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F. Altmann, S.M. Hauck, S. Schoeffmann, H. Gerhards, B. Kaspers, M. Ueffing, C.A. Deeg; Comparison of the Vitreous Proteome of ERU Diseased and Visually Healthy Horse Eyes . Invest. Ophthalmol. Vis. Sci. 2006;47(13):4528.
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Equine recurrent uveitis (ERU), a spontaneous disease with high incidence, is a valuable model for recurrent uveitis. To interrupt the cycle of repeated recurrences, the vitreous can be removed surgically by pars–plana vitrectomy. In this study, we were interested in the identification of differentially expressed proteins in vitreous of horses with ERU in comparison to vitreous of visually healthy horses, in order to detect pathways and factors involved in the pathogenesis of recurrent uveitis.
Vitreous samples were stabilized with protease inhibitors, lyophilized and dialyzed. 2DE was done by IEF (IPG strips, pH 3–11) on a Multiphor for 14200 Vh at 20°C, and subsequent electrophoreses in gradient SDS–PAGE gels (9–15%). The gels were then silver–stained and spots were excised and prepared for mass spectrometry. We analyzed the vitreous proteome of 12 visually healthy eyes and 24 eyes with recurrent uveitis. Spots of the healthy vitreous proteome and differentially expressed spots were identified using MALDI–TOF.
Vitreous of ERU diseased eyes showed an increased protein content (average protein concentration in vitreous of uveitic horses 3,70 µg/µl; range from 0,40 to 9,80 µg/µl; n=30) compared to the negative controls (average protein concentration 0,16 µg/µl; range from 0,10 to 0,42 µg/µl; n=19). Protein patterns appeared very homogenous within the group of diseased eyes (n=24) but distinguished obviously from those of controls (n=12) which also showed very similar patterns within their group. In 2DE, 6 proteins were higher abundant in diseased vitreous compared to controls. 4 proteins showed higher abundance in controls than in ERU vitreous. The differentially expressed proteins can be functionally classified as serum derived, inflammatory related, neuroprotective factors, acting in maintainance of the blood–retina barrier, associated with apoptotic pathways and playing a role in vascularization.
Proteome analysis is useful to study mechanisms involved in uveitis pathogenesis.
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