PUBLICATION

Generation of a fluorescent transgenic zebrafish for detection of environmental estrogens

Authors
Chen, H., Hu, J., Yang, J., Wang, Y., Xu, H., Jiang, Q., Gong, Y., Gu, Y., and Song, H.
ID
ZDB-PUB-091101-4
Date
2010
Source
Aquatic toxicology (Amsterdam, Netherlands)   96(1): 53-61 (Journal)
Registered Authors
Jiang, Qiu, Song, Houyan, Wang, Yuexiang, Xu, Hui
Keywords
Transgenic zebrafish, Environmental estrogens, Vitellogenin, Estrogen response element, Green fluorescent protein
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Base Sequence
  • Embryo, Nonmammalian
  • Endocrine Disruptors/pharmacology
  • Environmental Monitoring/methods*
  • Estrogens
  • Ethinyl Estradiol/analysis*
  • Ethinyl Estradiol/pharmacology
  • Female
  • Gene Expression Regulation/drug effects
  • Green Fluorescent Proteins/genetics
  • Green Fluorescent Proteins/metabolism*
  • Larva
  • Liver/metabolism
  • Male
  • Molecular Sequence Data
  • Sensitivity and Specificity
  • Vitellogenins/genetics
  • Water Pollutants, Chemical/analysis*
  • Water Pollutants, Chemical/pharmacology
  • Zebrafish/genetics*
  • Zebrafish/metabolism*
  • Zebrafish Proteins/genetics
PubMed
19850364 Full text @ Aquat. Toxicol.
Abstract
To establish a novel in vivo test system for rapid detection of environmental estrogens, an ere-zvtg1: gfp transgenic zebrafish line has been generated. In this transgenic line, under control conditions, GFP was exclusively expressed in the liver of mature adult female fish. Male and larval transgenic fish did not express GFP but could be induced to express GFP in the liver after exposure to 17-alpha-ethynylestradiol (EE(2)). Concurrent accumulation of zvtg1 and gfp mRNAs in embryos and larvae after EE(2) exposure was observed, which indicated that the expression of gfp transgene was driven by the zvtg1 promoter. Green fluorescence was first observed in the liver at 53, 74, 100 or 131h post-fertilization (hpf) after exposure to 100, 10, 1 or 0.1ng/L EE(2) from 1 to 2 cell stage, respectively. As for mature male transgenic zebrafish, green fluorescence was observed after exposure to 100, 10, 1 or 0.1ng/L EE(2) for 2, 3, 4 or 7 days, respectively; as for mature female, fluorescence was increased after exposure to relatively high concentrations of EE(2) (10 and 100ng/L). Green fluorescence in the liver was increased with prolonging of exposure time and was repeatedly induced after removal and re-addition of EE(2). We also demonstrated that GFP expression could be induced by other estrogenic compounds, including beta-estradiol (E(2), 0.1mug/L), cadmium chloride (CdCl(2), 10mug/L), zearalenone (50mug/L), estriol (E(3), 1mug/L), diethylstilbestrol (DES, 50ng/L) bisphenol A (BPA, 1mg/L) but not by weakly estrogenic compounds such as nonylphenol (NP, up to 10mg/L), or non-estrogenic steroid hormones such as progesterone (up to 100mg/L) and 17-hydroxysteroid (up to 50mg/L). These data suggest the transgenic zebrafish is sensitive and specific for detection of estrogenic compounds. Because the observed-effect concentrations are as low as those of environment and the observed-effect exposure times are very short, this transgenic fish is a promising candidate system for monitoring environmental estrogens directly, rapidly and easily.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping