title: Effect-directed analysis for estrogenic compounds in a fluvial sediment sample using transgenic cyp19a1b-GFP zebrafish embryos.
abstract: Xenoestrogens may persist in the environment by binding to sediments or suspended particulate matter serving as long-term reservoir and source of exposure, particularly for organisms living in or in contact with sediments. In this study, we present for the first time an effect-directed analysis (EDA) for identifying estrogenic compounds in a sediment sample using embryos of a transgenic reporter fish strain. In the tg(cyp19a1b-GFP) transgenic zebrafish strain, the expression of GFP (green fluorescent protein) in the brain is driven by an oestrogen responsive element in the promoter of the cyp19a1b (aromatase) gene. The selected sediment sample of the Czech river Bilina had already been analysed in a previous EDA using the yeast oestrogen screening assay and had revealed fractions containing estrogenic compounds. When normal phase HPLC (high performance liquid chromatography) fractionation was used for the separation of the sediment sample, the biotest with transgenic fish embryos revealed two estrogenic fractions. Chemical analysis of candidate compounds in these sediment fractions suggested alkylphenols and estrone as candidate compounds responsible for the observed estrogenic effect. Alkylphenol concentrations could partially explain the estrogenicity of the fractions. However, xenoestrogens below the analytical detection limit or non-targeted estrogenic compounds have probably also contributed to the sample's estrogenic potency. The results indicated the suitability of the tg(cyp19a1b-GFP) fish embryo for an integrated chemical-biological analysis of estrogenic effects.
authors.affiliation: Department of Bioanalytical Ecotoxicology, Helmholtz Centre for Environmental Research - UFZ, Permoserstraße 15, 04318 Leipzig, Germany. Electronic address: eva.fetter@ufz.de.
authors.forename: Eva
authors.initials: E
authors.lastname: Fetter
authors.affiliation: Department of Effect-Directed Analysis, Helmholtz Centre for Environmental Research - UFZ, Permoserstraße 15, 04318 Leipzig, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Krauss
authors.affiliation: Unité d'écotoxicologie in vitro et in vivo, Direction des Risques Chroniques, Institut National de l'Environnement Industriel et des Risques (INERIS), BP 2, F-60550 Verneuil-en-Halatte, France.
authors.forename: François
authors.initials: F
authors.lastname: Brion
authors.affiliation: Research Institute for Environment, Health and Occupation, INSERM U1085, Biosit, Université de Rennes 1, Campus de Beaulieu, 35042 Rennes cedex, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Kah
authors.affiliation: Department of Bioanalytical Ecotoxicology, Helmholtz Centre for Environmental Research - UFZ, Permoserstraße 15, 04318 Leipzig, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Scholz
authors.affiliation: Department of Effect-Directed Analysis, Helmholtz Centre for Environmental Research - UFZ, Permoserstraße 15, 04318 Leipzig, Germany.
authors.forename: Werner
authors.initials: W
authors.lastname: Brack
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001141:Aromatase / Q000378:metabolism; D001681:Biological Assay; D004625:Embryo, Nonmammalian; D004784:Environmental Monitoring / Q000379:methods*; D004967:Estrogens / Q000032:analysis* / Q000378:metabolism; D004970:Estrone / Q000032:analysis / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D019015:Geologic Sediments / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D045483:Rivers / Q000737:chemistry; D014874:Water Pollutants, Chemical / Q000032:analysis* / Q000633:toxicity; D015027:Zebrafish / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D004967:Estrogens; D014874:Water Pollutants, Chemical; D029961:Zebrafish Proteins; D049452:Green Fluorescent Proteins; D004970:Estrone; D001141:Aromatase
keywords: Animal alternatives; Aromatase; Danio rerio; Endocrine disruptors; Environmental xenoestrogens; Fractionation