Transcriptional alterations induced by binary mixtures of ethinylestradiol and norgestrel during the early development of zebrafish (Danio rerio)

Comp Biochem Physiol C Toxicol Pharmacol. 2017 May;195:60-67. doi: 10.1016/j.cbpc.2017.02.005. Epub 2017 Feb 17.


Synthetic estrogens and progestins are commonly used in human and veterinary medicine. After use, they reach aquatic environments via discharge of wastewaters from human and animals, thus posing potential risks to organisms. So far, very little is known about their combined effects in aquatic organisms. The aim of this study was to investigate the effects of binary mixtures of ethinylestradiol (EE2) and norgestrel (NGT) on embryonic zebrafish (Danio rerio) by measuring transcriptional alterations. Zebrafish embryos were exposed to EE2 and NGT alone or in combination at concentrations between 36 and 5513ngL-1 for 96h post-fertilization (hpf). The results showed that most of gene transcriptions of hypothalamic-pituitary-gonadal axis (e.g., Pgr, Mprα, Esr1, Esr2a, Vtg1, Ar, Cyp11b, Star, Gnrh3 and Fshb) and circadian rhythm signaling (e.g., Cry1a, Cry2a, Cry2b, Per3, Arntl1b, Arntl2, Clock1a, Cry3 and Cry4) displayed most pronounced alterations in the mixtures as compared to single EE2 and NGT exposures. This finding suggests exposure to the binary mixtures of EE2 and NGT produced significantly enhanced effects in fish as compared to single chemical exposures, and their coexistence could have significant environmental implications.

Keywords: Ethinylestradiol; Gene transcription; Mixture effects; Norgestrel.

MeSH terms

  • Analysis of Variance
  • Animals
  • Dose-Response Relationship, Drug
  • Drug Synergism
  • Estrogens / pharmacology
  • Ethinyl Estradiol / pharmacology*
  • Female
  • Gene Expression Profiling / methods*
  • Gene Expression Regulation, Developmental / drug effects*
  • Male
  • Norgestrel / pharmacology*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Time Factors
  • Zebrafish / embryology
  • Zebrafish / genetics*
  • Zebrafish Proteins / genetics


  • Estrogens
  • Zebrafish Proteins
  • Norgestrel
  • Ethinyl Estradiol