Computer-Assisted Semi-Quantitative Analysis of Mouse Choroidal Density
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.
KeywordsVascular Endothelial Growth Factor White Area Geographic Atrophy Choroidal Vessel Choroidal Blood Flow
I thank W. Zheng, Dr. L. Zheng, and M. Zhu for technical assistance and Dr. M. Tanito for the tutorial of Adobe Photoshop program. This study was supported by NIH grants RR17703, and EY12190, ADA grant 1-06-RA-76, AHAF grant M2008-059, FFB grant BR-CMM-0808-0453-UOK and unrestricted grants from Hope for Vision and Research to Prevent Blindness.