title: The effects of Di-(2-ethylhexyl)-phthalate exposure on fertilization and embryonic development in vitro and testicular genomic mutation in vivo.
abstract: The present study was undertaken to determine the reproductive hazards of Di-(2-ethylhexyl)-phthalate (DEHP) on mouse spermatozoa and embryos in vitro and genomic changes in vivo. Direct low-level DEHP exposure (1 μg/ml) on spermatozoa and embryos was investigated by in vitro fertilization (IVF) process, culture of preimplanted embryos in DEHP-supplemented medium and embryo transfer to achieve full term development. Big Blue® transgenic mouse model was employed to evaluate the mutagenesis of testicular genome with in vivo exposure concentration of DEHP (500 mg/kg/day). Generally, DEHP-treated spermatozoa (1 μg/ml, 30 min) presented reduced fertilization ability (P<0.05) and the resultant embryos had decreased developmental potential compared to DMSO controls (P<0.05). Meanwhile, the transferred 2-cell stage embryos derived from treated spermatozoa also exhibited decreased birth rate than that of control (P<0.05). When fertilized oocytes or 2-cell stage embryos were recovered by in vivo fertilization (without treatment) and then exposed to DEHP, the subsequent development proceed to blastocysts was different, fertilized oocytes were significantly affected (P<0.05) whereas developmental progression of 2-cell stage embryos was similar to controls (P>0.05). Testes of the Big Blue® transgenic mice treated with DEHP for 4 weeks indicated an approximately 3-fold increase in genomic DNA mutation frequency compared with controls (P<0.05). These findings unveiled the hazardous effects of direct low-level exposure of DEHP on spermatozoa's fertilization ability as well as embryonic development, and proved that in vivo DEHP exposure posed mutagenic risks in the reproductive organ - at least in testes, are of great concern to human male reproductive health.
authors.affiliation: Shanghai Medical College, Fudan University, Shanghai, China. xuefhuang@hotmail.com
authors.forename: Xue-Feng
authors.initials: XF
authors.lastname: Huang
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Yi-Hua
authors.initials: YH
authors.lastname: Gu
authors.forename: Miao
authors.initials: M
authors.lastname: Liu
authors.forename: Yan
authors.initials: Y
authors.lastname: Xu
authors.forename: Yao
authors.initials: Y
authors.lastname: Yuan
authors.forename: Fei
authors.initials: F
authors.lastname: Sun
authors.forename: Hui-Qin
authors.initials: HQ
authors.lastname: Zhang
authors.forename: Hui-Juan
authors.initials: HJ
authors.lastname: Shi
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000187:drug effects*; D004051:Diethylhexyl Phthalate / Q000494:pharmacology*; D004624:Embryo Transfer; D004622:Embryo, Mammalian; D047108:Embryonic Development / Q000187:drug effects*; D005260:Female; D005306:Fertilization / Q000187:drug effects*; D005307:Fertilization in Vitro; D006801:Humans; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009154:Mutation; D009865:Oocytes / Q000187:drug effects*; D010968:Plasticizers / Q000494:pharmacology*; D011247:Pregnancy; D012098:Reproduction / Q000187:drug effects / Q000502:physiology; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects*; D013737:Testis / Q000166:cytology / Q000187:drug effects*
chemical_list: D010968:Plasticizers; D004051:Diethylhexyl Phthalate