Abstract
Sensory neurons from dorsal root ganglion efficiently regenerate after peripheral nerve injuries. These neurons are widely used as a model system to study degenerative mechanisms of the soma and axons, as well as regenerative axonal growth in the peripheral nervous system. This chapter describes techniques associated to the study of axonal degeneration and regeneration using explant cultures of dorsal root ganglion sensory neurons in vitro in the presence or absence of Schwann cells. Schwann cells are extremely important due to their involvement in tissue clearance during axonal degeneration as well as their known pro-regenerative effect during regeneration in the peripheral nervous system. We describe methods to induce and study axonal degeneration triggered by axotomy (mechanical separation of the axon from its soma) and treatment with vinblastine (which blocks axonal transport), which constitute clinically relevant mechanical and toxic models of axonal degeneration. In addition, we describe three different methods to evaluate axonal regeneration using quantitative methods. These protocols constitute a valuable tool to analyze in vitro mechanisms associated to axonal degeneration and regeneration of sensory neurons and the role of Schwann cells in these processes.
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Acknowledgments
We wish to thank all the members of the Court Lab for their contributions to this protocol. This work was supported by Center for Integrative Biology, Universidad Mayor, FONDECYT-1150766, Geroscience Center for Brain Health and Metabolism (FONDAP-15150012), Ring Initiative ACT1109, and Canada-Israel Health Research initiative, jointly Funded by the Canadian Institutes of Health Research; the Israel Science Foundation; the International Development Research Centre, Canada; and the Azrieli Foundation, Canada.
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López-Leal, R., Diaz, P., Court, F.A. (2018). In Vitro Analysis of the Role of Schwann Cells on Axonal Degeneration and Regeneration Using Sensory Neurons from Dorsal Root Ganglia. In: Monje, P., Kim, H. (eds) Schwann Cells. Methods in Molecular Biology, vol 1739. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7649-2_16
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DOI: https://doi.org/10.1007/978-1-4939-7649-2_16
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