In Vitro Biotransformation of Two Human CYP3A Probe Substrates and Their Inhibition during Early Zebrafish Development
At present, the zebrafish embryo is increasingly used as an alternative animal model to screen for developmental toxicity after exposure to xenobiotics. Since zebrafish embryos depend on their own drug-metabolizing capacity, knowledge of their intrinsic biotransformation is pivotal in order to corre...
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| Format: | Article |
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MDPI AG
2017-01-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | http://www.mdpi.com/1422-0067/18/1/217 |
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| author | Evy Verbueken Derek Alsop Moayad A. Saad Casper Pype Els M. Van Peer Christophe R. Casteleyn Chris J. Van Ginneken Joanna Wilson Steven J. Van Cruchten |
| author_facet | Evy Verbueken Derek Alsop Moayad A. Saad Casper Pype Els M. Van Peer Christophe R. Casteleyn Chris J. Van Ginneken Joanna Wilson Steven J. Van Cruchten |
| author_sort | Evy Verbueken |
| collection | DOAJ |
| description | At present, the zebrafish embryo is increasingly used as an alternative animal model to screen for developmental toxicity after exposure to xenobiotics. Since zebrafish embryos depend on their own drug-metabolizing capacity, knowledge of their intrinsic biotransformation is pivotal in order to correctly interpret the outcome of teratogenicity assays. Therefore, the aim of this in vitro study was to assess the activity of cytochrome P450 (CYP)—a group of drug-metabolizing enzymes—in microsomes from whole zebrafish embryos (ZEM) of 5, 24, 48, 72, 96 and 120 h post-fertilization (hpf) by means of a mammalian CYP substrate, i.e., benzyloxy-methyl-resorufin (BOMR). The same CYP activity assays were performed in adult zebrafish liver microsomes (ZLM) to serve as a reference for the embryos. In addition, activity assays with the human CYP3A4-specific Luciferin isopropyl acetal (Luciferin-IPA) as well as inhibition studies with ketoconazole and CYP3cide were carried out to identify CYP activity in ZLM. In the present study, biotransformation of BOMR was detected at 72 and 96 hpf; however, metabolite formation was low compared with ZLM. Furthermore, Luciferin-IPA was not metabolized by the zebrafish. In conclusion, the capacity of intrinsic biotransformation in zebrafish embryos appears to be lacking during a major part of organogenesis. |
| first_indexed | 2024-04-13T05:35:35Z |
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| institution | Directory Open Access Journal |
| issn | 1422-0067 |
| language | English |
| last_indexed | 2024-04-13T05:35:35Z |
| publishDate | 2017-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-36ee52e512734524bf15c833acd3f60d2022-12-22T03:00:17ZengMDPI AGInternational Journal of Molecular Sciences1422-00672017-01-0118121710.3390/ijms18010217ijms18010217In Vitro Biotransformation of Two Human CYP3A Probe Substrates and Their Inhibition during Early Zebrafish DevelopmentEvy Verbueken0Derek Alsop1Moayad A. Saad2Casper Pype3Els M. Van Peer4Christophe R. Casteleyn5Chris J. Van Ginneken6Joanna Wilson7Steven J. Van Cruchten8Applied Veterinary Morphology, Department of Veterinary Sciences, University of Antwerp, 2610 Wilrijk, Antwerp, BelgiumWilson Tox Lab, Department of Biology, McMaster University, 1280 Main Street West, Hamilton, ON L8S 4K1, CanadaApplied Veterinary Morphology, Department of Veterinary Sciences, University of Antwerp, 2610 Wilrijk, Antwerp, BelgiumApplied Veterinary Morphology, Department of Veterinary Sciences, University of Antwerp, 2610 Wilrijk, Antwerp, BelgiumApplied Veterinary Morphology, Department of Veterinary Sciences, University of Antwerp, 2610 Wilrijk, Antwerp, BelgiumApplied Veterinary Morphology, Department of Veterinary Sciences, University of Antwerp, 2610 Wilrijk, Antwerp, BelgiumApplied Veterinary Morphology, Department of Veterinary Sciences, University of Antwerp, 2610 Wilrijk, Antwerp, BelgiumWilson Tox Lab, Department of Biology, McMaster University, 1280 Main Street West, Hamilton, ON L8S 4K1, CanadaApplied Veterinary Morphology, Department of Veterinary Sciences, University of Antwerp, 2610 Wilrijk, Antwerp, BelgiumAt present, the zebrafish embryo is increasingly used as an alternative animal model to screen for developmental toxicity after exposure to xenobiotics. Since zebrafish embryos depend on their own drug-metabolizing capacity, knowledge of their intrinsic biotransformation is pivotal in order to correctly interpret the outcome of teratogenicity assays. Therefore, the aim of this in vitro study was to assess the activity of cytochrome P450 (CYP)—a group of drug-metabolizing enzymes—in microsomes from whole zebrafish embryos (ZEM) of 5, 24, 48, 72, 96 and 120 h post-fertilization (hpf) by means of a mammalian CYP substrate, i.e., benzyloxy-methyl-resorufin (BOMR). The same CYP activity assays were performed in adult zebrafish liver microsomes (ZLM) to serve as a reference for the embryos. In addition, activity assays with the human CYP3A4-specific Luciferin isopropyl acetal (Luciferin-IPA) as well as inhibition studies with ketoconazole and CYP3cide were carried out to identify CYP activity in ZLM. In the present study, biotransformation of BOMR was detected at 72 and 96 hpf; however, metabolite formation was low compared with ZLM. Furthermore, Luciferin-IPA was not metabolized by the zebrafish. In conclusion, the capacity of intrinsic biotransformation in zebrafish embryos appears to be lacking during a major part of organogenesis.http://www.mdpi.com/1422-0067/18/1/217zebrafishembryodevelopmentcytochrome P450activityin vitroand biotransformation |
| spellingShingle | Evy Verbueken Derek Alsop Moayad A. Saad Casper Pype Els M. Van Peer Christophe R. Casteleyn Chris J. Van Ginneken Joanna Wilson Steven J. Van Cruchten In Vitro Biotransformation of Two Human CYP3A Probe Substrates and Their Inhibition during Early Zebrafish Development International Journal of Molecular Sciences zebrafish embryo development cytochrome P450 activity in vitro and biotransformation |
| title | In Vitro Biotransformation of Two Human CYP3A Probe Substrates and Their Inhibition during Early Zebrafish Development |
| title_full | In Vitro Biotransformation of Two Human CYP3A Probe Substrates and Their Inhibition during Early Zebrafish Development |
| title_fullStr | In Vitro Biotransformation of Two Human CYP3A Probe Substrates and Their Inhibition during Early Zebrafish Development |
| title_full_unstemmed | In Vitro Biotransformation of Two Human CYP3A Probe Substrates and Their Inhibition during Early Zebrafish Development |
| title_short | In Vitro Biotransformation of Two Human CYP3A Probe Substrates and Their Inhibition during Early Zebrafish Development |
| title_sort | in vitro biotransformation of two human cyp3a probe substrates and their inhibition during early zebrafish development |
| topic | zebrafish embryo development cytochrome P450 activity in vitro and biotransformation |
| url | http://www.mdpi.com/1422-0067/18/1/217 |
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