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Effects of crystallin-β-b2 on stressed RPE in vitro and in vivo

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Abstract

Background

Crystallins are thought to play a cytoprotective role in conditions of cellular stress. The aim of this study was to determine the effects of crystallin-β-b2 (cryβ-b2) and crystallin-β-b3 (cryβ-b3) on ARPE-19 cells in vitro and on the retinal pigment epithelium (RPE) in vivo.

Methods

The influence of cryβ-b2 and cryβ-b3 on the viability, proliferation and dying of ARPE-19 was measured by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium assay, bromo-2-deoxyuridine assay and life/death assay. The expressions of cryβ-b2, cryβ-b3, glial-derived neurotrophic factor (GDNF), and galectin-3 (Gal-3) in ARPE-19 cells were evaluated using immunohistochemistry (IHC), Western blotting (WB) and real-time-quantitative-PCR (qRT-PCR). To evaluate the response of cryβ-b2 and cryβ-b3 to stressed ARPE-19 cells, the cells were exposed to UV-light. In a rat model, cryβ-b2-expressing neural progenitor cells (cryβ-b2-NPCs) were injected intravitreally after retinal stress induced by optic nerve axotomy to examine whether they influence the RPE. Protein expression was examined 2 and 4 weeks postsurgery using IHC and WB.

Results

Detectable alterations of GDNF, and Gal-3 were found in ARPE-19 cells upon exposure to UV light. Adding the crystallins to the medium promoted proliferation and increased viability of ARPE-19 cells in vitro. The obtained data support the view that these crystallins possess epithelioprotective properties. Likewise, in vivo, intravitreally injected cryβ-b2 and transplanted cryβ-b2-NPCs protected RPE from indirectly induced stress.

Conclusions

The data suggest that the RPE response to retinal ganglion cell denegeration is mediated via crystallins, which may thus be used therapeutically.

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Acknowledgments

The authors thank M. Wissing and M. Langkamp-Flock for their skillful technical assistance, and M. Reis for typing the manuscript. The work was supported by a DFG grant to S.T. (Th 386/18) and by IZKF grants to M.R.R.B. and S.T. The authors thank English Science Editing (ESE) for native linguistic editing of the manuscript.

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Authors and Affiliations

  1. Institute of Experimental Ophthalmology, School of Medicine, Westfalian Wilhelms-University Münster, Albert-Schweitzer-Campus 1, D15, 48149, Münster, Germany

    Michael R. R. Böhm, Harutyun Melkonyan, Patrick Oellers & Solon Thanos

  2. Interdisciplinary Centre for Clinical Research (IZKF), School of Medicine, Westfalian Wilhelms-University Münster, Albert-Schweitzer-Campus 1, D15, 48149, Münster, Germany

    Michael R. R. Böhm & Solon Thanos

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  1. Michael R. R. Böhm
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  2. Harutyun Melkonyan
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Correspondence to Michael R. R. Böhm.

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Oral presentation at the Congress of the German Society of Ophthalmology (DOG) 2011

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Böhm, M.R.R., Melkonyan, H., Oellers, P. et al. Effects of crystallin-β-b2 on stressed RPE in vitro and in vivo. Graefes Arch Clin Exp Ophthalmol 251, 63–79 (2013). https://doi.org/10.1007/s00417-012-2157-7

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  • DOI: https://doi.org/10.1007/s00417-012-2157-7

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