Two-dimensional electrophoregram for proteomic analysis of rat brain with intrahippocampal amyloid β injection and normal rat brain

Abstract

Abstract: Objective To investigate the molecular mechanisms of Alzheimer’s disease by comparing global protein patterns in two-dimensional electrophoregram (2-DE) of the brain of rats with intrahippocampal amyloid β injection and normal rats. Methods From adult SD rats with intrahippocampal injection of amyloid β, 200 μg brain proteins were extracted with 9 mol/L urea, 4% CHAPS, 1% DTT, 0.5% CA and a cocktail of protease inhibitors. Immobilized pH gradient (IPG) isoelectric focusing electrophoresis of the extracted proteins was performed to obtain the electrophoretogram of the first dimension, with the second dimension obtained by vertical SDS-PAGE. The electrophoretograms were visualized using silver staining and analyzed with ImageMaster 2D-Elite software. Results On average, 496 and 491 protein spots could be obtained in the electrophoregraphs for rats with amyloid β and the control rats, respectively, and 30 of these spots exhibited quantitative changes. Another 11 and 6 spots were exclusively shown on the protein maps for amyloid β-treated rats and control rats, respectively. Conclusion The differentially displayed proteins in the brain identified between the rats with intrahippocampal amyloid β injection and control rats may provide further insight into the pathogenesis of Alzheimer’s disease and useful clues for developing new drugs for its treatment.

CLC Number:

R742.89
R592

YANG Guo-feng¹, WANG Lu-ning¹, ZHAO Xin², NIE Yong-hui¹. Two-dimensional electrophoregram for proteomic analysis of rat brain with intrahippocampal amyloid β injection and normal rat brain[J]. Journal of Southern Medical University, 2004, 24(05): 553-555.

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