From mRNA Expression of Drug Disposition Genes to In Vivo Assessment of CYP-Mediated Biotransformation during Zebrafish Embryonic and Larval Development

• Main Authors: Evy Verbueken, Chloé Bars, Jonathan S. Ball, Jelena Periz-Stanacev, Waleed F. A. Marei, Anna Tochwin, Isabelle J. Gabriëls, Ellen D. G. Michiels, Evelyn Stinckens, Lucia Vergauwen, Dries Knapen, Chris J. Van Ginneken, Steven J. Van Cruchten
• Format: Article
• Language: English
• Published: MDPI AG 2018-12-01
• Series: International Journal of Molecular Sciences
• Subjects: zebrafish; embryo; larva; development; cytochrome P450; phase II; drug transporter; biotransformation; activity; and expression
• Online Access: https://www.mdpi.com/1422-0067/19/12/3976

The zebrafish (<i>Danio rerio</i>) embryo is currently explored as an alternative for developmental toxicity testing. As maternal metabolism is lacking in this model, knowledge of the disposition of xenobiotics during zebrafish organogenesis is pivotal in order to correctly interpret the outcome of teratogenicity assays. Therefore, the aim of this study was to assess cytochrome P450 (CYP) activity in zebrafish embryos and larvae until 14 d post-fertilization (dpf) by using a non-specific CYP substrate, i.e., benzyloxy-methyl-resorufin (BOMR) and a CYP1-specific substrate, i.e., 7-ethoxyresorufin (ER). Moreover, the constitutive mRNA expression of <i>CYP1A</i>, <i>CYP1B1</i>, <i>CYP1C1</i>, <i>CYP1C2</i>, <i>CYP2K6</i>, <i>CYP3A65</i>, <i>CYP3C1</i>, phase II enzymes uridine diphosphate glucuronosyltransferase 1A1 (<i>UGT1A1</i>) and sulfotransferase 1st1 (<i>SULT1ST1</i>), and an ATP-binding cassette (ABC) drug transporter, i.e., <i>abcb4</i>, was assessed during zebrafish development until 32 dpf by means of quantitative PCR (qPCR). The present study showed that trancripts and/or the activity of these proteins involved in disposition of xenobiotics are generally low to undetectable before 72 h post-fertilization (hpf), which has to be taken into account in teratogenicity testing. Full capacity appears to be reached by the end of organogenesis (i.e., 120 hpf), although <i>CYP1</i>&#8212;except <i>CYP1A</i>&#8212;and <i>SULT1ST1</i> were shown to be already mature in early embryonic development.