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S Suppmann, H Zischka, J Schoch, S Hauck, T Meitinger, M Ueffing; A High Resolution Protein Map of Human Neural Retina in Comparison to Murine, Bovine and Avian Retinae . Invest. Ophthalmol. Vis. Sci. 2002;43(13):3632.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To study the protein expression pattern of human neural retina and compare its protein profile to that of the mouse, bovine and chicken. Methods: Retinal tissues from a human donor eye, C57 BL/6 mice, bovine and chicken were dissected and frozen immediately, or processed directly after dissection. Tissue preparation was optimized to maximize protein yield and to minimize proteolytic degradation. Lyzed retinae were subjected to broad range IEF (IPG 3-10) and high resolution IEF (IPG 3-6, 5-8, 7-10), and run on 9%-16% gradient SDS-PAGE in order to visualize proteins ranging from 14 to 200 kD. Gels were silver stained and analyzed using the Z3 Image Analysis software. Spots were excised and subjected to matrix-assisted laser desorption ionisation time of flight mass spectroscopy (MALDI-TOF). The Mascot software was used to search tryptic peptide fingerprints in MSDB database and in a retina-specific cDNA database, mined from UniGene with a specialized in house software. Results: To date we have analyzed 500 protein spots from the four mammalian species by means of peptide mass fingerprinting. The power of this technology for use in animal models with limited database sequence information, such as for bovine and chicken, will be discussed. A catalogue of proteins classified into retina-specific function - eg arrestin, IRBP, recoverin -, metabolic, signal transduction, antioxidative, will be presented, including proteins thus far not known to be expressed in retina. Both differences and matches regarding expression levels and chemical properties, especially between rod and cone-rich retinae, will be discussed. The retinal protein profile method will be compared to already published retina specific cDNA expression profiles, to determine the extent to which cDNA levels can reflect protein expression levels. Conclusion: A detailed physical map of the retinal proteome will be presented, including a catalogue of highly expressed proteins together with a comprehensive comparison of human versus mouse, bovine and chicken retina.
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