Computer-Assisted Semi-Quantitative Analysis of Mouse Choroidal Density

Abstract

Geographic atrophy is a dry form of age-related macular degeneration (AMD) and a leading cause of blindness in the United States. The mechanism of the disease is unknown and there is no treatment for the disease at present. During aging and the development of geographic atrophy, there is a significant decrease in choroidal density. Since mouse is the only mammal that allows precise genomic manipulation, in vivo studies with genetically altered mice are likely to provide more mechanistic insights about the pathogenic mechanisms of the disease. To establish an efficient and quantitative procedure measuring choroidal density in mice for studies related to choroidal biology and geographic atrophy, we developed a computer-assisted semi-quantitative procedure for mouse choroidal density. In this study, mouse choroidal vessels were immunostained with anti-CD31 antibody and were detected by fluorescently labeled secondary antibody. Confocal or fluorescent microscopic images were analyzed with Adobe Photoshop software to determine the relative density of choroidal vessels. This procedure is relatively simple to perform and can be utilized to measure choroidal density efficiently in mouse models, which may be useful for preclinical studies relevant to the pathogenic mechanisms and therapeutics of geographic atrophy.