoding one such ligand, activin A, are upregulated in testes after sex determination and remain high until after birth. Testis development requires activin signalling; mice lacking activin A (<jats:italic>Inhba<\/jats:italic> KO) display altered somatic and germ cell proliferation, disrupted cord elongation and altered steroid synthesis. In human pregnancies with pre-eclampsia, the foetus is inappropriately exposed to elevated activin A. To learn how this affects testis development, we examined mice lacking the potent activin inhibitor, inhibin, (<jats:italic>Inha<\/jats:italic> KO) at E13.5, E15.5 and PND0. At E13.5, testes appeared similar in WT and KO littermates, however E15.5 <jats:italic>Inha<\/jats:italic> KO testes displayed two germline phenotypes: (1) multinucleated germ cells within cords, and (2) germ cells outside of cords, both of which are documented following <jats:italic>in utero<\/jats:italic> exposure to endocrine disrupting phthalates in rodents. Quantitation of Sertoli and germ cells in <jats:italic>Inha<\/jats:italic> KO (modelling elevated activin A) and <jats:italic>Inhba<\/jats:italic> KO (low activin A) testes using immunofluorescence demonstrated activin A bioactivity determines the Sertoli\/germ cell ratio. The 50% reduction in gonocytes in <jats:italic>Inha<\/jats:italic> KO testes at birth indicates unopposed activin A has a profound impact on embryonic germ cells. Whole testis RNAseq on <jats:italic>Inha<\/jats:italic> KO mice revealed most transcripts affected at E13.5 were present in Leydig cells and associated with steroid biosynthesis\/metabolism. In agreement, androstenedione (A4), testosterone (T), and the A4:T ratio were reduced in <jats:italic>Inha<\/jats:italic> KO testes at E17.5, confirming unopposed activin A disrupts testicular steroid production. E15.5 testes cultured with either activin A and\/or mono-2-ethylhexyl phthalate (MEHP) generated common histological and transcriptional outcomes affecting germline and Leydig cells, recapitulating the phenotype observed in <jats:italic>Inha<\/jats:italic> KO testes. Cultures with activin A and MEHP together provided evidence of common targets. Lastly, this study extends previous work focussed on the <jats:italic>Inhba<\/jats:italic> KO model to produce a signature of activin A bioactivity in the fetal testis. These outcomes show the potential for elevated activin A signalling to replicate some aspects of fetal phthalate exposure prior to the masculinization programming window, influencing fetal testis growth and increasing the risk of testicular dysgenesis.<\/jats:p>","DOI":"10.3389\/fendo.2023.1234712","type":"journal-article","created":{"date-parts":[[2023,9,1]],"date-time":"2023-09-01T08:24:31Z","timestamp":1693556671000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":7,"title":["Testis exposure to unopposed\/elevated activin A in utero affects somatic and germ cells and alters steroid levels mimicking phthalate exposure"],"prefix":"10.3389","volume":"14","author":[{"given":"Penny A. F.","family":"Whiley","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael C. 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