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Evaluating Vascular Hyperpermeability-inducing Agents in the Skin with the Miles Assay

Brash, JT; Ruhrberg, C; Fantin, A; (2018) Evaluating Vascular Hyperpermeability-inducing Agents in the Skin with the Miles Assay. Journal of Visualized Experiments (136) , Article e57524. 10.3791/57524. Green open access

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Abstract

The primary function of the vascular endothelium in vertebrate organisms is to serve as a barrier between the blood and each tissue of the body, whereby the permeability of the endothelium to blood cells, plasma macromolecules, and water can be adapted according to the physiological need. In certain diseases, cytokines and growth factors are released that target the endothelial barrier to transiently increase vascular permeability; however, their prolonged presence may cause chronic vascular hyperpermeability and thereby tissue-damaging edema. The Miles assay is an in vivo technique that allows researchers to study vascular hyperpermeability through the proxy measurement of vascular leakage. Here, we provide a detailed protocol on how to perform this procedure in the mouse, which is the most widely used model organism to study mammalian physiology and pathology. The procedure involves the intravenous injection of Evans blue dye to label the circulating albumin followed by multiple intradermal injections of permeability-inducing agents and vehicle control solutions into opposing flanks of the mouse. Consequently, Evans blue dye gradually leaks into the dermis, where it accumulates and can be extracted for quantification as leakage induced by the permeability-inducing agent relative to the vehicle. The Miles assay can be performed in wild type or genetically modified mouse models and may be combined with drug administration to study molecular mechanisms that regulate vascular permeability and identify agents/targets capable of inducing or blocking hyperpermeability.

Type: Article
Title: Evaluating Vascular Hyperpermeability-inducing Agents in the Skin with the Miles Assay
Open access status: An open access version is available from UCL Discovery
DOI: 10.3791/57524
Publisher version: http://dx.doi.org/10.3791/57524
Language: English
Additional information: This article is published under a Creative Commons licence CC-BY-NC-ND 3.0 (https://creativecommons.org/licenses/by-nc-nd/3.0/legalcode)
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > Institute of Ophthalmology
URI: https://discovery.ucl.ac.uk/id/eprint/10050926
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