Skip to main content

Functional and Molecular Characterization of a Novel Traumatic Peripheral Nerve–Muscle Injury Model

NeuroMolecular Medicine volume 19, pages 357–374 (2017)Cite this article

Abstract

Traumatic injuries to human peripheral nerves are frequently associated with damage to nerve surrounding tissues including muscles and blood vessels. Currently, most rodent models of peripheral nerve injuries (e.g., facial or sciatic nerve) employ surgical nerve transection with scissors or scalpels. However, such an isolated surgical nerve injury only mildly damages neighboring tissues and weakly activates an immune response. In order to provide a rodent nerve injury model accounting for such nerve-associated tissue damage and immune cell activation, we developed a drop tower-based facial nerve trauma model in mice. We compare nerve regeneration in this novel peripheral nerve trauma model with the established surgical nerve injury along several parameters. These include gene expression, histological and functional facial motoneuron (FMN) regeneration, facial nerve degeneration, immune cell activation and muscle damage. Regeneration-associated genes (RAGs; e.g., Atf3) were strongly induced in FMNs subjected to traumatic and surgical injury. Regeneration of FMNs and functional recovery of whisker movement were faster in traumatic versus complete surgical injury, thus cutting down experimentation time. Wallerian degeneration of distal nerve stumps was readily observed in this novel trauma injury model. Importantly, drop tower-inflicted facial nerve injury resulted in muscle damage, activation of muscle satellite cell markers (PAX7) and pronounced infiltration of immune cells to the injury site only in this model but not upon surgical nerve transection. Thus, we provide a novel rodent PNS trauma model that can be easily adopted to other PNS nerves such as the sciatic nerve. Since this nerve trauma model replicates multiple tissue damage frequently encountered in clinical routine, it will be well suited to identify molecular and cellular mechanisms of PNS nerve repair in wild-type and genetically modified rodents.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

References

  • Albert-Weissenberger, C., & Siren, A. L. (2010). Experimental traumatic brain injury. Experimental & Translational Stroke Medicine, 2, 16.

    Article  Google Scholar 

  • Banks, C. A., Knox, C., Hunter, D. A., Mackinnon, S. E., Hohman, M. H., & Hadlock, T. A. (2015). Long-term functional recovery after facial nerve transection and repair in the rat. Journal of Reconstructive Microsurgery, 31, 210–216.

    Article  PubMed  PubMed Central  Google Scholar 

  • Campbell, W. W. (2008). Evaluation and management of peripheral nerve injury. Clinical Neurophysiology: Official Journal of the International Federation of Clinical Neurophysiology, 119, 1951–1965.

    Article  Google Scholar 

  • Chandran, V., Coppola, G., Nawabi, H., Omura, T., Versano, R., Huebner, E. A., et al. (2016). A systems-level analysis of the peripheral nerve intrinsic axonal growth program. Neuron, 89, 956–970.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Chen, P., Piao, X., & Bonaldo, P. (2015). Role of macrophages in Wallerian degeneration and axonal regeneration after peripheral nerve injury. Acta Neuropathologica, 130, 605–618.

    CAS  Article  PubMed  Google Scholar 

  • Dalamagkas, K., Tsintou, M., & Seifalian, A. (2016). Advances in peripheral nervous system regenerative therapeutic strategies: A biomaterials approach. Materials Science & Engineering C, Materials for Biological Applications, 65, 425–432.

    CAS  Article  Google Scholar 

  • Di Giovanni, S., Knights, C. D., Rao, M., Yakovlev, A., Beers, J., Catania, J., et al. (2006). The tumor suppressor protein p53 is required for neurite outgrowth and axon regeneration. The EMBO Journal, 25, 4084–4096.

    Article  PubMed  PubMed Central  Google Scholar 

  • Faroni, A., Mobasseri, S. A., Kingham, P. J., & Reid, A. J. (2015). Peripheral nerve regeneration: Experimental strategies and future perspectives. Advanced Drug Delivery Reviews, 82–83, 160–167.

    Article  PubMed  Google Scholar 

  • Fernandez-Verdejo, R., Vanwynsberghe, A. M., Essaghir, A., Demoulin, J. B., Hai, T., Deldicque, L., & Francaux, M. (2017). Activating transcription factor 3 attenuates chemokine and cytokine expression in mouse skeletal muscle after exercise and facilitates molecular adaptation to endurance training. FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology, 31(2), 840–851.

    CAS  Article  Google Scholar 

  • Flierl, M. A., Stahel, P. F., Beauchamp, K. M., Morgan, S. J., Smith, W. R., & Shohami, E. (2009). Mouse closed head injury model induced by a weight-drop device. Nature Protocols, 4, 1328–1337.

    CAS  Article  PubMed  Google Scholar 

  • Fox, I. K., & Mackinnon, S. E. (2011). Adult peripheral nerve disorders: Nerve entrapment, repair, transfer, and brachial plexus disorders. Plastic and Reconstructive Surgery, 127, 105e–118e.

