Constancy of ERp29 Expression in Cultured Retinal Pigment Epithelial Cells in the Ccl2/Cx3cr1 Deficient Mouse Model of Age-Related Macular Degeneration

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.

KEYWORDS:

• age-related macular degeneration
• animal model
• cell culture
• ERp29
• retinal pigment epithelium