Evaluation of the effects of ethinylestradiol on sexual differentiation in the olvas-GFP/STII-YI medaka (transgenic Oryzias latipes) strain as estimated by proliferative activity of germ cells

Takeshi Hano, Yuji Oshima, Masato Kinoshita, Minoru Tanaka, Noriko Mishima, Yuko Wakamatsu, Kenjiro Ozato, Yohei Shimasaki, Tsuneo Honjo

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We evaluated the effects of 17(-ethinylestradiol (EE2) on sexual differentiation in transgenic olvas-GFP/STII-YI medaka (Oryzias latipes) in terms of the proliferative activity of germ cells. This strain contains the green fluorescent protein (GFP) gene fused to the regulatory region of the medaka vasa gene, and germ cell-specific expression of GFP can be visualized in living (transparent) individuals. From 0 days post-hatch (0dph) onwards, juveniles were exposed to graded concentrations of EE2 (25.2-1710ng/L) for 35 days. The gonads of live specimens were monitored by measuring their size and calculating their GFP-fluorescence area. GFP-fluorescent area in control females was about 10 times that in control males at 10 days posthatch (dph) whereas the gonadal size of 10dph males that had been exposed to 158ng/L of EE2 significantly increased up to twice the size of control males, indicating that abnormal sexual differentiation towards female might occur in these individuals. Histological examination and identification of the sex-linked marker SL1 indicated that male to female sex reversal occurred at EE2 exposure ≥45.1ng/L at 35dph. These results suggest that observation of proliferative activity of germ cells in the olvas-GFP/STII-YI strain could be applied to facilitated screening fish model to detect adverse effects on sexual differentiation as early as 10dph juveniles.

Original languageEnglish
Pages (from-to)177-184
Number of pages8
JournalAquatic Toxicology
Issue number3-4
Publication statusPublished - Aug 1 2011


All Science Journal Classification (ASJC) codes

  • Aquatic Science
  • Health, Toxicology and Mutagenesis

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