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Multiple programmed cell death pathways are involved in N-methyl-N-nitrosourea-induced photoreceptor degeneration

  • Basic Science
  • Published:
Graefe's Archive for Clinical and Experimental Ophthalmology Aims and scope Submit manuscript

Abstract

Purpose

To identify programmed cell death (PCD) pathways involved in N-methyl-N-nitrosourea (MNU)-induced photoreceptor (PR) degeneration.

Methods

Adult C57BL/6 mice received a single MNU i.p. injection (60 mg/kg bodyweight), and were observed over a period of 7 days. Degeneration was visualized by H&E overview staining and electron microscopy. PR cell death was measured by quantifying TUNEL-positive cells in the outer nuclear layer (ONL). Activity measurements of key PCD enzymes (calpain, caspases) were used to identify the involved cell death pathways. Furthermore, the expression level of C/EBP homologous protein (CHOP) and glucose-regulated protein 78 (GRP78), key players in endoplasmic reticulum (ER) stress-induced apoptosis, was analyzed using quantitative real-time PCR.

Results

A decrease in ONL thickness and the appearance of apoptotic PR nuclei could be detected beginning 3 days post-injection (PI). This was accompanied by an increase of TUNEL-positive cells. Significant upregulation of activated caspases (3, 9, 12) was found at different time periods after MNU injection. Additionally, several other players of nonconventional PCD pathways were also upregulated. Consequently, calpain activity increased in the ONL, with a maximum on day 7 PI and an upregulation of CHOP and GRP78 expression beginning on day 1 PI was found.

Conclusions

The data indicate that regular apoptosis is the major cause of MNU-induced PR cell death. However, alternative PCD pathways, including ER stress and calpain activation, are also involved. Knowledge about the mechanisms involved in this mouse model of PR degeneration could facilitate the design of putative combinatory therapeutic approaches.

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Acknowledgments

The authors wish to thank Agathe Duda, Monika Kilchenmann, Anelia Schweri-Olac and Beat Haenni for their excellent technical assistance. This work was partly supported by the Fritz Tobler Foundation and the Bern University Research Foundation.

Author information

Authors and Affiliations

  1. Department of Ophthalmology, Inselspital, University of Bern, Freiburgstrasse 14, 3010, Bern, Switzerland

    Miriam Reisenhofer, Jasmin Balmer & Volker Enzmann

  2. Paul Flechsig Institute for Brain Research, University of Leipzig, Leipzig, Germany

    Miriam Reisenhofer

  3. Graduate School for Cellular and Biomedical Sciences, University of Bern, Bern, Switzerland

    Miriam Reisenhofer

  4. Nuffield Laboratory of Ophthalmology, Department of Clinical Neurosciences, University of Oxford, Oxford, UK

    Jasmin Balmer

  5. Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    Rahel Zulliger

Authors
  1. Miriam Reisenhofer
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  2. Jasmin Balmer
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  4. Volker Enzmann
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Corresponding author

Correspondence to Volker Enzmann.

Additional information

Miriam Reisenhofer and Jasmin Balmer contributed equally to the project.

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Reisenhofer, M., Balmer, J., Zulliger, R. et al. Multiple programmed cell death pathways are involved in N-methyl-N-nitrosourea-induced photoreceptor degeneration. Graefes Arch Clin Exp Ophthalmol 253, 721–731 (2015). https://doi.org/10.1007/s00417-014-2906-x

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  • DOI: https://doi.org/10.1007/s00417-014-2906-x

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