A Proteomic Approach to PVR: Comparative Proteome Analysis of Native, Differentiated and Cultured, Dedifferentiated Human RPE Cells
Alge C. S.1,2, Suppmann S. 2,3, Priglinger S. G.1, Neubauer A. S.1, Welge-Lussen U.1, Ueffing M. 2,3, Kampik A. 1,2
1Department of Ophthalmology, Ludwig-Maximilians University, Munich; 2Clinical Cooperation Group Ophthalmogenetics, Ludwig-Maximilians University; 3GSF-Research Center for Environment and Health, Oberschleißheim
Purpose: Dedifferentiation of retinal pigment epithelial (RPE) cells is considered to be a crucial event in the pathogenesis of proliferative vitreoretinopathy (PVR). In an attempt to better understand the pathophysiology of early PVR, the protein expression pattern of native, differentiated RPE cells was to that of cultured, thereby dedifferentiated RPE cells.
Method: Differentiated, native human RPE cells and dedifferentiated, cultured human RPE cells were analyzed by 2-D electrophoresis. Total cellular proteins were separated by isoelectric focusing using immobilized pH gradients (IPG 3-10) and electrophoresis on 9 - 15% gradient polyacrylamide gels. Proteins were visualized by silver staining. 2-DE gels of both cell-types were then compared and spots of interest were analyzed by MALDI-TOF (matrix-assisted laser desorption ionization time of flight) mass spectroscopy. The resulting peptide mass fingerprints were searched against the NCBInr, MSDB and EnsemblC database to identify the respective prote
Zurück | Back