Transgenic Zebrafish Expressing Rat Cytochrome P450 2E1 (CYP2E1): Augmentation of Acetaminophen-Induced Toxicity in the Liver and Retina
Metabolic activation is the primary cause of chemical toxicity including hepatotoxicity. Cytochrome P450 2E (CYP2E) is involved in this process for many hepatotoxicants, including acetaminophen (APAP), one of the most common analgesics and antipyretics. Although the zebrafish is now used as a model...
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MDPI AG
2023-02-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/4/4013 |
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| author | Yoshinori Sato Wenjing Dong Tatsuro Nakamura Naohiro Mizoguchi Tasuku Nawaji Miyu Nishikawa Takenori Onaga Shinichi Ikushiro Makoto Kobayashi Hiroki Teraoka |
| author_facet | Yoshinori Sato Wenjing Dong Tatsuro Nakamura Naohiro Mizoguchi Tasuku Nawaji Miyu Nishikawa Takenori Onaga Shinichi Ikushiro Makoto Kobayashi Hiroki Teraoka |
| author_sort | Yoshinori Sato |
| collection | DOAJ |
| description | Metabolic activation is the primary cause of chemical toxicity including hepatotoxicity. Cytochrome P450 2E (CYP2E) is involved in this process for many hepatotoxicants, including acetaminophen (APAP), one of the most common analgesics and antipyretics. Although the zebrafish is now used as a model for toxicology and toxicity tests, the CYP2E homologue in zebrafish has not been identified yet. In this study, we prepared transgenic zebrafish embryos/larvae expressing rat CYP2E1 and enhanced green fluorescent protein (EGFP) using a β-actin promoter. Rat CYP2E1 activity was confirmed by the fluorescence of 7-hydroxycoumarin (7-HC), a metabolite of 7-methoxycoumarin that was specific for CYP2 in transgenic larvae with EGFP fluorescence (EGFP [+]) but not in transgenic larvae without EGFP fluorescence (EGFP [−]). APAP (2.5 mM) caused reduction in the size of the retina in EGFP [+] larvae but not in EGFP [−] larvae, while APAP similarly reduced pigmentation in both larvae. APAP at even 1 mM reduced the liver size in EGFP [+] larvae but not in EGFP [−] larvae. APAP-induced reduction of liver size was inhibited by <i>N</i>-acetylcysteine. These results suggest that rat CYP2E1 is involved in some APAP-induced toxicological endpoints in the retina and liver but not in melanogenesis of the developing zebrafish. |
| first_indexed | 2024-03-11T08:41:02Z |
| format | Article |
| id | doaj.art-a1def183e21447fc905e76ded3565d45 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T08:41:02Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-a1def183e21447fc905e76ded3565d452023-11-16T21:08:16ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-02-01244401310.3390/ijms24044013Transgenic Zebrafish Expressing Rat Cytochrome P450 2E1 (CYP2E1): Augmentation of Acetaminophen-Induced Toxicity in the Liver and RetinaYoshinori Sato0Wenjing Dong1Tatsuro Nakamura2Naohiro Mizoguchi3Tasuku Nawaji4Miyu Nishikawa5Takenori Onaga6Shinichi Ikushiro7Makoto Kobayashi8Hiroki Teraoka9School of Veterinary Medicine, Rakuno Gakuen University, 582, Bunkyodai-Midorimachi, Ebetsu 069-8501, Hokkaido, JapanSchool of Veterinary Medicine, Rakuno Gakuen University, 582, Bunkyodai-Midorimachi, Ebetsu 069-8501, Hokkaido, JapanSchool of Veterinary Medicine, Rakuno Gakuen University, 582, Bunkyodai-Midorimachi, Ebetsu 069-8501, Hokkaido, JapanChemicals Evaluation and Research Institute, Japan (CERI), 3-2-7, Miyanojin, Kurume 839-0801, Fukuoka, JapanChemicals Evaluation and Research Institute, Japan (CERI), 3-2-7, Miyanojin, Kurume 