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Z.-L. Chi, H. Okamoto, M. Suzuki, K. Terao, Y. Yoshikawa, T. Iwata; Characterization of Retinal Pigment Epithelium Cells Isolated From Cynomolgus Monkey With Early-Onset Macular Degeneration. Invest. Ophthalmol. Vis. Sci. 2008;49(13):215.
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We have previously reported a cynomolgus (Macaca fascicularis) pedigree with early-onset macular degeneration. These monkeys show cardinal features of age-related macular degeneration in human such as pigmentary disorders and/or drusen-like spots at two years after birth (Umeda et al., 2005) and ERG abnormality. Here we report a characterization of retinal pigment epithelium (RPE) cells isolated from two affected monkeys.
RPE cells were isolated from wild type (11 and 14 yrs old) and affected monkeys with severe (17 yrs old) and modest (19 yrs old) disease progression based on fundus photograph, ERG, and fluorescence fundus photograph. The RPE cells were used for cell marker staining, proliferation assay, phagocytosis analysis (Olympus, LCV100), gene expression analysis using cDNA microarray, proteome analysis by 2D-gel electrophoresis and LC-MS/MS mass spectrometer (ThermoElectron, LCQ DECA XP Plus).
RPE cells from affected monkeys were significantly reduced of proliferation speed and phagocytosis activity. These RPE cells were also unable to form tight junction observed by Zonula Occludens-1 (ZO-1) immunostaining. Genes and proteins expression of RPE cells were significantly exhibiting significant expression changed at different disease stages compared with normal RPE cells.
In this study, primary RPE cells from affected and control monkeys were analyzed. Significant change of gene and protein expression leading to loss of tight junction in RPE cells suggests that barrier function between neural retina and choroid maybe the cause of the disease.
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