Fibroblast growth factor 2-induced angiogenesis in zebrafish: the zebrafish yolk membrane (ZFYM) angiogenesis assay
Open accessAbstract
Angiogenesis plays a key role in tumour 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 hrs after fertilization. The 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.
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Human FGF2 activates the zebrafish Fgf2 receptor.
“Human and zebrafish FGF2 exhibit ~75% identity and human FGF2 activates the zebrafish Fgf2 receptor (Nicoli et al., 2009).”
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“Human and zebrafish FGF2 exhibit ~75% identity and human FGF2 activates the zebrafish Fgf2 receptor (Nicoli et al., 2009).”
BackgroundVEGF-B prevents excessive angiogenesis by inhibiting FGF2/FGFR1 pathway
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- Authors
- Nicoli, S., De Sena, G., & Presta, M.
- Affiliations
- University of Massachusetts Chan Medical School (US)
- University of Brescia (IT)
- Corresponding author
- Presta, Marco
- Venue
- Journal of Cellular and Molecular Medicine
- Publisher
- Wiley
- Published
- 2008-07-24
- Volume
- 13
- Issue
- 8b
- Pages
- 2061-2068
- ISSN
- 1582-1838 (ISSN-L)
- 1582-4934
- PMC
- PMC6512384
- OpenAlex
- W1997053062
- Cited by
- 74
- References
- 37
- Citations by year
- 2026: 1 · 2023: 1 · 2022: 12 · 2021: 7 · 2020: 4 · 2019: 1 · 2018: 2 · 2017: 5 · 2016: 2 · 2015: 6 · 2014: 7 · 2013: 4 · 2012: 6
- FWCI
- 2.19
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Journal of Cellular and Molecular Medicine (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Institutional Research Information System (Università degli Studi di Brescia) (submittedVersion)
- Funders
- Istituto Superiore di Sanità
- Associazione Italiana per la Ricerca sul Cancro
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Zebrafish Biomedical Research Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Angiogenesis and VEGF in Cancer
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fibroblast Growth Factor Research
- Keywords
- angiogenesis, fibroblast growth factor 2, FGF receptor 1, SU5402, pentraxin 3, VEGF-A, SU5416, endothelial cell proliferation, microangiography, zebrafish embryos
- MeSH terms
- Animals, Yolk Sac, Zebrafish, Fibroblast Growth Factor 2, Neovascularization, Physiologic
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