839-0801, Fukuoka, JapanDepartment of Biotechnology, Faculty of Engineering, Toyama Prefectural University, 5180, Kurokawa, Imizu 939-0398, Toyama, JapanSchool of Veterinary Medicine, Rakuno Gakuen University, 582, Bunkyodai-Midorimachi, Ebetsu 069-8501, Hokkaido, JapanDepartment of Biotechnology, Faculty of Engineering, Toyama Prefectural University, 5180, Kurokawa, Imizu 939-0398, Toyama, JapanDepartment of Molecular and Developmental Biology, Institute of Medicine, University of Tsukuba, Tsukuba 305-8575, Ibaraki, JapanSchool of Veterinary Medicine, Rakuno Gakuen University, 582, Bunkyodai-Midorimachi, Ebetsu 069-8501, Hokkaido, JapanMetabolic activation is the primary cause of chemical toxicity including hepatotoxicity. Cytochrome P450 2E (CYP2E) is involved in this process for many hepatotoxicants, including acetaminophen (APAP), one of the most common analgesics and antipyretics. Although the zebrafish is now used as a model for toxicology and toxicity tests, the CYP2E homologue in zebrafish has not been identified yet. In this study, we prepared transgenic zebrafish embryos/larvae expressing rat CYP2E1 and enhanced green fluorescent protein (EGFP) using a β-actin promoter. Rat CYP2E1 activity was confirmed by the fluorescence of 7-hydroxycoumarin (7-HC), a metabolite of 7-methoxycoumarin that was specific for CYP2 in transgenic larvae with EGFP fluorescence (EGFP [+]) but not in transgenic larvae without EGFP fluorescence (EGFP [−]). APAP (2.5 mM) caused reduction in the size of the retina in EGFP [+] larvae but not in EGFP [−] larvae, while APAP similarly reduced pigmentation in both larvae. APAP at even 1 mM reduced the liver size in EGFP [+] larvae but not in EGFP [−] larvae. APAP-induced reduction of liver size was inhibited by <i>N</i>-acetylcysteine. These results suggest that rat CYP2E1 is involved in some APAP-induced toxicological endpoints in the retina and liver but not in melanogenesis of the developing zebrafish.https://www.mdpi.com/1422-0067/24/4/4013acetaminophenbioactivationCYP2E1ratzebrafish |
| spellingShingle | Yoshinori Sato Wenjing Dong Tatsuro Nakamura Naohiro Mizoguchi Tasuku Nawaji Miyu Nishikawa Takenori Onaga Shinichi Ikushiro Makoto Kobayashi Hiroki Teraoka Transgenic Zebrafish Expressing Rat Cytochrome P450 2E1 (CYP2E1): Augmentation of Acetaminophen-Induced Toxicity in the Liver and Retina International Journal of Molecular Sciences acetaminophen bioactivation CYP2E1 rat zebrafish |
| title | Transgenic Zebrafish Expressing Rat Cytochrome P450 2E1 (CYP2E1): Augmentation of Acetaminophen-Induced Toxicity in the Liver and Retina |
| title_full | Transgenic Zebrafish Expressing Rat Cytochrome P450 2E1 (CYP2E1): Augmentation of Acetaminophen-Induced Toxicity in the Liver and Retina |
| title_fullStr | Transgenic Zebrafish Expressing Rat Cytochrome P450 2E1 (CYP2E1): Augmentation of Acetaminophen-Induced Toxicity in the Liver and Retina |
| title_full_unstemmed | Transgenic Zebrafish Expressing Rat Cytochrome P450 2E1 (CYP2E1): Augmentation of Acetaminophen-Induced Toxicity in the Liver and Retina |
| title_short | Transgenic Zebrafish Expressing Rat Cytochrome P450 2E1 (CYP2E1): Augmentation of Acetaminophen-Induced Toxicity in the Liver and Retina |
| title_sort | transgenic zebrafish expressing rat cytochrome p450 2e1 cyp2e1 augmentation of acetaminophen induced toxicity in the liver and retina |
| topic | acetaminophen bioactivation CYP2E1 rat zebrafish |
| url | https://www.mdpi.com/1422-0067/24/4/4013 |
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