Zebrafish embryos as a screen for DNA methylation modifications after compound exposure

Manon C. Bouwmeester, Sander Ruiter, Tobias Lommelaars, Josefine Sippel, Hennie M. Hodemaekers, Evert-Jan van den Brandhof, Jeroen L.A. Pennings, Jorke H. Kamstra, Jaroslav Jelinek, Jean-Pierre J. Issa, Juliette Legler, Leo T.M. van der Ven

    Research output: Contribution to journalArticleAcademicpeer-review

    Abstract

    Modified epigenetic programming early in life is proposed to underlie the development of an adverse adult phenotype, known as the Developmental Origins of Health and Disease (DOHaD) concept. Several environmental contaminants have been implicated as modifying factors of the developing epigenome. This underlines the need to investigate this newly recognized toxicological risk and systematically screen for the epigenome modifying potential of compounds. In this study, we examined the applicability of the zebrafish embryo as a screening model for DNA methylation modifications. Embryos were exposed from 0 to 72h post fertilization (hpf) to bisphenol-A (BPA), diethylstilbestrol, 17α-ethynylestradiol, nickel, cadmium, tributyltin, arsenite, perfluoroctanoic acid, valproic acid, flusilazole, 5-azacytidine (5AC) in subtoxic concentrations. Both global and site-specific methylation was examined. Global methylation was only affected by 5AC. Genome wide locus-specific analysis was performed for BPA exposed embryos using Digital Restriction Enzyme Analysis of Methylation (DREAM), which showed minimal wide scale effects on the genome, whereas potential informative markers were not confirmed by pyrosequencing. Site-specific methylation was examined in the promoter regions of three selected genes vasa, vtg1 and cyp19a2, of which vasa (ddx4) was the most responsive. This analysis distinguished estrogenic compounds from metals by direction and sensitivity of the effect compared to embryotoxicity. In conclusion, the zebrafish embryo is a potential screening tool to examine DNA methylation modifications after xenobiotic exposure. The next step is to examine the adult phenotype of exposed embryos and to analyze molecular mechanisms that potentially link epigenetic effects and altered phenotypes, to support the DOHaD hypothesis.
    Original languageEnglish
    Pages (from-to)84-96
    Number of pages13
    JournalToxicology and Applied Pharmacology
    Volume291
    DOIs
    Publication statusPublished - 15 Jan 2016

    Keywords

    • DNA methylation
    • Environmental contaminant exposure
    • Epigenetics
    • Screening
    • Zebrafish embryo
    • 4,4' isopropylidenediphenol
    • arsenic trioxide
    • azacitidine
    • cadmium
    • diethylstilbestrol
    • ethinylestradiol
    • flusilazole
    • nickel
    • tributyltin
    • valproic acid
    • xenobiotic agent
    • adult
    • article
    • controlled study
    • DNA modification
    • embryo
    • embryotoxicity
    • environmental exposure
    • epigenetics
    • fertilization
    • gene locus
    • high resolution melting analysis
    • nonhuman
    • phenotype
    • promoter region
    • pyrosequencing
    • restriction mapping
    • zebra fish

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