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
(all 24)
PubMed
30594530 Full text @ J. Pharmacol. Toxicol. Methods
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
Marker Marker Type Name
gclcGENEglutamate-cysteine ligase, catalytic subunit
gstp1.2GENEglutathione S-transferase pi 1.2
keap1aGENEkelch-like ECH-associated protein 1a
keap1bGENEkelch-like ECH-associated protein 1b
mitfaGENEmelanocyte inducing transcription factor a
nfe2l2aGENEnfe2 like bZIP transcription factor 2a
nfe2l2bGENEnfe2 like bZIP transcription factor 2b
prdx1GENEperoxiredoxin 1
txnbGENEthioredoxin b
1 - 9 of 9
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Figures
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Expression
Phenotype
Fish Conditions Stage Phenotype Figure
mitfab692/b692chemical treatment by environment: paracetamolDay 4
mitfab692/b692chemical treatment by environment: paracetamolDay 4
mitfab692/b692chemical treatment by environment: paracetamolDay 5
mitfab692/b692chemical treatment by environment: paracetamolDay 5
mitfab692/b692chemical treatment by environment: paracetamolDay 5
mitfab692/b692chemical treatment by environment: paracetamolDay 5
mitfab692/b692chemical treatment by environment: hydrogen peroxideDay 5
mitfab692/b692chemical treatment by environment: hydrogen peroxideDay 5
mitfab692/b692chemical treatment by environment: hydrogen peroxideDay 5
mitfab692/b692chemical treatment by environment: sulforaphane, chemical treatment by environment: hydrogen peroxideDay 5
1 - 10 of 26
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Mutations / Transgenics
Allele Construct Type Affected Genomic Region
b692
    		Point Mutation
    fh318
      		Point Mutation
      zf3103
        		Small Deletion
        1 - 3 of 3
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        Human Disease / Model
        No data available
        Sequence Targeting Reagents
        Target Reagent Reagent Type
        nfe2l2aCRISPR3-nfe2l2aCRISPR
        1 - 1 of 1
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        Fish
        Antibodies
        No data available
        Orthology
        Engineered Foreign Genes
        No data available
        Mapping
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        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.