Cited by 74Zebrafish Biomedical Research ApplicationsAngiogenesis and VEGF in CancerFibroblast Growth Factor Research

Fibroblast growth factor 2-induced angiogenesis in zebrafish: the zebrafish yolk membrane (ZFYM) angiogenesis assay

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Nicoli, Stefania, De Sena, Giulia, Presta, Marco·2009·Journal of Cellular and Molecular Medicine

Abstract

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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Crossref · 10.1111/j.1582-4934.2008.00432.x

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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
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
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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