Effects of combined stressors to cadmium and high temperature on antioxidant defense, apoptotic cell death, and DNA methylation in zebrafish (Danio rerio) embryos.

Fish are frequently affected by environmental stressors, such as temperature changes and heavy metal exposure, in aquatic ecosystems. In this study, we evaluated the combined effects of cadmium (Cd) toxicity and temperature (rearing temperature of 26 °C and heat stress at 34 °C) on zebrafish (Danio rerio) embryos. The survival and heart rates of zebrafish embryos decreased at relatively high Cd concentrations of 0.07 and 0.1 mg L. Abnormal morphology was induced by exposure to a combination of Cd toxicity and heat stress. The yolk sac edema size was not significantly different between the control- and Cd-treated groups. Cd exposure induced reactive oxygen species (ROS) production and cell death in the live zebrafish. High temperature (34 °C) triggered Cd-induced cell death and intracellular ROS production to a greater extent than the rearing temperature of 26 °C. Transcriptional levels of six genes-CAT, SOD, p53, BAX, Dnmt1, and Dnmt3b-were investigated. The mRNA expression of CAT and SOD, molecular indicators of oxidative stress, was increased significantly at 34 °C after Cd exposure. The mRNA expression of CAT was more sensitive to temperature than that of SOD in Cd-treated zebrafish. p53 and BAX, apoptosis-related genes, were upregulated upon combined exposure to high temperature and Cd. In addition, at 34 °C, the expression of Dnmt1 and Dnmt3b transcripts, markers of DNA methylation, was increased upon exposure of zebrafish to all concentrations of Cd. Overall, these results suggest that high temperature facilitates the potential role of Cd toxicity in the transcriptional regulation of genes involved in the antioxidant system, apoptosis, and DNA methylation.