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R. Chun, K. Li, T. Lam, C. To; Apolipoprotein A1 Expression in Form Deprived Chick Retina. Invest. Ophthalmol. Vis. Sci. 2007;48(13):4169.
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To study the expression of apolipoprotein A-1 in form deprived chick retina using proteomic approach and to understand the biochemical mechanisms of eye growth in myopia.
Occluders were mounted via Velcro ring in front of right eye of 2-day-old chicks for 3 days. After 3 days of treatment, the refractive errors of the eyes were determined by retinoscopy. Retinae were harvested and the retinal proteins were extracted with detergent cocktail. The retinal proteins were separated by 12% continuous gels at pH 5-8 using fluorescent 2-dimensional differential in-gel electrophoresis (2D-DIGE). Fluorescent CyDye (Cy2) was used to label the internal standard while myopic and control retinal proteins were labeled by Cy3 or Cy5 randomly. Three gel images were captured by different excitation wavelengths in the Typhoon Scanner. Biological variations of the protein expression level were analyzed with DeCyder software. Protein spots were then visualized with sliver staining. Those differentially expressed protein spots were excised from the gel and digested with trypsin. Peptide masses of the protein digest were identified by Matrix Assisted Laser Desorption Ionization-Time of Flight (MALDI-TOF) mass spectrometry.
After analysis by DeCyder software, differentially expressed retinal proteins were identified (at least 1.2 fold difference) by MALDI-TOF mass spectrometry. The profile of differentially expressed proteins between form deprived and control retinae revealed apolipoprotein A-1 as down-regulated.
It has been found earlier that apolipoprotein A-1 was differentially expressed in lens induced myopic retina. Similar expression pattern was found in form deprived retina in the present study. It suggested that irrespective of methods of myopia induction in chicks, the accelerated ocular growth process appeared to share similar biochemical pathways that involve apolipoprotein A-1.
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