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Advanced fluorescence in situ hybridization to localize and quantify gene expression in Japanese medaka (Oryzias latipes) exposed to endocrine-disrupting compounds

Authors

  • June-Woo Park,

    Corresponding author
    1. Department of Zoology, National Food Safety and Toxicology Center, Michigan State University, East Lansing, Michigan 48824, USA
    Current affiliation:
    1. Park is Center for Environmental Biotechnology, University of Tennessee, Knoxville, TN 37996, USA
    • Department of Zoology, National Food Safety and Toxicology Center, Michigan State University, East Lansing, Michigan 48824, USA
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  • Amber R. Tompsett,

    1. Department of Biomedical Veterinary Sciences and Toxicology Centre, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5B3, Canada
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  • Xiaowei Zhang,

    1. Department of Biomedical Veterinary Sciences and Toxicology Centre, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5B3, Canada
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  • John L. Newsted,

    1. Department of Zoology, National Food Safety and Toxicology Center, Michigan State University, East Lansing, Michigan 48824, USA
    2. ENTRIX, Okemos, Michigan 48823, USA
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  • Paul D. Jones,

    1. Department of Biomedical Veterinary Sciences and Toxicology Centre, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5B3, Canada
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  • Doris W. T. Au,

    1. Department of Biomedical Veterinary Sciences and Toxicology Centre, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5B3, Canada
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  • Richard Kong,

    1. Department of Biology and Chemistry, City University of Hong Kong, Kowloon, Hong Kong, Special Administrative Region of China, China
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  • Rudolf S. S. Wu,

    1. Department of Biology and Chemistry, City University of Hong Kong, Kowloon, Hong Kong, Special Administrative Region of China, China
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  • John P. Giesy,

    1. Department of Zoology, National Food Safety and Toxicology Center, Michigan State University, East Lansing, Michigan 48824, USA
    2. Department of Biology and Chemistry, City University of Hong Kong, Kowloon, Hong Kong, Special Administrative Region of China, China
    3. School of Environmental Science, Nanjing University, Nanjing, China
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  • Markus Hecker

    1. Department of Zoology, National Food Safety and Toxicology Center, Michigan State University, East Lansing, Michigan 48824, USA
    2. Department of Biomedical Veterinary Sciences and Toxicology Centre, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5B3, Canada
    3. ENTRIX, Saskatoon, Saskatchewan S7N 5B3, Canada
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Abstract

In an earlier study, we described the development of fluorescence in situ hybridization (FISH) using confocal microscopy to localize and quantify gene expression in fish. Here, we report the results of FISH application to investigate effects of model endocrine-disrupting chemicals (EDCs), 17α-ethinylestradiol (EE2) and 17β-trenbolone (TB), on expressions of EDC-responsive genes in Japanese medaka (Oryzias latipes) at the cellular/tissue level paired with histological observation. Gene expressions of vitellogenin-II (Vit-II), androgen receptor (AR), and cytochrome P450 gonadal aromatase (CYP19a) were determined after exposure to 5, 50, or 500 ng/L of EE2 or 50, 500, or 5,000 ng/L of TB for 7 d. Exposure to the greatest concentration of EE2 or TB significantly reduced fecundity and caused histological alterations in gonads. 17α-Ethinylestradiol induced Vit-II expression in both male gonads and liver relative to controls and resulted in greater intensity of hematoxylin staining in hepatocytes, which was significantly correlated with Vit-II induction in liver. When exposed to EE2 at less than 50 ng/L, CYP19a expression associated with early stage oocytes was greater than that in controls. However, at 500 ng/L, this trend was reversed. The greater Vit-II expression in testis from all EE2 groups, and the lesser expression of CYP19a in ovaries from the 500 ng/L group, likely is related to changes in the number of cells in which these genes are predominantly expressed rather than to an increase in expression per cell. 17β-Trenbolone significantly induced AR expression in ovaries but did not alter AR expression in female liver. It was concluded that FISH combined with histology enables advanced elucidation of molecular effects of chemicals by associating changes in gene expression with certain tissues and/or cell types and allows these changes to be related to histological effects.

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