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Proteomic profiling reveals crucial retinal protein alterations in the early phase of an experimental glaucoma model

  • Basic Science
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Abstract

Purpose

Clinical glaucoma is difficult to assess in terms of molecular pathophysiology, prompting studies in experimental models of glaucoma. The purpose of this study was to investigate quantitative changes in retinal protein expression at the onset of experimental glaucoma in rats. Analyzing the proteome provides a suitable tool to decipher the pathophysiological processes in glaucomatous degeneration.

Methods

Thermic cauterization of episcleral veins was utilized to elevate the intraocular pressure in Sprague Dawley rats. Morphological changes were surveyed on a cellular level with a staining of Brn3a-positive cells. The retinal nerve fiber layer was investigated using optical coherence tomography (OCT, Heidelberg Engineering) and the optic nerve was analyzed by an axonal grading system. Mass spectrometry-featured quantitative proteomics and immunohistochemical staining was used to identify specifically altered proteins in the course of intraocular pressure elevation and initial neurodegeneration. Proteomic data were further analyzed with Ingenuity Pathway Analysis and Cytoscape to analyze further molecular associations.

Results

The intraocular pressure rose significantly (p < 0.001) for the follow-up period of 3 weeks after which animals were sacrificed. Eyes exposed to an elevated intraocular pressure showed an initial decrease of retinal ganglion cells, retinal nerve fiber layer (p < 0.05) and an impairment of the optic nerve (p < 0.01). Mass spectrometry led to the identification and quantification of 931 retinal proteins, whereas 32 were considerably altered. Bioinformatics-assisted clustering revealed that a majority of these proteins are functionally associated with cell differentiation, apoptosis and stress response. The creation of an interactive protein network showed that numerous altered proteins are connected regarding their cellular function. Protein kinase b, mitogen-activated protein kinase 1 and the NF-κB complex seem to be essential molecules in this context.

Conclusions

In conclusion, these results provide further lines of evidence that substantial molecular changes occur at the onset of the disease, identifying potential key players, which might be useful as biomarkers for diagnostics and development of medical treatment in the future.

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Authors and Affiliations

  1. Experimental Ophthalmology, Department of Ophthalmology, University Medical Center, Langenbeckstrasse 1, 55131, Mainz, Germany

    Fabian Anders, Julia Teister, Sebstian Funke, Norbert Pfeiffer, Franz Grus & Verena Prokosch

  2. University Eye Hospital Mainz, School of Medicine, Langenbeckstrasse 1, 55131, Mainz, Germany

    Norbert Pfeiffer & Verena Prokosch

  3. Department of Experimental Ophthalmology, University Medical Center, Domagkstraße 15, 48149, Münster, Germany

    Thanos Solon

Authors
  1. Fabian Anders
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  2. Julia Teister
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Correspondence to Verena Prokosch.

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Funding

The Deutsche Forschungsgemeinschaft (DFG) provided financial support in the form of a grant for this study (PR1569/1–1). The sponsor had no role in the design or conduct of this research.

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All authors certify that they have no affiliations with or involvement in any organization or entity with any financial interest (such as honoraria; educational grants; participation in speakers’ bureaus; membership, employment, consultancies, stock ownership, or other equity interest; and expert testimony or patent-licensing arrangements), or non-financial interest (such as personal or professional relationships, affiliations, knowledge or beliefs) in the subject matter or materials discussed in this manuscript.

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Ethical approval: All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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Anders, F., Teister, J., Funke, S. et al. Proteomic profiling reveals crucial retinal protein alterations in the early phase of an experimental glaucoma model. Graefes Arch Clin Exp Ophthalmol 255, 1395–1407 (2017). https://doi.org/10.1007/s00417-017-3678-x

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  • DOI: https://doi.org/10.1007/s00417-017-3678-x

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