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Updated: Jul 7, 2025
Mechanical Vessel Injury in Zebrafish Embryos
Published on: February 17, 2015
Analysis of vascular disruption in zebrafish embryos as an endpoint to predict developmental toxicity
Julia Nöth1, Wibke Busch2, Tamara Tal2
1Department of Bioanalytical Ecotoxicology, Helmholtz Centre for Environmental Research-UFZ, Permoserstraβe 15, 04318, Leipzig, Germany. julia.noeth@ufz.de.
Abstract:
Inhibition of angiogenesis is an important mode of action for the teratogenic effect of chemicals and drugs. There is a gap in the availability of simple, experimental screening models for the detection of angiogenesis inhibition. The zebrafish embryo represents an alternative test system which offers the complexity of developmental differentiation of an entire organism while allowing for small-scale and high-throughput screening. Here we present a novel automated imaging-based method to detect the 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. Parallel assessment of arterial and venal aorta ruled out a potential bias by impaired heart or blood cell development. In contrast, the histone deacetylase inhibitor valproic acid did not affect ISV formation supporting the specificity of the angiogenic effects. 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 h post-fertilization). One of the test compounds was showing particularly high specificity for angiogenesis effects (SU4312) and was, therefore, suggested as a model compound for the identification of molecular markers of angiogenic disruption. 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.
Insights
We developed a new, automated video analysis method to detect chemical-induced inhibition of blood vessel formation (angiogenesis) in zebrafish embryos. This high-throughput screening tool is more sensitive than existing methods for identifying teratogenic compounds.
Area of Science:
- Developmental toxicology
- Vascular biology
- High-throughput screening
Background:
- Angiogenesis inhibition is a key mechanism for chemical teratogenicity.
- Existing screening models for angiogenesis inhibition lack simplicity and high-throughput capacity.
- Zebrafish embryos offer a complex yet scalable model for developmental toxicity testing.
Purpose of the Study:
- To develop and validate a novel, automated imaging-based method for detecting angiogenesis inhibition in zebrafish embryos.
- To assess the sensitivity and specificity of this new method compared to existing approaches.
- To establish a high-throughput screening system for identifying chemical disruption of angiogenesis.
Main Methods:
- Automated video subtraction analysis of blood cell movement to quantify functional intersegmental vessels (ISVs) in zebrafish embryos.
- Exposure of zebrafish embryos to various tyrosine kinase inhibitors (e.g., SU4312, SU5416, Sorafenib, PTK787) and a histone deacetylase inhibitor (valproic acid).
- Parallel assessment of arterial and venal aorta to exclude biases from cardiac or hematopoietic defects.
Main Results:
- The method successfully detected concentration-dependent inhibition of angiogenesis by tyrosine kinase inhibitors.
- Higher sensitivity (lower effect concentrations) was observed compared to a Tg(KDR:EGFP) reporter gene strain.
- Specificity for angiogenesis was enhanced when exposure began at later embryonic stages (24 hpf).
- Valproic acid did not affect ISV formation, supporting the specificity of the observed angiogenic effects.
Conclusions:
- Automated video imaging in wild-type zebrafish embryos provides a viable, non-invasive, high-throughput method for detecting chemical-induced angiogenic disruption.
- This method offers superior sensitivity and specificity for identifying teratogenic compounds affecting blood vessel development.
- SU4312 was identified as a model compound for further research into molecular markers of angiogenic disruption.
