May 2003
Volume 44, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2003
Proteome Analysis of the Retina
Author Affiliations & Notes
  • M. Ueffing
    Institute for Human Genetics, GSF and KKG Ophthalmogenetics, Klinikum der LMU, München, Germany
  • S. Hauck
    Institute for Human Genetics, GSF and KKG Ophthalmogenetics, Klinikum der LMU, München, Germany
  • M. Swiatek
    Institute for Human Genetics, GSF and KKG Ophthalmogenetics, Klinikum der LMU, München, Germany
  • C. Alge
    University Eye Hospital, Ludwig Maximilians Universität, München, Germany
  • S. Suppmann
    University Eye Hospital, Ludwig Maximilians Universität, München, Germany
  • S. Schöffmann
    University Eye Hospital, Ludwig Maximilians Universität, München, Germany
  • D. Pompetzki
    Institute of Animal Physiology, Ludwig Maximilians Universität, München, Germany
  • C. Deeg
    Institute of Animal Physiology, Ludwig Maximilians Universität, München, Germany
  • U. Welge-Lüssen
    Institute of Animal Physiology, Ludwig Maximilians Universität, München, Germany
  • A. Kampik
    Institute of Animal Physiology, Ludwig Maximilians Universität, München, Germany
  • T. Meitinger
    Institute of Animal Physiology, Ludwig Maximilians Universität, München, Germany
  • Footnotes
    Commercial Relationships  M. Ueffing, None; S. Hauck, None; M. Swiatek, None; C. Alge, None; S. Suppmann, None; S. Schöffmann, None; D. Pompetzki, None; C. Deeg, None; U. Welge-Lüssen, None; A. Kampik, None; T. Meitinger, None.
  • Footnotes
    Support  EU grants PRORET QLK6-CT-2000-00569, PRO-AGE-RET QLK6-CT-2001-00385 and BMBF 031U108A/031U208A
Investigative Ophthalmology & Visual Science May 2003, Vol.44, 2255. doi:
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      M. Ueffing, S. Hauck, M. Swiatek, C. Alge, S. Suppmann, S. Schöffmann, D. Pompetzki, C. Deeg, U. Welge-Lüssen, A. Kampik, T. Meitinger; Proteome Analysis of the Retina . Invest. Ophthalmol. Vis. Sci. 2003;44(13):2255.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Abstract: : Purpose: Proteomic techniques allow for the visualization of complex protein expression patterns and identification of individual proteins by mass spectrometry. Methods: Four approaches to analyze aspects of retinal physiology will be presented, each based on high-resolution protein separation by two-dimensional electrophoresis, and protein identification by MALDI-TOF mass spectrometry. Results: 1: Proteome mapping: By comparative mapping of human, murine, bovine and avian (chicken) neural retinas by 2-D gel, we identified the 100 most abundant protein spots from a database generated in-house. Using a combination of biochemical and proteomic methods we have further analysed membrane proteins as well as protein-protein interaction patterns in mammalian photoreceptors. 2: Trans-differentiation in culture: Analysis of protein expression patterns from isolated primary retinal Müller glia cells in culture reveals the stability of protein expression during initial culturing, but shows loss of Müller glia cell-specific proteins, such as glutamate synthetase over time. The changes in expression reflect the trans-differentiation from a multifunctional, highly differentiated phenotype to a de-differentiated monolayer of cells in culture. 3: Secreted factors: Müller glia cells secrete factors that show a pronounced effect on photoreceptor survival by yet unknown means. A very sensitive method for searching secreted proteins from conditioned media using an in vivo 35-trans-S-label led to the detection of proteins secreted by Müller glia cells and by photoreceptors analyzed likewise. Proteins identified include alpha-1-antitrypsin, steroid monooxy-genase, collagens, clusterin, matrix metalloproteinase-2, tropomyosins and thrombospondin 1. 4: Identification of autoantigens for uveitis: Uveitis in horses is an autoimmune disease affecting the retina, as well as other parts of the eye. The identification of autoantigens triggering this disease is crucial for rational treatment. A technique using 2-D Gel separation, blotting, subsequent colloid gold staining and exposure of the blot to the affected horse’s serum led to identification of four autoantigens. Conclusion: Fields of applications for proteomic analyses of the retina but also some of their present limitations will be discussed.

Keywords: retina • protein purification and characterization • gene/expression 
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