Screening of the estrogenic activity of environmental samples using transgenic cyp19a1b-GFP zebrafish embryos

Abstract : Due to the potential environmental hazards on reproduction and population development in the aquatic ecosystem by estrogenic substances, their hazard assessment play a paramount role for regulation of chemicals, biocides, pesticides and pharmaceuticals in the EU and worldwide. Furthermore, they are also highly relevant for biomonitoring within the EU Water Framework Directive. For the assessment of their potential adverse effects on the environment, biological test systems play an essential role. The identification of endocrine disrupters with the chemical analysis is limited due to the low but still biologically relevant concentration in the surface waters. Besides, assessing the environmental hazard of mixtures of endocrine disrupters through classical monitoring is not only expensive but also not easily predictable. For this reason, effect-directed analysis using bioassays can be an excellent supplement of the classical monitoring of aquatic ecosystems. Transgenic models appear to be a powerful tool for high- throughput screening of changes in the gene expression induced by estrogenic chemicals. Using the zebrafishcyp19a1b-GFP transgenic embryo, which expresses GFP under the control of the zebrafish cyp19a1b promoter, adds the complexity of a full organism to the simplicity and reproducibility of cellular reporter gene assays. In this study we present the first results using this transgenic line for effect-directed analysis of a contaminated sediment sample. In this sample from the Czech river Bilina a previous analysis using the yeast screening assay has identified fractions containing estrogenic compounds. We were able to confirm this study by means of the transgeniccyp19a1b-GFP zebrafish strain. The results were also validated by artificial mixtures of compounds, which were identified by the effect-directed analysis, resembling the compositions of the fractions. Our study also includes data on the validation of the transgenic zebrafish strain with known estrogenic substances and its confirmation using molecular and an in-vitro method. The transgenic cyp19a1b-GFP in-vivo assay proved to be an excellent screening method with high sensitivity and reproducibility for environmental samples.
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Conference papers
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Submitted on : Wednesday, April 2, 2014 - 3:58:43 PM
Last modification on : Wednesday, January 23, 2019 - 10:42:03 AM

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  • HAL Id : ineris-00970972, version 1
  • INERIS : EN-2012-163

Citation

E. Fetter, S. Scholz, M. Krauss, Werner Brack, K. Petersen, et al.. Screening of the estrogenic activity of environmental samples using transgenic cyp19a1b-GFP zebrafish embryos. 6. SETAC World Congress 2012 / 22. SETAC Europe Annual Meeting "Securing a sustainable future : integrating science, policy and people", May 2012, Berlin, Germany. ⟨ineris-00970972⟩

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