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Home / Equine Research Bank / Int J Mol Sci / Profound re-organization of cell surface proteome in equine ...
International journal of molecular sciences2012; 13(11); 14053-14072; doi: 10.3390/ijms131114053

Profound re-organization of cell surface proteome in equine retinal pigment epithelial cells in response to in vitro culturing.

Abstract: The purpose of this study was to characterize the cell surface proteome of native compared to cultured equine retinal pigment epithelium (RPE) cells. The RPE plays an essential role in visual function and represents the outer blood-retinal barrier. We are investigating immunopathomechanisms of equine recurrent uveitis, an autoimmune inflammatory disease in horses leading to breakdown of the outer blood-retinal barrier and influx of autoreactive T-cells into affected horses' vitrei. Cell surface proteins of native and cultured RPE cells from eye-healthy horses were captured by biotinylation, analyzed by high resolution mass spectrometry coupled to liquid chromatography (LC MS/MS), and the most interesting candidates were validated by PCR, immunoblotting and immunocytochemistry. A total of 112 proteins were identified, of which 84% were cell surface membrane proteins. Twenty-three of these proteins were concurrently expressed by both cell states, 28 proteins exclusively by native RPE cells. Among the latter were two RPE markers with highly specialized RPE functions: cellular retinaldehyde-binding protein (CRALBP) and retinal pigment epithelium-specific protein 65kDa (RPE65). Furthermore, 61 proteins were only expressed by cultured RPE cells and absent in native cells. As we believe that initiating events, leading to the breakdown of the outer blood-retinal barrier, take place at the cell surface of RPE cells as a particularly exposed barrier structure, this differential characterization of cell surface proteomes of native and cultured equine RPE cells is a prerequisite for future studies.
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Publication Date: 2012-10-31 PubMed ID: 23203049PubMed Central: PMC3509565DOI: 10.3390/ijms131114053Google Scholar: Lookup
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  • Journal Article
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  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research paper focuses on studying the differences in the cell surface proteins of native and cultured retinal pigment epithelium (RPE) cells in horses, in order to understand the pathomechanisms of equine recurrent uveitis, an autoimmune disease in horses causing inflammation and breakdown of outer blood-retinal barrier.

Research Objective

  • The goal of this study was to investigate the cell surface proteome, which is a complete set of proteins expressed by the genome at the cell surface, of native and cultured equine RPE cells. Information about these proteins may provide insights into the mechanisms of equine recurrent uveitis, a disease known to cause damage to the outer blood-retinal barrier in horses.

Research Methodology

  • RPE cells from healthy horse eyes, both native and cultured, had their cell surface proteins captured through a process called biotinylation. Biotinylation is a biochemical technique that chemically attaches biotin to proteins, DNA, and other molecules.
  • These proteins were then analysed via high resolution mass spectrometry. Mass spectrometry helps in identifying proteins and provide information on their structure and chemical properties.
  • The most interesting candidates amongst these proteins were further validated using methods like PCR (Polymerase Chain Reaction), immunoblotting, and immunocytochemistry.

Research Findings

  • The research identified a total of 112 proteins, 84% of which were cell surface proteins. Twenty-three proteins were expressed by both native and cultured cells.
  • Among the proteins expressed exclusively in native RPE cells, two particular markers – cellular retinaldehyde-binding protein (CRALBP) and retinal pigment epithelium-specific protein 65kDa (RPE65) – play critical roles in RPE function.
  • Interestingly, there were 61 proteins that were only expressed in cultured RPE cells and were completely absent in native cells.

Implication of the Research

  • The research provides valuable information about the differential behaviour and protein expression of native and cultured RPE cells, which is fundamental to understanding the disease mechanisms of equine recurrent uveitis.
  • The researchers believe that the triggers leading to the breakdown of the outer blood-retinal barrier occur at the cell surface of RPE cells, highlighting the importance of this research for future studies.

Cite This Article

APA
Szober CM, Hauck SM, Euler KN, Fröhlich KJ, Alge-Priglinger C, Ueffing M, Deeg CA. (2012). Profound re-organization of cell surface proteome in equine retinal pigment epithelial cells in response to in vitro culturing. Int J Mol Sci, 13(11), 14053-14072. https://doi.org/10.3390/ijms131114053

Publication

International journal of molecular sciences
ISSN: 1422-0067
NlmUniqueID: 101092791
Country: Switzerland
Language: English
Volume: 13
Issue: 11
Pages: 14053-14072

Researcher Affiliations

Szober, Christoph M
  • Institute of Animal Physiology, Department of Veterinary Sciences, Ludwig-Maximilians-University Munich, D-80539 Munich, Germany. deeg@tiph.vetmed.uni-muenchen.de.
Hauck, Stefanie M
    Euler, Kerstin N
      Fröhlich, Kristina J H
        Alge-Priglinger, Claudia
          Ueffing, Marius
            Deeg, Cornelia A

              MeSH Terms

              • Animals
              • Cells, Cultured
              • Epithelial Cells / metabolism
              • Horses
              • Membrane Proteins / metabolism
              • Proteome
              • Proteomics / methods
              • Retinal Pigment Epithelium / metabolism

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