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Functional regeneration of respiratory pathways after spinal cord injury

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Abstract

Spinal cord injuries often occur at the cervical level above the phrenic motor pools, which innervate the diaphragm. The effects of impaired breathing are a leading cause of death from spinal cord injuries, underscoring the importance of developing strategies to restore respiratory activity. Here we show that, after cervical spinal cord injury, the expression of chondroitin sulphate proteoglycans (CSPGs) associated with the perineuronal net (PNN) is upregulated around the phrenic motor neurons. Digestion of these potently inhibitory extracellular matrix molecules with chondroitinase ABC (denoted ChABC) could, by itself, promote the plasticity of tracts that were spared and restore limited activity to the paralysed diaphragm. However, when combined with a peripheral nerve autograft, ChABC treatment resulted in lengthy regeneration of serotonin-containing axons and other bulbospinal fibres and remarkable recovery of diaphragmatic function. After recovery and initial transection of the graft bridge, there was an unusual, overall increase in tonic electromyographic activity of the diaphragm, suggesting that considerable remodelling of the spinal cord circuitry occurs after regeneration. This increase was followed by complete elimination of the restored activity, proving that regeneration is crucial for the return of function. Overall, these experiments present a way to markedly restore the function of a single muscle after debilitating trauma to the central nervous system, through both promoting the plasticity of spared tracts and regenerating essential pathways.

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Figure 1: C2 hemisection results in CSPG production.
Figure 2: A PNG enhances diaphragmatic EMG activity.
Figure 3: ChABC treatment promotes axon regeneration.
Figure 4: PNG transection initially increases tonic diaphragmatic EMG activity then completely eliminates activity.

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Acknowledgements

This project was funded by The Christopher and Dana Reeve Foundation (W.J.A.), the International Spinal Research Trust (W.J.A.), the National Institute of Neurological Disorders and Stroke, grants NS25713 and NS060767 (J.S.), and the National Heart, Lung and Blood Institute, grant HL080318 (T.E.D.). We offer special thanks to the Brumagin Memorial Fund (J.S.) and the Ellen Becker Neuroscience Regenerative Medicine Research Fund (J.S.). We also thank J. D. Houle and V. J. Tom for teaching us the peripheral nerve grafting technique.

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Authors and Affiliations

  1. Department of Neurosciences, Case Western Reserve University School of Medicine, 2109 Adelbert Road, Cleveland, 44106, Ohio, USA

    Warren J. Alilain, Kevin P. Horn, Hongmei Hu, Thomas E. Dick & Jerry Silver

  2. Division of Pulmonary, Department of Medicine, Critical Care, and Sleep Medicine, Case Western Reserve University School of Medicine, Cleveland, 44106, Ohio, USA

    Thomas E. Dick

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  1. Warren J. Alilain
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  2. Kevin P. Horn
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Contributions

W.J.A. performed the surgeries, recorded and analysed the electrophysiological data, and completed the immunocytochemical, tracing and histological experiments. K.P.H. helped with the immunocytochemical detection and quantification. H.H. assisted with the animal care, surgeries and data quantification, as well as performed all of the tissue processing. T.E.D. gave technical guidance and helped discuss the results. W.J.A. and J.S. designed all of the studies, analysed the data and wrote the paper. All authors discussed the data and helped to prepare the manuscript.

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Correspondence to Warren J. Alilain or Jerry Silver.

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The authors declare no competing financial interests.

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Alilain, W., Horn, K., Hu, H. et al. Functional regeneration of respiratory pathways after spinal cord injury. Nature 475, 196–200 (2011). https://doi.org/10.1038/nature10199

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