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Investigate the mechanisms underlying functional differences observed in glial fibrillary acid protein (GFAP) and vimentin knockout mice.Retinal tissue was harvested from 21-day-old mice lacking GFAP and vimentin (GFAP(-/-)vim(-/-); n=6), and age-matched wild-type (wt; n=6) controls. The retinal samples were evaluated by two-dimensional polyacrylamide gel electrophoresis, SYPRO Ruby fluorescent stain, and PDQuest software image analysis. Protein spots that were differentially expressed between the two groups were processed for identification by tandem mass spectrometry, prior to further ongoing investigations with western blotting and immunocytochemistry.Thirteen protein spots were found to be differentially expressed between the GFAP(-/-)vim(-/-) and wt mouse retinas, as determined by Mann Whitney U test with a significance level at p< 0.05.The proteins identified are involved in a wide range of biological processes that include glial cell differentiation and development, neuroprotective and neurotrophic actions, redox homeostasis, cell adhesion and migration, and cytoskeletal remodeling. This study will further our understanding of the role of GFAP and vimentin in the retina, and likely provide new insights into various conditions that involve retinal cell degeneration and glial cell activation.