Protein Analysis of Human RPE Cells Affected by Aging and Age-Related Macular Degeneration

Abstract

In the United States, Age-Related Macular Degeneration is the leading cause of blindness in people over the age of 65 (Friedman et al., 2004). A key to understanding AMD is examining the Retinal Pigment Epithelium (RPE), a single layer of cells that form a portion of the blood/retina barrier. Previous work in our lab has shown that there are significant changes in protein content and increased mitochondrial DNA damage in RPE cells affected by AMD, suggesting that mitochondrial dysfunction plays a role in the pathology of the disease. Our hypothesis is that RPE cells from AMD donor eye samples will show protein expression consistent with mitochondrial dysfunction. In a previous analysis of human RPE cells, proteins were extracted and separated using 2-D gels. The spot intensities were analyzed and showed changes in protein content that were consistent with the hypothesis. The differences in protein content were also used to compare normal to pathologic aging. The purpose of this study was to confirm these results for select proteins using 1-D Western Blotting.