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Oxidative stress triggers neuronal caspase-independent death: Endonuclease G involvement in programmed cell death-type III

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Abstract

To characterize neuronal death, primary cortical neurons (C57/Black 6 J mice) were exposed to hydrogen peroxide (H2O2) and staurosporine. Both caused cell shrinkage, nuclear condensation, DNA fragmentation and loss of plasma membrane integrity. Neither treatment induced caspase-7 activity, but caspase-3 was activated by staurosporine but not H2O2. Each treatment caused redistribution from mitochondria of both endonuclease G (Endo G) and cytochrome c. Neurons knocked down for Endo G expression using siRNA showed reduction in both nuclear condensation and DNA fragmentation after treatment with H2O2, but not staurosporine. Endo G suppression protected cells against H2O2-induced cell death, while staurosporine-induced death was merely delayed. We conclude that staurosporine induces apoptosis in these neurons, but severe oxidative stress leads to Endo G-dependent death, in the absence of caspase activation (programmed cell death-type III). Therefore, oxidative stress triggers in neurons a form of necrosis that is a systematic cellular response subject to molecular regulation.

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Acknowledgments

We thank Ms Irene Hatzinisiriou for assistance with confocal microscopy and gel scanning, and Dr. Danielle Smith for technical advice. This work was supported by the National Health and Medical Research Council (Australia).

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  1. Department of Biochemistry and Molecular Biology, Monash University, Building 13D, Clayton Campus, Clayton, VIC, 3800, Australia

    Gavin C. Higgins & Phillip Nagley

  2. Florey Neuroscience Institutes and Department of Pharmacology, University of Melbourne, Parkville, VIC, Australia

    Philip M. Beart

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  1. Gavin C. Higgins
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Correspondence to Phillip Nagley.

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Higgins, G.C., Beart, P.M. & Nagley, P. Oxidative stress triggers neuronal caspase-independent death: Endonuclease G involvement in programmed cell death-type III. Cell. Mol. Life Sci. 66, 2773–2787 (2009). https://doi.org/10.1007/s00018-009-0079-2

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