PUBLICATION

Development of a convenient in vivo hepatotoxin assay using a transgenic zebrafish line with liver-specific DsRed expression

Authors
Zhang, X., Li, C., Gong, Z.
ID
ZDB-PUB-140513-360
Date
2014
Source
PLoS One   9: e91874 (Journal)
Registered Authors
Gong, Zhiyuan
Keywords
Fatty liver, Zebrafish, Isoniazid, Toxicity, Arsenates, Embryos, Ethanol, Fatty acids
MeSH Terms
  • Acetaminophen/adverse effects
  • Animals
  • Animals, Genetically Modified/genetics
  • Aspirin/adverse effects
  • Hazardous Substances/pharmacology
  • Humans
  • Isoniazid/adverse effects
  • Liver/drug effects*
  • Liver/pathology
  • Luminescent Proteins/biosynthesis*
  • Luminescent Proteins/genetics
  • Phenylbutazone/adverse effects
  • Zebrafish/genetics*
PubMed
24626481 Full text @ PLoS One
Abstract
Previously we have developed a transgenic zebrafish line (LiPan) with liver-specific red fluorescent protein (DsRed) expression under the fabp10a promoter. Since red fluorescence in the liver greatly facilitates the observation of liver in live LiPan fry, we envision that the LiPan zebrafish may provide a useful tool in analyses of hepatotoxicity based on changes of liver red fluorescence intensity and size. In this study, we first tested four well-established hepatotoxins (acetaminophen, aspirin, isoniazid and phenylbutazone) in LiPan fry and demonstrated that these hepatotoxins could significantly reduce both liver red fluorescence and liver size in a dosage-dependent manner, thus the two measurable parameters could be used as indicators of hepatotoxicity. We then tested the LiPan fry with nine other chemicals including environmental toxicants and human drugs. Three (mefenamic acid, lindane, and arsenate) behave like hepatotoxins in reduction of liver red fluorescence, while three others (17β-estradiol, TCDD [2,3,7,8-tetrachlorodibenzo-p-dioxin] and NDMA [N-nitrosodimethylamine]) caused increase of liver red fluorescence and the liver size. Ethanol and two other chemicals, amoxicillin (antibiotics) and chlorphenamine (pain killer) did not resulted in significant changes of liver red fluorescence and liver size. By quantitative RT-PCR analysis, we found that the changes of red fluorescence intensity caused by different chemicals correlated to the changes of endogenous fabp10a RNA expression, indicating that the measured hepatotoxicity was related to fatty acid transportation and metabolism. Finally we tested a mixture of four hepatotoxins and observed a significant reduction of red fluorescence in the liver at concentrations below the lowest effective concentrations of individual hepatotoxins, suggesting that the transgenic zebrafish assay is capable of reporting compound hepatotoxicity effect from chemical mixtures. Thus, the LiPan transgenic fry provide a rapid and convenient in vivo hepatotoxicity assay that should be applicable to high-throughput hepatotoxicity test in drug screening as well as in biomonitoring environmental toxicants.
Genes / Markers
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Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping