PUBLICATION
A rapid in vivo zebrafish model to elucidate oxidative stress mediated PCB126-induced apoptosis and developmental toxicity
- Authors
- Liu, H., Gooneratne, R., Huang, X., Lai, R., Wei, J., Wang, W.
- ID
- ZDB-PUB-150317-11
- Date
- 2015
- Source
- Free radical biology & medicine 84: 91-102 (Journal)
- Registered Authors
- Keywords
- Apoptosis, Developmental toxicity, Glutathione metabolism, Nrf2a, PCB126, Zebrafish
- MeSH Terms
-
- Animals
- Antioxidants/pharmacology
- Apoptosis*
- Cystine/analogs & derivatives
- Cystine/pharmacology
- Environmental Pollutants/toxicity*
- Gene Expression Regulation, Developmental/drug effects
- Genes, Reporter
- Glutathione/metabolism
- Green Fluorescent Proteins/biosynthesis
- Green Fluorescent Proteins/genetics
- NF-E2-Related Factor 2/biosynthesis
- NF-E2-Related Factor 2/genetics
- Oxidative Stress*
- Polychlorinated Biphenyls/toxicity*
- Promoter Regions, Genetic
- Reactive Oxygen Species/metabolism
- Sequence Analysis, DNA
- Zebrafish
- Zebrafish Proteins/biosynthesis
- Zebrafish Proteins/genetics
- PubMed
- 25770664 Full text @ Free Radic. Biol. Med.
Citation
Liu, H., Gooneratne, R., Huang, X., Lai, R., Wei, J., Wang, W. (2015) A rapid in vivo zebrafish model to elucidate oxidative stress mediated PCB126-induced apoptosis and developmental toxicity. Free radical biology & medicine. 84:91-102.
Abstract
Dioxin-like 3,3',4,4',5-pentachlorobiphenyl (PCB126) is one of the most potent and widespread environmental pollutants. Although PCB126-induced toxicity is related to aryl hydrocarbon receptor (AHR) pathway, there is still no study that has constructed an in vivo visual model to clarify the role of the NRF2/ARE signaling pathway in oxidative stress mechanism of PCB126-induced toxicity. In the present study, an in vivo zebrafish model of nrf2a fused to enhanced green fluorescent protein (nrf2a-eGFP) was constructed. The zebrafish embryos microinjected with nrf2a-eGFP [72h post-fertilization (hpf)] were exposed to different concentrations of PCB126 [0, 25, 50, 100, 200μg/L], and 30mMN-acetylcysteine (NAC)+200μg/L PCB126. After 72h exposure, PCB126 significantly increased the malformation rates and induced the eGFP expression in a dose-dependent manner in several zebrafish tissue types. The distribution of eGFP fluorescence coincided with developmental deformity sites. NAC pre-treatment effectively counteracted PCB126 induced developmental toxicity including heart rate, pericardial edema and body length. Highest PCB126 dose 200μg/L produced marked apoptosis in the eye, gill and trunk detected by the terminal deoxynucleotide transferase-mediated dUTP nick-end labeling (TUNEL) assay. At 48 and 72h exposure, 200μg/L PCB126 affected glutathione metabolism as evidenced by decreased glutathione (GSH) and increased glutathione disulfide (GSSG) concentrations, indicative of oxidative stress. These effects were also counteracted by NAC pre-treatment. Furthermore, the Nrf2-regulated genes gclc, gpx, gstp1 and hmox1 were significantly induced at 24, 48 and 72h at the highest PCB126 exposures but not in the NAC pretreated group. In addition, a significant increase in ROS generation was detected in zebrafish larvae at 72h PCB126 exposure, which might offer a link for future mechanistic studies. Collectively, these data suggest that PCB126 induced developmental toxicity and apoptosis in nrf2a-eGFP injected zebrafish model are due to oxidative stress mediated by disruption to glutathione metabolism and changes in Nrf2-regulated gene expression.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping