Angiogenesis Protocols

Emma Ristori, Sandra Donnini, Marina Ziche

The zebrafish is an excellent animal model to study the formation of the vertebrate vascular network. The small size, the optical translucency, and the ability to model endothelial-specific fluorescent transgenic lines in the zebrafish embryo had facilitate, in the past 10 years, the direct visualization of vessels formation and remodeling. Furthermore, zebrafish is an excellent disease model such as for cancer and neurodegenerative diseases. Cerebral amyloid angiopathy (CAA) is a human neurovascular degenerative disease, caused by Amyloid β (Aβ) peptides deposition around brain microvessels, and characterized by vascular brain degenerative changes. By using the zebrafish model, we investigated the effect of Aβ peptides treatment in vessel formation during embryogenesis. We showed that the defects in the vascular remodeling and senescence can be detected, respectively, via staining for alkaline phosphatase activity and β-galactosidase or cyclin-dependent kinase inhibitor p21 expression. We demonstrated that treating zebrafish embryos with these oxidative peptides reduces angiogenesis and promotes premature vascular senescence. In this chapter, we will describe the methods to reveal both angiogenesis and senescence defects upon Aβ peptides treatment of the zebrafish embryos.