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coordinated endothelial cell activities,

The zebrafish (Danio rerio) is a small tropical freshwater fish, which is used as a model system in both drug discovery and developmental biology studies and is increasingly being used in studies on angiogenesis. The development of blood vessels such as the dorsal aorta, posterior cardinal vein, subintestinal veins and intersegmental vessels in early zebrafish embryos is well characterized through analysis of mutations affecting their formation. For example, mutations such as floating head where embryos lack the dorsal aorta and intersegmental vessels, or sonic-you where the dorsal aorta fails to undergo tubulogenesis and intersegmental vessels are absent These vessels are easily monitored , thus making them suitable for identification of angiogenesis inhibitors. Several studies have used early zebrafish embryos to show that tyrosine kinase inhibitors can prevent angiogenesis measured using either microangiography or endogenous alkaline phosphatase staining of blood vessels Transgenic zebrafish lines with fluorescent (GFP-labelled) blood vessels have also been developed, which greatly simplify the imaging of vessels

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Figure 6

Angiogenesis in the zebrafish embryo. Angiogram of a zebrafish larva at 7 day post fertilization (lateral view, anterior to the left, dorsal side up). The zebrafish fli 1 promoter was used to drive the expression of enhanced green fluorescent protein (EGFP) in all blood vessels; continuous in vivo observation of the vertebrate embryonic vasculature was achieved by use of time-lapse multiphoton laser microscopy. Scale bar, 500 µm (Image courtesy of Dr BM Weinstein, Unit on Vertebrate Organogegesis, Laboratory of Molecular Genetics, NICHD, Bethesda, MD).

Zebrafish are inexpensive to use and easy to maintain long-term, producing hundreds of offspring per mating every week, making large-scale screening experiments possible. Furthermore, as the embryos develop outside the mother and are transparent, the direct observation of blood vessel formation and subsequent blood vessel development are straightforward using just a low-power binocular microscope