Open AccessThe embryonic mouse hindbrain as a qualitative and quantitative model for studying the molecular and cellular mechanisms of angiogenesis
Author(s) -
Alessandro Fantin,
Joaquim Miguel Vieira,
Alice Plein,
Charlotte H. Maden,
Christiana Ruhrberg
Publication year - 2013
Publication title -
nature protocols
Language(s) - English
Resource type - Journals
eISSN - 1754-2189
pISSN - 1750-2799
DOI - 10.1038/nprot.2013.015
Subject(s) - hindbrain , angiogenesis , immunolabeling , biology , embryonic stem cell , microbiology and biotechnology , vasculogenesis , embryo , blood vessel , zebrafish , multicellular organism , sprouting angiogenesis , neovascularization , anatomy , neuroscience , genetics , immunology , immunohistochemistry , gene , progenitor cell , stem cell , endocrinology
The mouse embryo hindbrain is a robust and adaptable model for studying sprouting angiogenesis. It permits the spatiotemporal analysis of organ vascularization in normal mice and in mouse strains with genetic mutations that result in late embryonic or perinatal lethality. Unlike postnatal models such as retinal angiogenesis or Matrigel implants, there is no requirement for the breeding of conditional knockout mice. The unique architecture of the hindbrain vasculature allows whole-mount immunolabeling of blood vessels and high-resolution imaging, as well as easy quantification of angiogenic sprouting, network density and vessel caliber. The hindbrain model also permits the visualization of ligand binding to blood vessels in situ and the analysis of blood vessel growth within a natural multicellular microenvironment in which endothelial cells (ECs) interact with non-ECs to refine the 3D organ architecture. The entire procedure, from embryo isolation to imaging and through to results analysis, takes approximately 4 d.