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Application of fluorescence difference gel electrophoresis technology in searching for protein biomarkers in chick myopia

Lam, Thomas C., Li, King-Kit, Lo, Samuel C. L., Guggenheim, Jeremy Andrew and To, Chi Ho 2007. Application of fluorescence difference gel electrophoresis technology in searching for protein biomarkers in chick myopia. Journal of Proteome Research 6 (11) , pp. 4135-4149. 10.1021/pr0701097

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Abstract

The lens-induced myopia (LIM) in response to concave lens (negative lens) is a well established animal model for studying myopia development. However, the exact visual and neurochemical signaling mechanisms involving myopic eye growth are yet to be elucidated. The feasibility of applying a novel two-dimensional fluorescence difference gel electrophoresis technique for global protein profilings and a search for differential protein expressions in LIM were explored in the present study. Two-dimensional polyacrylamide gel electrophoresis was performed employing a “minimal Lysine labeling” approach and a reverse CyeDye experimental protocol using retinal tissue from chicks. The retinal protein profiles between myopic and control eyes were found to be very similar. More than a thousand protein spots could be detected on a 2D gel. Sixteen and ten protein spots were found to be up-regulated and downregulated respectively in the myopic eyes according to our preset criteria with the inclusion of an internal pool standard. About 65% of those filtered spots could be successfully identified by peptide mass fingerprinting by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry . Most of 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 as cytoplasmic proteins. The cellular functions for those differentially expressed proteins were reported and their possible involvements in the compensated eye growth were discussed. We have optimized a workable protocol for the study of the differential retinal protein expressions in the LIM using 2DDIGE approach which was shown to have a number of advantages over the traditional 2D electrophoresis technique.

Item Type: Article
Status: Published
Schools: Optometry and Vision Sciences
Uncontrolled Keywords: Two-dimensional gel electrophoresis ; chick ; myopia ; proteomics ; DIGE ; retina ; mass spectrometry
Publisher: American Chemical Society
ISSN: 1535-3893
Last Modified: 04 Jun 2017 01:58
URI: http://orca.cf.ac.uk/id/eprint/6878

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