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The voltage-gated proton channel Hv1 enhances brain damage from ischemic stroke

Nature Neuroscience volume 15, pages 565–573 (2012)Cite this article

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Abstract

Phagocytic cell NADPH oxidase (NOX) generates reactive oxygen species (ROS) as part of innate immunity. Unfortunately, ischemia can also induce this pathway and inflict damage on native cells. The voltage-gated proton channel Hv1 enables NOX function by compensating cellular loss of electrons with protons. Accordingly, we investigated whether NOX-mediated brain damage in stroke can be inhibited by suppression of Hv1. We found that mouse and human brain microglia, but not neurons or astrocytes, expressed large Hv1-mediated currents. Hv1 was required for NOX-dependent ROS generation in brain microglia in situ and in vivo. Mice lacking Hv1 were protected from NOX-mediated neuronal death and brain damage 24 h after stroke. These results indicate that Hv1-dependent ROS production is responsible for a substantial fraction of brain damage at early time points after ischemic stroke and provide a rationale for Hv1 as a therapeutic target for the treatment of ischemic stroke.

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Figure 1: Voltage-gated proton currents in hippocampal microglia in mouse brain slices.
Figure 2: Hv1 mediates the voltage-gated proton current in mouse brain microglia, but not in neurons or astrocytes.
Figure 3: Hv1 regulates microglial acid extrusion and controls NOX–dependent ROS production in situ.
Figure 4: Hv1−/− mice are protected in ischemic stroke.
Figure 5: Hv1-deficient mice are relatively protected against ischemic neuronal death.
Figure 6: Microglial Hv1 is critical for ROS production after stroke.
Figure 7: Microglial, but not leukocyte, Hv1 is responsible for brain damage after ischemic stroke.

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Acknowledgements

We thank L. Silberstein, W. Yang and M. Kurtev for technical assistance, S. Doctrow (Boston University) for kindly providing us with EUK-207, Y. Kirichok (University of California, San Francisco) for Hv1 antibody, and D. Chaudhuri for critical reading of the manuscript. This work was supported by the US National Institutes of Health (R01 MH090293 to D.E.C. and R01 EY019029 to E.P.F.). L.-J.W. is supported by a Lefler postdoctoral fellowship from Harvard Medical School and a Scientist Development Grant from the American Heart Association (11SDG7340011).

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Author notes

  1. Long-Jun Wu and Gongxiong Wu: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Cardiology, Children's Hospital Boston and Department of Neurobiology, Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts, USA

    Long-Jun Wu & David E Clapham

  2. Research Division, Joslin Diabetes Center, Harvard Medical School, Boston, Massachusetts, USA

    Gongxiong Wu & Edward P Feener

  3. Department of Radiology, Beth Israel Deaconess Medical Center, Boston, Massachusetts, USA

    M Reza Akhavan Sharif

  4. Department of Radiology, Children's Hospital Boston, Boston, Massachusetts, USA

    Amanda Baker & Frederic H Fahey

  5. Department of Laboratory Medicine, Children's Hospital Boston, Boston, Massachusetts, USA

    Yonghui Jia & Hongbo R Luo

  6. Department of Pathology, Harvard Medical School, Boston, Massachusetts, USA

    Yonghui Jia & Hongbo R Luo

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  1. Long-Jun Wu
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Contributions

L.-J.W. performed experiments, including electrophysiology, imaging, immunostaining, cell culture, quantitative RT-PCR, western blot and the mouse stroke models. G.W. performed the mouse stroke models, and western blot, immunostaining, quantitative RT-PCR and cell culture experiments. M.R.A.S. conducted MRI experiments. A.B. conducted PET and computed tomography imaging experiments. Y.J conducted bone marrow transplantation experiments. L.-J.W., G.W., Y.J., F.H.F. and D.E.C. conducted the data analyses. L.-J.W. and D.E.C. wrote the manuscript. L.-J.W., H.R.L., E.P.F. and D.E.C. supervised the project.

Corresponding author

Correspondence to David E Clapham.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–14 (PDF 1238 kb)

Supplementary Movie 1

Time-lapse imaging of increased ROS production in wt and Hv1−/− microglia in brain slices after PMA perfusion (10s/frame) (AVI 4333 kb)

Supplementary Movie 2

Time-lapse imaging of ATP-induced chemotaxis in wt and Hv1−/− microglia in brain slices (1 min/frame) (AVI 3686 kb)

Supplementary Movie 3

MRI images of coronal section of whole brain 24 hours after pMCAO (AVI 1156 kb)

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Wu, LJ., Wu, G., Sharif, M. et al. The voltage-gated proton channel Hv1 enhances brain damage from ischemic stroke. Nat Neurosci 15, 565–573 (2012). https://doi.org/10.1038/nn.3059

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