Abstract
PURPOSE: Given the critical role of the retinal pigment epithelium (RPE) in the pathogenesis of age-related macular generation (AMD) and the links drawn between chaperone proteins and neurodegenerative disease, we aimed to culture RPE from the Ccl2(-/-)/Cx3cr1(-/-) mouse model of AMD and evaluate expression of chaperone protein ERp29. MATERIALS AND METHODS: Murine RPE cells were surgically and chemically isolated and cultured. ERp29 mRNA and protein expression were evaluated by real-time RT-PCR, immunohistochemistry, and Western blot. RESULTS: Ccl2(-/-)/Cx3cr1(-/-) RPE was successfully isolated and cultured. They presented a decreased but statistically insignificant difference in ERp29 transcript and protein expression compared to C57B6/L wild type mouse RPE. CONCLUSIONS: The effective murine RPE culture described here enables future investigation into RPE biology and AMD pathogenesis. Although we found a decrease in ERp29 expression in the Ccl2(-/-)/Cx3cr1(-/-) RPE, the difference was less than we previously observed in the whole retina. This suggests that the RPE may not contribute to the greater differential expression and ERp29 might have a more significant role in the neuroretina than in the RPE during AMD pathogenesis.