Di-(2-ethylhexyl) phthalate and bisphenol A exposure impairs mouse primordial follicle assembly in vitro.
Bisphenol-A (BPA) and diethylhexyl phthalate (DEHP) are estrogenic compounds widely used in commercial plastic products. Previous studies have shown that exposure to such compounds have adverse effects on various aspects of mammalian reproduction including folliculogenesis. The objective of this study was to examine the effects of BPA and DEHP exposure on primordial follicle formation. We found that germ cell nest breakdown and primordial follicle assembly were significantly reduced when newborn mouse ovaries were exposed to 10 or 100 μM BPA and DEHP in vitro. Moreover, BPA and DEHP exposure increased the number of TUNEL positive oocytes and the mRNA level of the pro-apoptotic gene Bax in oocytes. These effects were associated with decreased expression of oocyte specific genes such as LIM homeobox 8 (Lhx8), factor in the germline alpha (Figla), spermatogenesis and oogenesis helix-loop-helix (Sohlh2), and newborn ovary homeobox (Nobox). Interestingly, BPA and DEHP exposure also prevented DNA demethylation of CpG sites of the Lhx8 gene in oocytes, a process normally associated with folliculogenesis. Finally, folliculogenesis was severely impaired in BPA and DEHP exposed ovaries after transplantation into the kidney capsules of immunodeficient mice. In conclusion, BPA and DEHP exposures impair mouse primordial follicle assembly in vitro.
Laboratory of Germ Cell Biology, Key Laboratory of Animal Reproduction and Germplasm Enhancement in Universities of Shandong, College of Animal Science and Technology, Qingdao Agricultural University, Qingdao, China.
Teng
T
Zhang
Lan
L
Li
Xun-Si
XS
Qin
Yang
Y
Zhou
Xi-Feng
XF
Zhang
Lin-Qing
LQ
Wang
Massimo
M
De Felici
Hong
H
Chen
Guo-Qing
GQ
Qin
Wei
W
Shen
D000818:Animals; D000831:Animals, Newborn; D001559:Benzhydryl Compounds / Q000633:toxicity*; D015153:Blotting, Western; D002478:Cells, Cultured; D004051:Diethylhexyl Phthalate / Q000633:toxicity*; D005260:Female; D016166:Free Radical Scavengers / Q000633:toxicity; D007124:Immunoenzyme Techniques; D066298:In Vitro Techniques; D051379:Mice; D016513:Mice, SCID; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D009866:Oogenesis / Q000187:drug effects*; D006080:Ovarian Follicle / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D010636:Phenols / Q000633:toxicity*; D010968:Plasticizers / Q000633:toxicity*; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000098826:Bisphenol A Compounds
D001559:Benzhydryl Compounds; D004051:Diethylhexyl Phthalate; D016166:Free Radical Scavengers; D010636:Phenols; D010968:Plasticizers; D012333:RNA, Messenger; C006780:bisphenol A; D000098826:Bisphenol A Compounds
DNA methylation; Lhx8; bisphenol-A; di-(2-ethylhexyl) phthalate; primordial follicle