    Article  PubMed  Google Scholar 

  • Gaudet, A. D., Popovich, P. G., & Ramer, M. S. (2011). Wallerian degeneration: Gaining perspective on inflammatory events after peripheral nerve injury. Journal of Neuroinflammation, 8, 110.

    Article  PubMed  PubMed Central  Google Scholar 

  • Gey, M., Wanner, R., Schilling, C., Pedro, M. T., Sinske, D., & Knoll, B. (2016). Atf3 mutant mice show reduced axon regeneration and impaired regeneration-associated gene induction after peripheral nerve injury. Open Biology, 6, 160091.

    Article  PubMed  PubMed Central  Google Scholar 

  • Gordon, T. (2016). Nerve regeneration: Understanding biology and its influence on return of function after nerve transfers. Hand Clinics, 32, 103–117.

    Article  PubMed  Google Scholar 

  • Gordon, T., & Borschel, G. H. (2017). The use of the rat as a model for studying peripheral nerve regeneration and sprouting after complete and partial nerve injuries. Experimental Neurology, 287, 331–347.

    CAS  Article  PubMed  Google Scholar 

  • Griffin, J. W., Pan, B., Polley, M. A., Hoffman, P. N., & Farah, M. H. (2010). Measuring nerve regeneration in the mouse. Experimental Neurology, 223, 60–71.

    CAS  Article  PubMed  Google Scholar 

  • Grinsell, D., & Keating, C. P. (2014). Peripheral nerve reconstruction after injury: A review of clinical and experimental therapies. BioMed Research International, 2014, 698256.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Grosheva, M., Guntinas-Lichius, O., Angelova, S. K., Kuerten, S., Alvanou, A., Streppel, M., et al. (2008). Local stabilization of microtubule assembly improves recovery of facial nerve function after repair. Experimental Neurology, 209, 131–144.

    CAS  Article  PubMed  Google Scholar 

  • Guntinas-Lichius, O., Irintchev, A., Streppel, M., Lenzen, M., Grosheva, M., Wewetzer, K., et al. (2005). Factors limiting motor recovery after facial nerve transection in the rat: Combined structural and functional analyses. The European Journal of Neuroscience, 21, 391–402.

    Article  PubMed  Google Scholar 

  • Irintchev, A. (2011). Potentials and limitations of peripheral nerve injury models in rodents with particular reference to the femoral nerve. Annals of Anatomy = Anatomischer Anzeiger: Official Organ of the Anatomische Gesellschaft, 193, 276–285.

    Article  Google Scholar 

  • Jessen, K. R., & Mirsky, R. (2008). Negative regulation of myelination: Relevance for development, injury, and demyelinating disease. Glia, 56, 1552–1565.

    Article  PubMed  Google Scholar 

  • Jessen, K. R., Mirsky, R., & Lloyd, A. C. (2015). Schwann cells: Development and role in nerve repair. Cold Spring Harbor Perspectives in Biology, 7, a020487.

    Article  PubMed  PubMed Central  Google Scholar 

  • Kaariainen, M., & Kauhanen, S. (2012). Skeletal muscle injury and repair: The effect of disuse and denervation on muscle and clinical relevance in pedicled and free muscle flaps. Journal of Reconstructive Microsurgery, 28, 581–587.

    Article  PubMed  Google Scholar 

  • Magill, C., Whitlock, E., Solowski, N., & Myckatyn, T. (2008). Transgenic models of nerve repair and nerve regeneration. Neurological Research, 30, 1023–1029.

    Article  PubMed  Google Scholar 

  • Marklund, N., & Hillered, L. (2011). Animal modelling of traumatic brain injury in preclinical drug development: Where do we go from here? British Journal of Pharmacology, 164, 1207–1229.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Moran, L. B., & Graeber, M. B. (2004). The facial nerve axotomy model. Brain Research. Brain Research Reviews, 44, 154–178.

    Article  PubMed  Google Scholar 

  • Muheremu, A., & Ao, Q. (2015). Past, present, and future of nerve conduits in the treatment of peripheral nerve injury. BioMed Research International, 2015, 237507.

    Article  PubMed  PubMed Central  Google Scholar 

  • Navarro, X. (2016). Functional evaluation of peripheral nerve regeneration and target reinnervation in animal models: A critical overview. The European Journal of Neuroscience, 43, 271–286.

    Article  PubMed  Google Scholar 

  • Placheta, E., Wood, M. D., Lafontaine, C., Frey, M., Gordon, T., & Borschel, G. H. (2015). Macroscopic in vivo imaging of facial nerve regeneration in Thy1-GFP rats. JAMA Facial Plastic Surgery, 17, 8–15.

    Article  PubMed  Google Scholar 

  • Post, A., Hoshizaki, T. B., Gilchrist, M. D., Brien, S., Cusimano, M., & Marshall, S. (2015). Traumatic brain injuries: The influence of the direction of impact. Neurosurgery, 76, 81–91.

    Article  PubMed  Google Scholar 

  • Raivich, G., Bohatschek, M., Da Costa, C., Iwata, O., Galiano, M., Hristova, M., et al. (2004). The AP-1 transcription factor c-Jun is required for efficient axonal regeneration. Neuron, 43, 57–67.

