title: Integrated biomarkers induced by chlorpyrifos in two different life stages of zebrafish (Danio rerio) for environmental risk assessment.
abstract: This study was performed to understand how chlorpyrifos (CHL) affects zebrafish (Danio rerio) embryos and adults, by exposing this model organism to various concentrations of the insecticide. The 96-h acute toxicity test to determine the effect of CHL on adult zebrafish yielded a LC50 of 709.43μg/L(-1). Small molecular weight proteins less than 25kDa and phospholipids were analyzed with MALDI-TOF MS/MS in order to compare expression patterns, revealing that some peaks were dramatically altered after CHL treatment. Whereas no acute toxicity was detected in the embryo toxicity test, malformation of zebrafish larvae was observed, with many individuals harboring curved spines. In an angiogenesis test on larvae of transgenic zebrafish, CHL did not have an inhibitory effect. Relative gene expression analyses using real-time polymerase chain reaction (RT-PCR) of DNA from zebrafish embryos revealed that different subtypes of cytochrome P450 (CYP450), such as CYP1A and CYP3A, were significantly up-regulated in response to CHL at a concentration of 400μg/L(-1) compared to the control. The expression level of NR1I2, a CYP gene transcriptional regulator, UGT1a1, and MDR1 were all up-regulated in the CHL-treated embryos. Finally, the expression level of acetylcholinesterase (AChE) and catalase (CAT) decreased, whereas that of superoxide dismutase (SOD) did not differ significantly. Our results suggest that the up-regulation of metabolic enzymes including CYP450 and MDR1 may be involved in CHL resistance in zebrafish.
authors.affiliation: School of Applied Biosciences, Kyungpook National University, Daegu 702-701, Republic of Korea.
authors.forename: Hwang-Ju
authors.initials: HJ
authors.lastname: Jeon
authors.affiliation: Institute of Ecological Phytochemistry, Department of Plant Life & Environmental Science, Hankyong National University, Anseong 456-749, Republic of Korea.
authors.forename: Yong-Ho
authors.initials: YH
authors.lastname: Lee
authors.affiliation: School of Life Science and Biotechnology, Kyungpook National University, Daegu 702-701, Republic of Korea.
authors.forename: Myoung-Jin
authors.initials: MJ
authors.lastname: Kim
authors.affiliation: School of Urban and Environmental Engineering, Ulsan National Institute of Science and Technology, Ulsan 44919, Republic of Korea.
authors.forename: Sung-Deuk
authors.initials: SD
authors.lastname: Choi
authors.affiliation: Chemical Safety Division, National Academy of Agricultural Science, Rural Development Administration, Jeonju 54875, Republic of Korea.
authors.forename: Byung-Jun
authors.initials: BJ
authors.lastname: Park
authors.affiliation: School of Applied Biosciences, Kyungpook National University, Daegu 702-701, Republic of Korea. Electronic address: selpest@knu.ac.kr.
authors.forename: Sung-Eun
authors.initials: SE
authors.lastname: Lee
mesh_terms: D000110:Acetylcholinesterase / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002374:Catalase / Q000378:metabolism; D004390:Chlorpyrifos / Q000633:toxicity*; D004625:Embryo, Nonmammalian; D004784:Environmental Monitoring; D007306:Insecticides / Q000633:toxicity*; D060888:Real-Time Polymerase Chain Reaction; D013482:Superoxide Dismutase / Q000378:metabolism; D014874:Water Pollutants, Chemical / Q000633:toxicity*; D015027:Zebrafish / Q000196:embryology / Q000502:physiology
chemical_list: D015415:Biomarkers; D007306:Insecticides; D014874:Water Pollutants, Chemical; D002374:Catalase; D013482:Superoxide Dismutase; D000110:Acetylcholinesterase; D004390:Chlorpyrifos
keywords: Biomarkers; CYP3A; Chlorpyrifos; MALDI-TOF MS/MS; MDR1; Zebrafish