DNA methylation and gene expression alterations in zebrafish embryos exposed to cadmium.

An unexplored attributing molecular mechanism of Cd toxicity is interference with the epigenetic machinery, such as DNA methylation, processes that are crucial for early fetal development. In order to investigate the effects of Cd on the expression of metallothionein (MT) and Dnmts transcripts, markers of DNA methylation, and signaling pathway gene expression, zebrafish embryos were exposed during 24 hours post-fertilization (starting at maximum 8-cell stage) to 0.0089, 0.089, and 0.89 µM Cd. The results showed that the Cd accumulation in zebrafish embryo reached a stable level after 12 hpf, and the Cd accumulation at individual time points was significantly different among different concentration groups. MT mRNA fold was significantly positive with the Cd content in embryos. We observed that the expression level of DNA methyltransferase (Dnmts) in the 0.089 µM Cd exposure group was significantly up-regulated. Dnmt1 expression was significantly up-regulated in the 0.89 µM Cd exposure group, and Dnmt3s expression and global methylation levels were significantly down-regulated. Cd up-regulated ErbB-3 gene expression, down-regulated ErbB-4 gene expression, and neutralized ErbB-1 gene expression. Cd activated Ca2+, MAPK-JUK, p38 MAP kinase, PI3K-AKT, and VEGF signaling pathway genes, indicating these pathway genes related to Cd exposure level. The results are helpful to clarify the molecular mechanism of DNA methylation in zebrafish embryo under metal pressure and further interference with the epigenetic machinery.

Predicting the survival of zebrafish larvae exposed to fluctuating pulses of lead and cadmium.

Aquatic organisms are often exposed to time-varied concentrations of contaminants due to pulsed inputs in natural water. Traditional toxicology experiments are usually carried out in a constant exposure pattern, which is inconsistent with the actual environment. In this study, a refined toxicokinetic-toxicodynamic (TK-TD) model was used to study the toxic effects of Pb and Cd on zebrafish larvae under three pulse exposures with 2, 4, and 6 h, respectively. The parameter sensitivity analysis showed that JM, max had the greatest impact on the output of the model. Cd or Pb pulse exposure resulted in less death than constant exposure at the same time-weighted average (TWA) concentrations. Survival fraction in larvae under 6 h interval between two pulses of Pb or Cd was larger than that under 2 h and 4 h interval. Toxicity under constant exposure of Cd or Pb was greater than that under 2, 4, and 6 h interval pulse exposure because the cumulative Cd or Pb concentration in the body under constant exposure was greater than that under pulse exposure. The results also showed that the stochastic death (SD) model was more suitable than the individual tolerance (IT) model for predicting the survival fraction of larvae under pulse exposure to Pb and Cd, which was indicated by higher R2 (0.670-0.940) in SD model than that (0.588-0.861) in IT model. Our model provides approaches for laboratory toxicity testing and modeling approaches for addressing the toxicity of heavy metal pulsed exposure.