PUBLICATION

The use of mrp1-deficient (Danio rerio) zebrafish embryos to investigate the role of Mrp1 in the toxicity of cadmium chloride and benzo[a]pyrene

Authors
Tian, J., Hu, J., Chen, M., Yin, H., Miao, P., Bai, P., Yin, J.
ID
ZDB-PUB-170311-1
Date
2017
Source
Aquatic toxicology (Amsterdam, Netherlands)   186: 123-133 (Journal)
Registered Authors
Hu, Jia, Tian, Jingjing, Yin, Jian
Keywords
Benzo[a]pyrene, CRISPR/Cas9, Cadmium chloride, Multi-resistance associated protein 1, Zebrafish embryo
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Base Sequence
  • Benzo(a)pyrene/toxicity*
  • CRISPR-Cas Systems/genetics
  • Cadmium Chloride/toxicity*
  • Embryo, Nonmammalian/drug effects
  • Embryo, Nonmammalian/metabolism*
  • Glutathione/metabolism
  • Multidrug Resistance-Associated Proteins/deficiency*
  • Multidrug Resistance-Associated Proteins/genetics
  • Multidrug Resistance-Associated Proteins/metabolism
  • Mutagenesis/genetics
  • Propionates/pharmacology
  • Protein Transport/drug effects
  • Quinolines/pharmacology
  • Toxicity Tests*
  • Zebrafish/embryology*
  • Zebrafish/metabolism*
  • Zebrafish Proteins/deficiency*
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism
PubMed
28282619 Full text @ Aquat. Toxicol.
Abstract
Previous studies in our lab have revealed that both P-glycoprotein (Pgp) and multi-resistance associated protein (Mrp) 1 played important roles in the detoxification of heavy metals and polycyclic aromatic hydrocarbon (PAH) in zebrafish embryos. This paper aims to extend this research by using mrp1-deficient model to illustrate the individual function of Mrp1. In this respect, CRISPR/Cas9 system was employed to generate a frameshift mutation in zebrafish mrp1 causing premature translational stops in Mrp1. Significant reduction on the efflux function of Mrps was found in mutant zebrafish embryos, which correlated well with the significantly enhanced accumulation and toxicity of cadmium chloride (CdCl2) and benzo[a]pyrene (BαP), indicating the protective role of the corresponding protein. The different alteration on the accumulation and toxicity of Cd2+ and BαP could be attributed to the fact that Cd2+ and its metabolites were mainly excreted by Mrp1, while BαP was primarily pumped out by Pgp. More importantly, the compensation mechanism for the absence of Mrp1, including elevated glutathione (GSH) level and up-regulated expression of pgp and mrp2 were also found. Thus, mrp1-deficient zebrafish embryo could be a useful tool in the investigation of Mrp1 functions in the early life stages of aquatic organisms. However, compensation mechanism should be taken into consideration in the interpretation of results obtained with mrp1-deficient fish.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping