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In vivo imaging of axonal degeneration and regeneration in the injured spinal cord

Nature Medicine volume 11, pages 572–577 (2005)Cite this article

Abstract

The poor response of central axons to transection underlies the bleak prognosis following spinal cord injury. Here, we monitor individual fluorescent axons in the spinal cords of living transgenic mice over several days after spinal injury. We find that within 30 min after trauma, axons die back hundreds of micrometers. This acute form of axonal degeneration is similar in mechanism to the more delayed Wallerian degeneration of the disconnected distal axon, but acute degeneration affects the proximal and distal axon ends equally. In vivo imaging further shows that many axons attempt regeneration within 6–24 h after lesion. This growth response, although robust, seems to fail as a result of the inability of axons to navigate in the proper direction. These results suggest that time-lapse imaging of spinal cord injury may provide a powerful analytical tool for assessing the pathogenesis of spinal cord injury and for evaluating therapies that enhance regeneration.

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Figure 1: Fluorescent labeling of sensory DRG axons in GFP-S mice.
Figure 2: Repetitive in vivo imaging of DRG axons after spinal cord injury.
Figure 3: Imaging acute axonal degeneration in vivo.
Figure 4: Monitoring early axonal outgrowth in vivo.

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Acknowledgements

The authors wish to thank J. Sanes for providing GFP-S mice and support, T.C. Saido for providing the antibody specific for spectrin degradation products, J. Tollett for animal husbandry, J. Lu for writing software, L. Godinho, F. Bareyre and J. Sanes for suggestions and critical reading of the manuscript. This work was supported by grants from the Christopher Reeve Paralysis Foundation (T.M.), the Emmy Noether Program of the Deutsche Forschungsgemeinschaft (T.M., M.K.) and grants from the Swiss National Science Foundation (M.E.S.) and the US National Institutes of Health (J.W.L.).

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Authors and Affiliations

  1. Department of Molecular and Cellular Biology, Harvard University, 7 Divinity Avenue, Cambridge, 02138, Massachusetts, USA

    Martin Kerschensteiner, Jeff W Lichtman & Thomas Misgeld

  2. Department of Neuromorphology, Department of Biology, Brain Research Institute, ETH and University of Zurich, Winterthurer Strasse 190, Zurich, CH-8057, Switzerland

    Martin Kerschensteiner & Martin E Schwab

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  1. Martin Kerschensteiner
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  2. Martin E Schwab
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Correspondence to Thomas Misgeld.

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Supplementary information

Supplementary Fig. 1

Wallerian degeneration. (PDF 81 kb)

Supplementary Fig. 2

Acute axonal degeneration is reduced in WLD-S mice. (PDF 90 kb)

Supplementary Fig. 3

Calpain mediates acute axonal degeneration. (PDF 286 kb)

Supplementary Fig. 4

Neurofilament staining in acute axonal degeneration. (PDF 379 kb)

Supplementary Fig. 5

Nodal sprout during early axonal out-growth. (PDF 88 kb)

Supplementary Fig. 6

Reconstruction of a sprouting DRG neuron. (PDF 120 kb)

Supplementary Movie 1 (AVI 2304 kb)

Supplementary Movie 2 (AVI 2906 kb)

Supplementary Movie 3 (AVI 1161 kb)

Supplementary Movie 4 (AVI 1232 kb)

Supplementary Movie 5 (AVI 1688 kb)

Supplementary Movie 6 (AVI 1770 kb)

Supplementary Movie 7 (AVI 2100 kb)

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Kerschensteiner, M., Schwab, M., Lichtman, J. et al. In vivo imaging of axonal degeneration and regeneration in the injured spinal cord. Nat Med 11, 572–577 (2005). https://doi.org/10.1038/nm1229

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