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N-terminally cleaved Bcl-xL mediates ischemia-induced neuronal death

Nature Neuroscience volume 15, pages 574–580 (2012)Cite this article

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Abstract

Transient global ischemia in rats induces delayed death of hippocampal CA1 neurons. Early events include caspase activation, cleavage of anti-death Bcl-2 family proteins and large mitochondrial channel activity. However, whether these events have a causal role in ischemia-induced neuronal death is unclear. We found that the Bcl-2 and Bcl-xL inhibitor ABT-737, which enhances death of tumor cells, protected rats against neuronal death in a clinically relevant model of brain ischemia. Bcl-xL is prominently expressed in adult neurons and can be cleaved by caspases to generate a pro-death fragment, ΔN-Bcl-xL. We found that ABT-737 administered before or after ischemia inhibited ΔN-Bcl-xL–induced mitochondrial channel activity and neuronal death. To establish a causal role for ΔN-Bcl-xL, we generated knock-in mice expressing a caspase-resistant form of Bcl-xL. The knock-in mice exhibited markedly reduced mitochondrial channel activity and reduced vulnerability to ischemia-induced neuronal death. These findings suggest that truncated Bcl-xL could be a potentially important therapeutic target in ischemic brain injury.

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Figure 1: Treatment with the Bcl-xL inhibitor ABT-737 protects against ischemia-induced neuronal death in the CA1.
Figure 2: The Bcl-xL inhibitor ABT-737 blocks ischemia-induced mitochondrial channel formation.
Figure 3: ABT-737 directly attenuates channel activity in post-ischemic mitochondria.
Figure 4: Pretreatment of animals with ABT-737 attenuates ischemia-induced cleavage of Bcl-xL.
Figure 5: ABT-737 attenuates ΔN61-Bcl-xL–elicited channel activity and cytochrome c release.
Figure 6: ΔN-Bcl-xL induces cell death in hippocampal neurons and Bax−/−; Bak−/− MEFs.
Figure 7: Bcl-xL cleavage-resistant mice are protected against ischemia-induced neuronal death.

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Acknowledgements

We thank A. Latuszek-Barrantes for excellent technical assistance, and J. Hickman, R. Kitsis and L.K. Kaczmarek for insightful scientific discussion and constructive review of the manuscript. We thank C. Kinnally and N. Danial for the gift of Bax−/−; Bak−/− MEFs and the Institut de Recherches Servier for ABT-737. This work was supported by grants from the US National Institutes of Health (NS045876 to E.A.J., NS46742 to R.S.Z. and NS37402 to J.M.H.), a McKnight Foundation Brain Disorders Award (to R.S.Z.) and a generous grant from the F.M. Kirby Foundation (to R.S.Z.). R.S.Z. is the F.M. Kirby Professor of Neural Repair and Protection.

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Authors and Affiliations

  1. Dominick P. Purpura Department of Neuroscience, Albert Einstein College of Medicine, Bronx, New York, USA

    Dimitry Ofengeim, Takahiro Miyawaki, Fabrizio Pontarelli & R Suzanne Zukin

  2. Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

    Ying-bei Chen & J Marie Hardwick

  3. Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut, USA

    Hongmei Li, Silvio Sacchetti, Richard J Flannery, Kambiz N Alavian & Elizabeth A Jonas

  4. Department of Biochemistry and Molecular Biology, Johns Hopkins University School of Public Health, Baltimore, Maryland, USA

    Brian A Roelofs & J Marie Hardwick

  5. Institut de Recherches Servier, Croissy-sur-Seine, France

    John A Hickman

  6. Department of Molecular Microbiology and Immunology, Johns Hopkins University School of Public Health, Baltimore, Maryland, USA

    J Marie Hardwick

  7. Department Neurobiology, Yale University School of Medicine, New Haven, Connecticut, USA

    Elizabeth A Jonas

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Contributions

D.O. performed the experiments, wrote and prepared the manuscript, and made intellectual contributions to the study. Y.-b.C. created the knock-in mouse. T.M., H.L., S.S., R.J.F. and F.P. performed experiments. K.N.A. performed experiments and made intellectual contributions. B.A.R. assisted with knock-in mouse colony preparation. J.A.H. provided intellectual contributions. J.M.H. and R.S.Z. designed experiments, wrote the manuscript and provided intellectual contributions. E.A.J. designed experiments, performed experiments, wrote the manuscript and provided intellectual contributions.

Corresponding authors

Correspondence to J Marie Hardwick, R Suzanne Zukin or Elizabeth A Jonas.

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Ofengeim, D., Chen, Yb., Miyawaki, T. et al. N-terminally cleaved Bcl-xL mediates ischemia-induced neuronal death. Nat Neurosci 15, 574–580 (2012). https://doi.org/10.1038/nn.3054

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