Figure 2.

Schematic workflow for high throughput screening of different compounds in the zebrafish model. In a first step, zebrafish embryos with or without genetic manipulation via e.g., morpholino injection, are incubated in the presence of different substances (substances A to E in this example). The embryos are monitored to detect toxicity, developmental delay, and changes in phenotype. If the embryos develop phenotypically normal, fish are sacrificed and analyses of epigenetic changes are conducted, e.g., with bisulfite conversion of DNA with subsequent next-generation sequencing (NGS) or mass spectrometry (MS). Further evaluations include chromatin immune precipitation (ChIP) and quantitative PCR (qPCR). If significant results regarding the effect of the specific substance are obtained, the substance is tested further in a mammalian, e.g., murine, model.