PUBLICATION
Increased susceptibility to oxidative stress-induced toxicological evaluation by genetically modified nrf2a-deficient zebrafish
- Authors
- Yamashita, A., Deguchi, J., Honda, Y., Yamada, T., Miyawaki, I., Nishimura, Y., Tanaka, T.
- ID
- ZDB-PUB-181231-1
- Date
- 2019
- Source
- Journal of Pharmacological and Toxicological Methods 96: 34-45 (Journal)
- Registered Authors
- Tanaka, Toshio
- Keywords
- Acetaminophen, Doxorubicin, Nrf2a-deficiency, Oxidative stress, Toxicity evaluation, Zebrafish
- MeSH Terms
-
- Acetaminophen/toxicity
- Animals
- Antioxidants/metabolism
- Chemical and Drug Induced Liver Injury/genetics
- Chemical and Drug Induced Liver Injury/metabolism
- Clustered Regularly Interspaced Short Palindromic Repeats
- Doxorubicin/toxicity
- Heart Function Tests/drug effects
- Hydrogen Peroxide/toxicity
- Larva/drug effects
- Larva/genetics
- Larva/metabolism
- Liver/metabolism
- Liver/pathology
- NF-E2-Related Factor 2/deficiency*
- NF-E2-Related Factor 2/genetics
- Oxidation-Reduction
- Oxidative Stress/drug effects*
- Oxidative Stress/genetics
- RNA, Messenger/metabolism
- Signal Transduction/drug effects
- Zebrafish/genetics*
- Zebrafish Proteins/deficiency*
- Zebrafish Proteins/genetics
- PubMed
- 30594530 Full text @ J. Pharmacol. Toxicol. Methods
Citation
Yamashita, A., Deguchi, J., Honda, Y., Yamada, T., Miyawaki, I., Nishimura, Y., Tanaka, T. (2019) Increased susceptibility to oxidative stress-induced toxicological evaluation by genetically modified nrf2a-deficient zebrafish. Journal of Pharmacological and Toxicological Methods. 96:34-45.
Abstract
Introduction Oxidative stress plays an important role in drug-induced toxicity. Oxidative stress-mediated toxicities can be detected using conventional animal models but their sensitivity is insufficient, and novel models to improve susceptibility to oxidative stress have been researched. In recent years, gene targeting methods in zebrafish have been developed, making it possible to generate homozygous null mutants. In this study, we established zebrafish deficient in the nuclear factor erythroid 2-related factor 2a (nrf2a), a key antioxidant-responsive gene, and its potential to detect oxidative stress-mediated toxicity was examined.
Methods Nrf2a-deficient zebrafish were generated using the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 technique. The loss of nrf2a function was confirmed by the tolerability to hydrogen peroxide and hydrogen peroxide-induced gene expression profiles being related to antioxidant response element (ARE)-dependent signaling. Subsequently, vulnerability of nrf2a-deficient zebrafish to acetaminophen (APAP)- or doxorubicin (DOX)-induced toxicity was investigated.
Results Nrf2a-deficient zebrafish showed higher mortality than wild type accompanied by less induction of ARE-dependent genes with hydrogen peroxide treatment. Subsequently, this model showed increased severity and incidence of APAP-induced hepatotoxicity or DOX-induced cardiotoxicity than wild type.
Discussion Our results demonstrated that anti-oxidative response might not fully function in this model, and resulted in higher sensitivity to drug-induced oxidative stress. Our data support the usefulness of nrf2a-deficient model as a tool for evaluation of oxidative stress-related toxicity in drug discovery research.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping