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. Author manuscript; available in PMC: 2007 Sep 1.
Published in final edited form as: Prog Retin Eye Res. 2006 Sep 7;25(5):490–513. doi: 10.1016/j.preteyeres.2006.07.003

Figure 5.

Figure 5

Proteomic analysis was performed to determine whether retinal proteins are oxidatively modified during glaucomatous neurodegeneration in ocular hypertensive eyes following hypertonic saline injections into episcleral veins. Protein expression was determined by two dimensional (2D)-polyacrylamide gel electrophoresis of equally loaded protein samples obtained from ocular hypertensive and control retinas (PI, isoelectric point). Protein oxidation was determined by identifying the retinal proteins containing carbonyl groups using 2D-oxyblot analysis. Results of this in vivo study revealed that the protein modification by ROS occurs to a greater extent in ocular hypertensive eyes compared with the controls. Comparison of 2D-oxyblots with Coomassie Blue-stained 2D-gels showed that approximately 60 protein spots (out of over 400 spots) obtained using retinal protein lysates from ocular hypertensive retinas exhibited protein carbonyl immunoreactivity, which reflects oxidatively modified proteins. Following normalization of spots to their protein content measured by the intensity of Coomassie Blue staining, a significant increase was detected in carbonyl immunoreactivity of individual protein spots obtained using retinal protein lysates from ocular hypertensive eyes compared with the controls. The oxidized proteins identified through mass spectrometry included glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a glycolytic enzyme; HSP72, a stress protein; and glutamine synthetase, an excitotoxicity-related protein (Modified with permission from (Tezel et al., 2005).