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

Chuen Lam, King Kit Li, Chun Lap Samuel Lo, Jeremy A. Guggenheim, Chi Ho To

Research output: Journal article publicationJournal articleAcademic researchpeer-review

20 Citations (Scopus)

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 down-regulated 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 2D-DIGE approach which was shown to have a number of advantages over the traditional 2D electrophoresis technique.
Original languageEnglish
Pages (from-to)4135-4149
Number of pages15
JournalJournal of Proteome Research
Volume6
Issue number11
DOIs
Publication statusPublished - 1 Nov 2007

Keywords

  • Chick
  • DIGE
  • Mass spectrometry
  • Myopia
  • Proteomics
  • Retina
  • Two-dimensional gel electrophoresis

ASJC Scopus subject areas

  • Chemistry(all)
  • Biochemistry

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