    CAS  Article  PubMed  Google Scholar 

  • Relaix, F., & Zammit, P. S. (2012). Satellite cells are essential for skeletal muscle regeneration: The cell on the edge returns centre stage. Development, 139, 2845–2856.

    CAS  Article  PubMed  Google Scholar 

  • Seijffers, R., Mills, C. D., & Woolf, C. J. (2007). ATF3 increases the intrinsic growth state of DRG neurons to enhance peripheral nerve regeneration. The Journal of Neuroscience: The Official Journal of the Society for Neuroscience, 27, 7911–7920.

    CAS  Article  Google Scholar 

  • Seitz, M., Grosheva, M., Skouras, E., Angelova, S. K., Ankerne, J., Jungnickel, J., et al. (2011). Poor functional recovery and muscle polyinnervation after facial nerve injury in fibroblast growth factor-2-/- mice can be improved by manual stimulation of denervated vibrissal muscles. Neuroscience, 182, 241–247.

    CAS  Article  PubMed  Google Scholar 

  • Skouras, E., Ozsoy, U., Sarikcioglu, L., & Angelov, D. N. (2011). Intrinsic and therapeutic factors determining the recovery of motor function after peripheral nerve transection. Annals of anatomy = Anatomischer Anzeiger: Official Organ of the Anatomische Gesellschaft, 193(303), 286–303.

    Article  Google Scholar 

  • Stern, S., Haverkamp, S., Sinske, D., Tedeschi, A., Naumann, U., Di Giovanni, S., et al. (2013). The transcription factor serum response factor stimulates axon regeneration through cytoplasmic localization and cofilin interaction. The Journal of Neuroscience: The Official Journal of the Society for Neuroscience, 33, 18836–18848.

    CAS  Article  Google Scholar 

  • Stern, S., Sinske, D., & Knoll, B. (2012). Serum response factor modulates neuron survival during peripheral axon injury. Journal of Neuroinflammation, 9, 78.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Takeoka, A., Vollenweider, I., Courtine, G., & Arber, S. (2014). Muscle spindle feedback directs locomotor recovery and circuit reorganization after spinal cord injury. Cell, 159, 1626–1639.

    CAS  Article  PubMed  Google Scholar 

  • Tedeschi, A. (2011). Tuning the orchestra: Transcriptional pathways controlling axon regeneration. Frontiers in Molecular Neuroscience, 4, 60.

    PubMed  Google Scholar 

  • van Kesteren, R. E., Mason, M. R., Macgillavry, H. D., Smit, A. B., & Verhaagen, J. (2011). A gene network perspective on axonal regeneration. Frontiers in Molecular Neuroscience, 4, 46.

    PubMed  PubMed Central  Google Scholar 

  • Wood, M. D., & Mackinnon, S. E. (2015). Pathways regulating modality-specific axonal regeneration in peripheral nerve. Experimental Neurology, 265, 171–175.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Xiong, Y., Mahmood, A., & Chopp, M. (2013). Animal models of traumatic brain injury. Nature Reviews Neuroscience, 14, 128–142.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

B.K. is supported by the DFG (Deutsche Forschungsgemeinschaft) through SFB1149 and an Ulm University and Bundeswehrkrankenhaus research initiative (U2.1d E/U2AD/ED002/EF550).

Author information

Affiliations

  1. Institute of Physiological Chemistry, Ulm University, Albert-Einstein-Allee 11, 89081, Ulm, Germany

    Renate Wanner, Manuel Gey & Bernd Knöll

  2. Core Facility Small Animal Imaging, Department of Internal Medicine II, Ulm University, 89081, Ulm, Germany

    Alireza Abaei & Volker Rasche

  3. Institute of Orthopedic Research and Biomechanics, Centre for Trauma Research Ulm, Ulm University Medical Centre, 89081, Ulm, Germany

    Daniela Warnecke, Luisa de Roy & Lutz Dürselen

Authors
  1. Renate Wanner
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Manuel Gey
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Alireza Abaei
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Daniela Warnecke
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Luisa de Roy
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Lutz Dürselen
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Volker Rasche
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Bernd Knöll
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

RW, MG performed and analyzed all experiments. BK conceived the project, supervised all research and has written the manuscript. DW, LdR and LD performed the evaluation of impact force and participated in writing and revising the manuscript. VR and AA performed MRI studies.

Corresponding author

Correspondence to Bernd Knöll.

Ethics declarations

Conflict of interest

The authors declare no competing financial interests.

Electronic Supplementary Material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 2393 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Wanner, R., Gey, M., Abaei, A. et al. Functional and Molecular Characterization of a Novel Traumatic Peripheral Nerve–Muscle Injury Model. Neuromol Med 19, 357–374 (2017). https://doi.org/10.1007/s12017-017-8450-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12017-017-8450-1

Keywords

  • PNS
  • Axon regeneration
  • Facial nerve
  • ATF3
  • Muscle