Embryonic Zebrafish Danio Rerio Research Articles

Toxicity of micro and nano tire particles and leachate for model freshwater organisms

Environmental sampling has documented a diversity of microplastics, including high levels of black rubber— generally identified as tire debris. Though organisms have been shown to ingest tire particles (TPs), past research focused on toxicity of leachate alone, overlooking potential effects of particles. To address these gaps, we assessed the toxicity of micro (1–20 µm) and nano (<1 µm) TPs for two model organisms, embryonic Zebrafish Danio rerio and the crustacean Daphnia magna. To assess effects on development, Zebrafish embryos were exposed to concentrations of TPs or leachate ranging from 0 to 3.0 × 109 particles/ml and 0–100% respectively (n = 4). Greater mortality and sublethal malformations were observed following nano TP and leachate exposures as compared to micro TPs. Unique abnormalities between the exposures indicates that there is both chemical and particle-specific toxicity. We also observed D. magna mortality following a 48 h exposure of neonate to TPs or leachate, ranging from 0 to 3.3 × 109 particles/ml and 0–100% respectively (n = 3). Though, particle-enhancement of toxicity was observed for both Zebrafish and D. magna, overall sensitivity to TPs differed. It is important to identify differential toxicities across species to achieve an understanding of the environmental impacts of TPs and the chemicals they leach.

Specific RNAi Mediated Gene Knockdown in Zebrafish Cell Lines

Here we demonstrate highly efficient RNA interference in ZFL, SJD and ZF4 cell lines derived from adult and embryonic zebrafish Danio rerio. Microinjection of siRNAs resulted in silencing in almost 100% of cells while transfection using cationic liposomes led to silencing in 30%. Use of siRNAs against zebrafish lamin A, lamin B2 and the motor protein Eg5, led to knockdown of the target genes with the specific phenotypes expected from prior studies in mammalian cells. In contrast injection of lamin A, GL2 and eGFP siRNAs into zebrafish embryos resulted in morphological defects, abnormal development and early death of most embryos. The results indicate unspecific responses to siRNAs in the embryo but a fully developed and active RNAi machinery in cell lines.