Effects of mono-(2-ethylhexyl) phthalate on fetal and neonatal rat testis organ cultures.
Di-(2-ethylhexyl) phthalate (DEHP) and its active metabolite, mono-(2-ethylhexyl) phthalate (MEHP), have been shown to cause reproductive toxicity in both developing and adult animals. In this study, we used organ cultures of fetal and neonatal rat testes to assess the in vitro effect of MEHP on seminiferous cord formation in Embryonic Day 13 (E13) testes and on the development of E18 and Postnatal Day 3 (P3) testes. Interestingly, MEHP had no effect on cord formation in the organ cultures of E13 testes, indicating that it has no effect on sexual differentiation of the indifferent gonad to testis. Consistently, the expression of a Sertoli cell-specific protein, mullerian inhibiting substance (MIS), or the number of gonocytes did not change in E13 testes after MEHP treatment. In contrast, MEHP decreased the levels of MIS and GATA-4 proteins in Sertoli cells and impaired Sertoli cell proliferation in the organ cultures of E18 and P3 testes. These results suggest that MEHP negatively influences proliferation and differentiation of Sertoli cells in both fetal and neonatal testes. In addition, MEHP treatment did not alter the number of gonocytes in E18 testes, whereas the number of gonocytes in P3 testes decreased in a dose-dependent manner, apparently due to enhanced apoptosis. These results suggest that MEHP adversely affects the gonocytes, which are mitotically active and undergoing migration and differentiation in neonatal testes, but it has no effect on fetal gonocytes that are mitotically quiescent.
Li
Hui
H
School of Molecular Biosciences, Center for Reproductive Biology, Washington State University, Pullman, Washington 99164, USA.
Kim
Kwan Hee
KH
D000818:Animals; D000831:Animals, Newborn; D054304:Anti-Mullerian Hormone; D017209:Apoptosis / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D053487:DEAD-box RNA Helicases; D004268:DNA-Binding Proteins / Q000187:drug effects / Q000378:metabolism; D004051:Diethylhexyl Phthalate / Q000031:analogs & derivatives* / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D005260:Female; D050991:GATA4 Transcription Factor; D006023:Glycoproteins / Q000187:drug effects / Q000378:metabolism; D008297:Male; D009924:Organ Culture Techniques / Q000379:methods; D020365:RNA Helicases / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012708:Sertoli Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D013735:Testicular Hormones / Q000378:metabolism; D013737:Testis / Q000166:cytology / Q000187:drug effects* / Q000196:embryology / Q000254:growth & development; D014157:Transcription Factors / Q000187:drug effects / Q000378:metabolism
D004268:DNA-Binding Proteins; D050991:GATA4 Transcription Factor; D006023:Glycoproteins; D013735:Testicular Hormones; D014157:Transcription Factors; D054304:Anti-Mullerian Hormone; D004051:Diethylhexyl Phthalate; C090864:Ddx4 protein, mouse; D053487:DEAD-box RNA Helicases; D020365:RNA Helicases; C016599:mono-(2-ethylhexyl)phthalate