Modification and quantification of in vivo EROD live-imaging with zebrafish (Danio rerio) embryos to detect both induction and inhibition of CYP1A

• Verfasst von: Kais, Britta [VerfasserIn]
• Verfasst von: Ottermanns, Richard [VerfasserIn]
• Verfasst von: Scheller, Franziska [VerfasserIn]
• Verfasst von: Braunbeck, Thomas [VerfasserIn]
• Titel: Modification and quantification of in vivo EROD live-imaging with zebrafish (Danio rerio) embryos to detect both induction and inhibition of CYP1A
• Verf.angabe: Britta Kais, Richard Ottermanns, Franziska Scheller, Thomas Braunbeck
• Umfang: 18 S.
• Fussnoten: Available online: 2 October 2017 ; Gesehen am 21.02.2018
• Titel Quelle: Enthalten in: The science of the total environment
• Jahr Quelle: 2018
• Band/Heft Quelle: 615(2018), S. 330-347
• ISSN Quelle: 1879-1026
• DOI: doi:10.1016/j.scitotenv.2017.09.257
• Datenträger: Online-Ressource
• Sprache: eng
• Bibliogr. Hinweis: Erscheint auch als Druck-Ausgabe: Kais, Britta: Modification and quantification of in vivo EROD live-imaging with zebrafish (Danio rerio) embryos to detect both induction and inhibition of CYP1A
• K10plus-PPN: 157007027X

Abstract

The visualization of specific activation of the aryl hydrocarbon receptor (AhR) directly in the zebrafish embryo (Danio rerio) via live-imaging is a reliable tool to investigate the presence of dioxin-like substances in environmental samples. The co-existence of inducers and inhibitors of cytochrome P450-dependent monooxygenases (CYP1A) is typical of complex environmental mixtures and requires modifications of the in vivo EROD assay: For this end, zebrafish embryos were used to evaluate the EROD-modifying potentials of common single-compound exposures as well as binary mixtures with the PAH-type Ah-receptor agonist β-naphthoflavone. For chemical testing, chlorpyrifos and Aroclor 1254 were selected; β-naphthoflavone served as maximum EROD induction control. Chlorpyrifos (≤EC10) could be documented to be a strong CYP1A inhibitor causing characteristic edema-related toxicity. Aroclor 1254 resulted in inhibition of CYP1A catalytic activity in a concentration- and specific time-dependent manner. Next to a fast CYP1A induction, CYP1A inhibition could also be detected after 3h short-term exposure of zebrafish embryos to chlorpyrifos. This communication also describes techniques for the quantification of fluorescence signals via densitometry as a basis for subsequent statistical assessment. The co-exposure approach with zebrafish embryos accounts for the nature of potential interaction between CYP1A inducers and inhibitors and thus pays tribute to the complexity of environmental mixtures. The co-exposure EROD live-imaging assay thus facilitates a better understanding of mixture effects and allows a better assessment and interpretation of (embryo) toxic potentials.