PUBLICATION
Fibroblast Growth Factor 2-induced angiogenesis in zebrafish: the zebrafish yolk membrane (ZFYM) angiogenesis assay
- Authors
- Nicoli, S., De Sena, G., and Presta, M.
- ID
- ZDB-PUB-080801-3
- Date
- 2009
- Source
- Journal of Cellular and Molecular Medicine 13(8b): 2061-2068 (Journal)
- Registered Authors
- Nicoli, Stefania, Presta, Marco
- Keywords
- zebrafish, angiogenesis, FGF2, inhibitors, embryo
- MeSH Terms
-
- Animals
- Fibroblast Growth Factor 2/physiology*
- Neovascularization, Physiologic/physiology*
- Yolk Sac/blood supply*
- Zebrafish
- PubMed
- 18657228 Full text @ J. Cell. Mol. Med.
Citation
Nicoli, S., De Sena, G., and Presta, M. (2009) Fibroblast Growth Factor 2-induced angiogenesis in zebrafish: the zebrafish yolk membrane (ZFYM) angiogenesis assay. Journal of Cellular and Molecular Medicine. 13(8b):2061-2068.
Abstract
Angiogenesis plays a key role in tumor growth and metastasis. The teleost zebrafish (Danio rerio) represents a promising alternative model in cancer research. Here, we describe a zebrafish yolk membrane (ZFYM) angiogenesis assays based on the injection of 1-30 ng of human recombinant FGF2 (rFGF2) in the perivitelline space of zebrafish embryos in the proximity of developing subintestinal vein vessels (SIVs) at 48 hours post-fertilization. rFGF2 induces a rapid and dose-dependent angiogenic response from the SIV basket, characterized by the ectopic growth of newly formed, alkaline phosphatase-positive blood vessels. These vessels are formed by proliferating cells that incorporate bromodeoxyuridine and express the endothelial cell markers vegfr2/kdr and fli1. Microangiography shows that rFGF2-induced vessels are patent and connected to the systemic circulation of the embryo. In keeping with these observations, fli1:EGFP(+) cells isolated from transgenic tg(fli1:EGFP)(y1) zebrafish embryos express the tyrosine-kinase (TK) FGF receptor-1 (FGFR1) and activate extracellular signal-regulated kinase signalling when stimulated in vitro by rFGF2. The low molecular weight TK-FGFR1 inhibitor SU5402 and the high molecular weight FGF2 antagonist long-pentraxin 3 inhibit the angiogenic activity of rFGF2 when added to fish water or when co-injected with the growth factor, respectively. Moreover, similar to rFGF2, injection of the zebrafish form of vascular endothelial growth factor-A (VEGF-A) induces a significant angiogenic response in the ZFYM assay that is suppressed by the VEGF receptor-2/KDR TK inhibitor SU5416. The ZFYM assay represents a novel tool for testing the activity of low and high molecular weight inhibitors targeting a defined angiogenic growth factor in zebrafish. The assay may offer significant advantages when compared to other animal models.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping