Analysis of vascular disruption in zebrafish embryos as an endpoint to predict developmental toxicity - High Content Screening Raw Data (OBI/WIK strain)

A novel automated imaging-based method to detect inhibition of angiogenesis in early life stage zebrafish. Video subtraction was used to identify the location and number of functional intersegmental vessels according to the detection of moving blood cells. By exposing embryos to multiple tyrosine kinase inhibitors including SU4312, SU5416, Sorafenib, or PTK787, we confirmed that this method can detect concentration-dependent inhibition of angiogenesis. The new test method showed higher sensitivity, i.e. lower effect concentrations, relative to a fluorescent reporter gene strain (Tg(KDR:EGFP)) exposed to the same tyrosine kinase inhibitors. Indicating that functional effects due to altered tubulogenesis or blood transport can be detected before structural changes of the endothelium are visible by fluorescence imaging. Comparison of exposure windows indicated higher specificity for angiogenesis when exposure started at later embryonic stages (24 hours post fertilization). Our findings establish video imaging in wild-type strains as viable, non-invasive, high-throughput method for the detection of chemical-induced angiogenic disruption in zebrafish embryos. We established a lable-free and non-invasive method to visualize the blood vessel network in the developing wild type zebrafish embryo. We quantified ISV inhibition by comparing WT control, treated with tyrosine kinase inhibitors (SU4312, SU5416, Sorafenib and PTK787) and a fluorescent strain (TG(KDR:EGFP)). Dose-response curves and EC50 values were determined by KNIME-Workflows