Three-dimensional imaging of intraplaque neovascularization in a mouse model of advanced atherosclerosis

Multiple lines of evidence suggest that intraplaque (IP) neovascularization promotes atherosclerotic plaque growth, destabilization and rupture. However, pharmacological inhibition of IP neo-vascularization remains largely unexplored due to the limited number of animal models that develop IP neovessels and the lack of reliable methods for visualizing IP angiogenesis. Here, we applied 3D confocal microscopy with an optimized tissue-clearing process, termed iDISCO (immunolabeling-enabled 3D Imaging of Solvent Cleared Organs) to visualize IP neovessels in ApoE-/- mice carrying a heterozygous mutation (C1039+/-) in the fibrillin-1 gene. Unlike regular ApoE-/- mice, this mouse model is characterized by the presence of advanced plaques with evident IP neovascularization. Plaques were stained with antibodies against endothelial marker CD31 for 3 days, followed by incubation with fluorescently labelled secondary antibodies. Subsequent tissue clearing with dichloromethane (DCM)/methanol, DCM and dibenzyl ether allowed easy visualization and 3D reconstruction of the IP vascular network while plaque morphology remained intact.

Publication year:2020

Keywords:A1 Journal article

BOF-keylabel:yes

BOF-publication weight:1

CSS-citation score:1

Authors:International

Authors from:Higher Education

Accessibility:Open