Quantitative assessment of angiogenesis, perfused blood vessels and endothelial tip cells in the postnatal mouse brain

Item Type:	Article
Title:	Quantitative assessment of angiogenesis, perfused blood vessels and endothelial tip cells in the postnatal mouse brain
Creators Name:	Wälchli, T. and Mateos, J.M. and Weinman, O. and Babic, D. and Regli, L. and Hoerstrup, S.P. and Gerhardt, H. and Schwab, M.E. and Vogel, J.
Abstract:	During development and in various diseases of the CNS, new blood vessel formation starts with endothelial tip cell selection and vascular sprout migration, followed by the establishment of functional, perfused blood vessels. Here we describe a method that allows the assessment of these distinct angiogenic steps together with antibody-based protein detection in the postnatal mouse brain. Intravascular and perivascular markers such as Evans blue (EB), isolectin B4 (IB4) or laminin (LN) are used alongside simultaneous immunofluorescence on the same sections. By using confocal laser-scanning microscopy and stereological methods for analysis, detailed quantification of the 3D postnatal brain vasculature for perfused and nonperfused vessels (e.g., vascular volume fraction, vessel length and number, number of branch points and perfusion status of the newly formed vessels) and characterization of sprouting activity (e.g., endothelial tip cell density, filopodia number) can be obtained. The entire protocol, from mouse perfusion to vessel analysis, takes ∼10 d.
Keywords:	Angiogenesis, Immunohistochemistry, Animals, Mice
Source:	Nature Protocols
ISSN:	1754-2189
Publisher:	Nature Publishing Group
Volume:	10
Number:	1
Page Range:	53-74
Date:	January 2015
Official Publication:	https://doi.org/10.1038/nprot.2015.002
PubMed:	View item in PubMed

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