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Na+–K+–2Cl− Cotransport Inhibitor Attenuates Cerebral Edema Following Experimental Stroke via the Perivascular Pool of Aquaporin-4

  • Translational Research
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Abstract

Introduction

The Na+–K+–2Cl− cotransporter localized in the brain vascular endothelium has been shown to be important in the evolution of cerebral edema following experimental stroke. Previous in vivo studies have demonstrated that bumetanide, a selective Na+–K+–2Cl− cotransport inhibitor, attenuates ischemia-evoked cerebral edema. Recently, bumetanide has been shown to also inhibit water permeability via aquaporin-4 (AQP4) expressed in Xenopus laevis oocytes. We tested the hypothesis that the perivascular pool of AQP4 plays a significant role in the anti-edema effect of bumetanide by utilizing wild-type (WT) mice as well as mice with targeted disruption of α-syntrophin (α-Syn−/−) that lack the perivascular pool of AQP4.

Methods

Isoflurane-anesthetized adult male WT C57Bl6 and α-Syn−/− mice were subjected to 90 min middle cerebral artery occlusion (MCAO) followed by 24 or 48 h of reperfusion. Adequacy of MCAO and reperfusion was monitored with laser-Doppler flowmetry over the ipsilateral parietal cortex. Infarct volume (tetrazolium staining), cerebral edema (wet-to-dry ratios), and AQP4 protein expression (immunoblotting) were determined in different treatment groups in separate sets of experiments.

Results

Bumetanide significantly attenuated infarct volume and decreased ipsilateral hemispheric water content in WT mice compared to vehicle treatment. In α-Syn−/− mice, bumetanide treatment had no effect on infarct volume or ischemia-evoked cerebral edema. Bumetanide-treated WT mice had a significant attenuation of AQP4 protein expression at 48 h post-MCAO compared to vehicle-treated WT mice.

Conclusions

These data suggest that bumetanide exerts its neuroprotective and anti-edema effects partly via blockade of the perivascular pool of AQP4 and may have therapeutic potential for ischemic stroke in the clinical setting.

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Acknowledgments

This work was supported by Public Health Service NIH grants NS046379 (AB) and NS33145 (SCF, MEA). The authors thank Stepahnie J. Murphy, D.V.M., Ph.D., and Sarah Mader for maintaining the colony for transgenic mice.

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Authors and Affiliations

  1. Department of Neurology, Oregon Health & Science University, Portland, OR, USA

    Elton R. Migliati & Anish Bhardwaj

  2. Center for Molecular Biology and Neuroscience and Department of Anatomy, Institute of Basic Medical Sciences, University of Oslo, 0317, Oslo, Norway

    Mahmood Amiry-Moghaddam & Ole Petter Ottersen

  3. Department of Physiology and Biophysics, University of Washington, Seattle, WA, 98195, USA

    Stanley C. Froehner & Marvin E. Adams

  4. Department of Anesthesiology and Peri-Operative Medicine, Oregon Health Science University, Portland, OR, USA

    Anish Bhardwaj

  5. Neurological Surgery, Oregon Health Science University, Portland, OR, USA

    Anish Bhardwaj

  6. Department of Neurology, Tufts University School of Medicine, Tufts Medical Center, Box 314, 800 Washington Street, Boston, MA, 02111, USA

    Anish Bhardwaj

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  1. Elton R. Migliati
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  2. Mahmood Amiry-Moghaddam
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  3. Stanley C. Froehner
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  6. Anish Bhardwaj
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Correspondence to Anish Bhardwaj.

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Migliati, E.R., Amiry-Moghaddam, M., Froehner, S.C. et al. Na+–K+–2Cl− Cotransport Inhibitor Attenuates Cerebral Edema Following Experimental Stroke via the Perivascular Pool of Aquaporin-4. Neurocrit Care 13, 123–131 (2010). https://doi.org/10.1007/s12028-010-9376-8

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