Application of Fluorescence Difference Gel Electrophoresis Technology in Searching for Protein Biomarkers in Chick Myopia

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• 作者：Thomas C. Lam ; King-Kit Li ; Samuel C. L. Lo ; Jeremy A. Guggenheim ; Chi Ho To
• 刊名：Journal of Proteome Research
• 出版年：2007
• 出版时间：November 2007
• 年：2007
• 卷：6
• 期：11
• 页码：4135 - 4149
• 全文大小：1060K
• 年卷期：v.6,no.11(November 2007)
• ISSN：1535-3907

文摘

The lens-induced myopia (LIM) in response to concave lens (negative lens) is a well established animalmodel for studying myopia development. However, the exact visual and neurochemical signalingmechanisms involving myopic eye growth are yet to be elucidated. The feasibility of applying a noveltwo-dimensional fluorescence difference gel electrophoresis technique for global protein profilings anda search for differential protein expressions in LIM were explored in the present study. Two-dimensionalpolyacrylamide gel electrophoresis was performed employing a "minimal Lysine labeling" approachand a reverse CyeDye experimental protocol using retinal tissue from chicks. The retinal protein profilesbetween myopic and control eyes were found to be very similar. More than a thousand protein spotscould be detected on a 2D gel. Sixteen and ten protein spots were found to be up-regulated and down-regulated respectively in the myopic eyes according to our preset criteria with the inclusion of an internalpool standard. About 65% of those filtered spots could be successfully identified by peptide massfingerprinting by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry . Mostof the differentially expressed proteins were found to be related to cytoskeletal or oxidative functions.According to the prediction of subcellular locations, most of them (about 84%) were classified ascytoplasmic proteins. The cellular functions for those differentially expressed proteins were reportedand their possible involvements in the compensated eye growth were discussed. We have optimizeda workable protocol for the study of the differential retinal protein expressions in the LIM using 2D-DIGE approach which was shown to have a number of advantages over the traditional 2D electrophoresistechnique.