title: 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.
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.
authors.affiliation: CAS Key Lab of Bio-Medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, Jiangsu 215163, PR China.
authors.forename: Jingjing
authors.initials: J
authors.lastname: Tian
authors.affiliation: School of Biology & Basic Medical Sciences, Medical College, Soochow University, Suzhou 215123, Jiangsu, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Hu
authors.affiliation: CAS Key Lab of Bio-Medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, Jiangsu 215163, PR China.
authors.forename: Mingli
authors.initials: M
authors.lastname: Chen
authors.affiliation: CAS Key Lab of Bio-Medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, Jiangsu 215163, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China.
authors.forename: Huancai
authors.initials: H
authors.lastname: Yin
authors.affiliation: CAS Key Lab of Bio-Medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, Jiangsu 215163, PR China.
authors.forename: Peng
authors.initials: P
authors.lastname: Miao
authors.affiliation: CAS Key Lab of Bio-Medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, Jiangsu 215163, PR China.
authors.forename: Pengli
authors.initials: P
authors.lastname: Bai
authors.affiliation: CAS Key Lab of Bio-Medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, Jiangsu 215163, PR China. Electronic address: yinj@sibet.ac.cn.
authors.forename: Jian
authors.initials: J
authors.lastname: Yin
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D001564:Benzo(a)pyrene / Q000633:toxicity*; D064113:CRISPR-Cas Systems / Q000235:genetics; D019256:Cadmium Chloride / Q000633:toxicity*; D004625:Embryo, Nonmammalian / Q000187:drug effects / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism; D016296:Mutagenesis / Q000235:genetics; D011422:Propionates / Q000494:pharmacology; D021381:Protein Transport / Q000187:drug effects; D011804:Quinolines / Q000494:pharmacology; D018675:Toxicity Tests; D015027:Zebrafish / Q000196:embryology* / Q000378:metabolism*; D029961:Zebrafish Proteins / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism
chemical_list: D001564:Benzo(a)pyrene; D019256:Cadmium Chloride; D005978:Glutathione; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D011422:Propionates; D011804:Quinolines; D029961:Zebrafish Proteins; C000619860:multi-resistance associated protein 1, zebrafish; C059141:verlukast
keywords: Benzo[a]pyrene; CRISPR/Cas9; Cadmium chloride; Multi-resistance associated protein 1; Zebrafish embryo