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N. Mandal, G. P. Lewis, S. K. Fisher, S. Heegaard, J. U. Prause, B. Honoré, H. Vorum, M. la Cour; Retinal Proteomic Changes Following Experimental Retinal Detachment. Invest. Ophthalmol. Vis. Sci. 2009;50(13):3184.
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To determine the protein profile changes of the retina following experimental detachment using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE).
An inferior retinal detachment was induced in the right eye of six New Zealand Red pigmented rabbits by infusing a solution of sodium hyaluronate (Healon, 0.25% in balanced salt solution) via a glass pipette between the neural retina and retinal pigment epithelium (RPE). Healon was necessary to prevent spontaneous retinal reattachment. Sham surgery, surgical entry of the vitreous cavity with no disruption of the retina, was performed in the right eye of six other rabbits that were used as controls. At seven days the eyes were enucleated and the inferior detached and attached retinas from the experimental and control animals respectively were peeled from the RPE and immediately snap frozen in liquid nitrogen in separate vials. Retinal samples were subjected to high-resolution 2D-PAGE. Following visualisation of the protein spots by silver staining, gels were scanned and the pattern of protein expression analysed with the PDQuest software program. Spots of interest were excised from the gels and processed by tandem mass spectrometry.
Approximately 800 protein spots were clearly resolved by 2D-PAGE. The overall protein expression profiles of the twelve retinas were similar. Eighteen protein spots were found to be at least two-fold differentially expressed (p<0.05 Mann-Whitney U test) between the detached and attached retinas. Nine protein spots were downregulated and nine upregulated in the detached retinal group. The proteins identified had a broad spectrum of function, which included signal transduction, energy production and structure.
The differentially expressed proteins elucidated in this study may play an important role in the cellular responses of the retina to detachment, its subsequent ability to recover after reattachment as well as visually devastating complications such as proliferative vitreoretinopathy or subretinal fibrosis.
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