Comparative Proteome Analysis of Neural Retinas from Type 2 Diabetic Rats by Two-dimensional Electrophoresis

Abstract

PURPOSE: The purpose of this study was to isolate, identify, and analyze diabetes-related protein changes that occur in neural retinas in vivo. METHODS: Total proteins were extracted from neural retinas of normal and 8-weeks diabetic Sprague-Dawley (SD) rats and separated by two-dimensional gel electrophoresis (2-DE). Some protein spots exhibiting statistically significant variations (p < 0.05) were selected randomly and identified by mass spectrometry (MS or MS/MS). The protein alphaA-crystallin was chosen as a target for specific immunodetection using Western blot to corroborate the variation found by 2-DE. RESULTS: Twenty protein spots identified included alphaA-crystallin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), glutamine (Gln) synthetase, and so forth. Western blotting analyses confirmed that alphaA-crystallin protein expression was upregulated in diabetic retina. CONCLUSIONS: In this study, we isolate, identify, and analyze diabetes-related protein changes that occur in neural retinas in vivo. Further investigation of candidate proteins may identify novel pharmacological targets for diabetic retinopathy.