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S Roth, TK Ro, AR Shaikh, SL Tollaksen, C Zhang, CS Giometti; Changes in Protein Expression after Retinal Ischemic Preconditioning: A Proteomics Approach . Invest. Ophthalmol. Vis. Sci. 2002;43(13):784.
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Purpose: Ischemic preconditioning (IPC) protects the rat retina against the injury that ordinarily follows severe ischemia. We showed previously that release of adenosine, de novo protein synthesis, and mediators such as protein kinase C and K+ATP channels are required for IPC protection (1-3). This study used a proteomics approach to examine for candidate proteins that may be involved in IPC. Methods: An ischemic preconditioning stimulus was applied in anesthetized Sprague-Dawley rats. At various times following IPC, retinas were homogenized and proteins separated by two-dimensional gel electrophoresis (2DE). Protein bands were cut from Coomassie Blue-stained gels, digested in situ with trypsin, and identified via Sequest® database searching of uninterpreted tandem mass spectra obtained from a Q-Tof mass spectrometer (4). Identitification and degree of change in expression of some of the proteins was confirmed using Western blotting. Results: The expression of nearly 60 proteins was altered after IPC. All 7 proteins isolated and submitted for Q-Tof were identified. Creatine kinase, enolase, G protein ß subunit 1, and GFAP increased after IPC; identities of the latter 2 were confirmed by Western blotting. Alpha and beta crystallins decreased, also confirmed by Western blotting. Conclusion: IPC alters protein synthesis. G-proteins, GFAP, and alpha crystallin are involved in this neuroprotective effect. Results show that 2DE and proteomics constitute a method for elucidating mechanisms of IPC. References: 1) Roth S, et al: IOVS 1998; 39: 775-785. 2) Li B, et al: IOVS 1999; 40: 1200-1216. 3) Li B et al: Exp Eye Res 2000; 70: 755-765. 4) Stone KL et al: Electrophoresis 1998; 19: 1046-1052.
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