The Effects of Di-(2-ethylhexyl)-phthalate Exposure on Fertilization and Embryonic Development In Vitro and Testicular Genomic Mutation In Vivo

• Published in: PloS one Vol. 7; no. 11; p. e50465
• Main Authors: Huang, Xue-Feng, Li, Yan, Gu, Yi-Hua, Liu, Miao, Xu, Yan, Yuan, Yao, Sun, Fei, Zhang, Hui-Qin, Shi, Hui-Juan
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 30.11.2012; Public Library of Science (PLoS)
• Subjects: Analysis; Animals; Apoptosis; Biology; Birth rate; Blastocyst - drug effects; Blastocysts; Cell culture; Deoxyribonucleic acid; Diethylhexyl Phthalate - pharmacology; DNA; Drug dosages; Embryo Transfer; Embryo, Mammalian; Embryogenesis; Embryonic development; Embryonic Development - drug effects; Embryonic growth stage; Embryos; Epidemiology; Experiments; Exposure; Family planning; Female; Fertilization; Fertilization - drug effects; Fertilization in Vitro; Gene expression; Gene mutation; Genetic aspects; Genomes; Genomics; Hazards; Health care; Humans; In vitro fertilization; In vivo methods and tests; Infertility; Laboratories; Levels; Liver; Male; Medical research; Medicine; Metabolites; Mice; Mice, Transgenic; Motility; Mutagenesis; Mutation; Oocytes; Oocytes - drug effects; Outdoor air quality; Oxidative stress; Phthalates; Plasticizers - pharmacology; Population; Pregnancy; Reproduction - drug effects; Reproduction - physiology; Reproductive health; Reproductive system; Rodents; Sperm; Sperm Motility - drug effects; Spermatozoa; Spermatozoa - drug effects; Studies; Testes; Testis - cytology; Testis - drug effects; Toxicity; Transgenic mice; China
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0050465

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.