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Traumatic brain injury-induced axonal phenotypes react differently to treatment

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Abstract

Injured axons with distinct morphologies have been found following mild traumatic brain injury (mTBI), although it is currently unclear whether they reflect varied responses to the injury or represent different stages of progressing pathology. This complicates evaluation of therapeutic interventions targeting axonal injury. To address this issue, we assessed axonal injury over time within a well-defined axonal population, while also evaluating mitochondrial permeability transition as a therapeutic target. We utilized mice expressing yellow fluorescent protein (YFP) in cortical neurons which were crossed with mice which lacked Cyclophilin D (CypD), a positive regulator of mitochondrial permeability transition pore opening. Their offspring were subjected to mTBI and the ensuing axonal injury was assessed using YFP expression and amyloid precursor protein (APP) immunohistochemistry, visualized by confocal and electron microscopy. YFP+ axons initially developed a single, APP+, focal swelling (proximal bulb) which progressed to axotomy. Disconnected axonal segments developed either a single bulb (distal bulb) or multiple bulbs (varicosities), which were APP− and whose ultrastructure was consistent with ongoing Wallerian degeneration. CypD knock-out failed to reduce proximal bulb formation but decreased the number of distal bulbs and varicosities, as well as a population of small, APP+, callosal bulbs not associated with YFP+ axons. The observation that YFP+ axons contain several pathological morphologies points to the complexity of traumatic axonal injury. The fact that CypD knock-out reduced some, but not all, subtypes highlights the need to appropriately characterize injured axons when evaluating potential neuroprotective strategies.

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Acknowledgments

The authors thank Carol Davis, Susan Walker and Jesse Sims for invaluable technical assistance, Scott Henderson and Frances White for sharing their expertise in confocal microscopy, Audrey Lafrenaye, Vishal Patel and Michal Vascak for scientific discussions and comments on this manuscript as well as Michael Forte and Paolo Bernardi for generating and providing the CypD KO mice. This work was funded by NIH grants NS077675 and NS047463.

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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Authors and Affiliations

  1. Department of Anatomy and Neurobiology, Medical College of Virginia Campus of Virginia Commonwealth University, Post Office Box 980709, Richmond, VA, 23298-0709, USA

    Anders Hånell, John E. Greer, Melissa J. McGinn & John T. Povlishock

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  1. Anders Hånell
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  2. John E. Greer
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  3. Melissa J. McGinn
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  4. John T. Povlishock
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Correspondence to John T. Povlishock.

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Hånell, A., Greer, J.E., McGinn, M.J. et al. Traumatic brain injury-induced axonal phenotypes react differently to treatment. Acta Neuropathol 129, 317–332 (2015). https://doi.org/10.1007/s00401-014-1376-x

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  • DOI: https://doi.org/10.1007/s00401-014-1376-x

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