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Temporal Profiles of Cytoskeletal Protein Loss following Traumatic Axonal Injury in Mice

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Abstract

To examine the time course and relative extent of proteolysis of neurofilament and tubulin proteins after traumatic axonal injury (TAI), anesthetized mice were subjected to optic nerve stretch injury. Immunohistochemistry confirmed neurofilament accumulation within axonal swellings at 4, 24, and 72 h postinjury (n = 4 injured and 2 sham per time point). Immunoblotting of optic nerve homogenates (n = 5 injured and 1 sham at 0.5, 4, 24 or 72 h) revealed calpain-mediated spectrin proteolytic fragments after injury. Protein levels for NF68 progressively decreased from 0.5 h to 24 h postinjury, while NF200 and α-tubulin levels decreased acutely (0.5–4 h), with a secondary decline at 72 h postinjury. These data demonstrate that diffusely distributed TAI is associated not only with a localized accumulation of neurofilament proteins, but also significant decreases in total cytoskeletal protein levels which may be mediated, in part, by calpains. Protection of the axonal cytoskeleton represents a potential therapeutic target for axonal damage associated with injury or neurodegenerative diseases.

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Acknowledgments

This study was supported, in part, by NIH P50-NS08803, R01-NS45131, R01-NS41561 and T32-NS043126. We would like to thank Dr. Juan Wang for assistance with immunoblotting and Heather N. Foozer for assistance with manuscript preparation.

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Authors and Affiliations

  1. Department of Neurosurgery, University of Pennsylvania, Philadelphia, PA, USA

    Gulyeter Serbest

  2. Department of Pharmacology, University of Pennsylvania, Philadelphia, PA, USA

    Robert Siman

  3. Department of Neurobiology & Anatomy, Drexel University College of Medicine, Philadelphia, PA, USA

    Ramesh Raghupathi

  4. Department of Physiology, Spinal Cord & Brain Injury Research Center, University of Kentucky, B367 Biomedical & Biological Sciences Research Bldg (BBSRB), 741 S. Limestone St, Lexington, KY, 40536-0509, USA

    Matthew F. Burkhardt & Kathryn E. Saatman

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  1. Gulyeter Serbest
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  2. Matthew F. Burkhardt
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Correspondence to Kathryn E. Saatman.

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Special issue in honor of Naren Banik.

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Serbest, G., Burkhardt, M.F., Siman, R. et al. Temporal Profiles of Cytoskeletal Protein Loss following Traumatic Axonal Injury in Mice. Neurochem Res 32, 2006–2014 (2007). https://doi.org/10.1007/s11064-007-9318-9

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