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C. Lam, K.K. Li, S.C. L. Lo, C.H. To; Differential Protein Expressions in the Emmetropization of Chick Retina by a Proteomic Approach . Invest. Ophthalmol. Vis. Sci. 2005;46(13):3144.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To examine and identify the retinal protein expressions in various developmental stages of the white Leghorn chick. To investigate the differential retinal protein expressions in response to the lens–induced defocus. Methods: The retinal tissues were harvested from normal developing chicks (3 day–, 10 day– and 20 day–old) and also from emmetropizing retinal tissues under +10D & –10D lens treatments. Protein profiles were generated by two–dimensional gel electrophoresis (2DE) and visualized by either Coomassie Blue stain or Mass Spectrometry (MS) compatible silver stain. The 2DE retinal protein maps generated were compared and analyzed using ImageMasterTM 5 gel matching software. Protein identification for Coomassie stained gels were processed and done in an automatic robotic platform. A number of the differentially expressed spots detected by silver stain were excised manually and trypsin digested. Mass spectra were acquired from a Bruker Autoflex MALDI–TOF MS and the resulting peptide maps were searched against the NCBInr databases via the Mascot search engine. Results: Over 150 soluble chick retinal proteins within pH3–10 were currently identified and were classified according to their molecular functions. The major proteins were found involved in catalytic activity (∼40%) and binding (∼30%). 2D gels profile analysis showed a strong similarity in the protein profiles among the three groups of developing chick retinae. 3 protein spots were found to be down–regulated while another 4 protein spots were up–regulated over the time period. Besides, one extra protein could only be resolved and detected in using narrower pH5–8 IPG but not in the pH3–10 range. Among them, 2 down–regulated proteins and 3 up–regulated proteins could be successfully identified by MALDI–TOF peptide mass fingerprinting. For retinal samples under the treatment of optic defocus, two candidate differentially expressed protein spots which related to growth were detected. Conclusions: In the present study, we have shown the differential protein expressions in 3 day–, 10 day– and 20 day–old chick retinae using the 2DE technique. Some differentially expressed proteins involved in normal development and induced compensation growth were successfully identified with MALDI–TOF MS.
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