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Drosophila Laser Axotomy Injury Model to Investigate RNA Repair and Splicing in Axon Regeneration

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Axon Regeneration

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2636))

Abstract

The limited axon regeneration capacity of mature neurons often leads to insufficient functional recovery after damage to the central nervous system (CNS). To promote CNS nerve repair, there is an urgent need to understand the regeneration machinery in order to develop effective clinical therapies. To this aim, we developed a Drosophila sensory neuron injury model and the accompanying behavioral assay to examine axon regeneration competence and functional recovery after injury in the peripheral and central nervous systems. Specifically, we used a two-photon laser to induce axotomy and performed live imaging to assess axon regeneration, combined with the analysis of the thermonociceptive behavior as a readout of functional recovery. Using this model, we found that the RNA 3′-terminal phosphate cyclase (Rtca), which acts as a regulator for RNA repair and splicing, responds to injury-induced cellular stress and impedes axon regeneration after axon breakage. Here we describe how we utilize our Drosophila model to assess the role of Rtca during neuroregeneration.

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Acknowledgments

We thank members of the Song lab for helpful discussions. Y.S. is a recipient of the National Institute of Neurological Disorders and Stroke (NINDS) Pathway to Independence Award. This work was supported by an IDDRC New Program Development Award (CHOP/Penn), an NINDS K99/R00 award (R00NS088211), an NIH grant (1R01NS107392), and a Craig H. Neilsen Foundation research grant to Y.S.

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Authors and Affiliations

  1. Raymond G. Perelman Center for Cellular and Molecular Therapeutics, The Children’s Hospital of Philadelphia, Philadelphia, PA, USA

    Qin Wang, Shannon Trombley, Mahdi Rashidzada & Yuanquan Song

  2. Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA, USA

    Yuanquan Song

Authors
  1. Qin Wang
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  2. Shannon Trombley
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  3. Mahdi Rashidzada
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  4. Yuanquan Song
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Corresponding author

Correspondence to Yuanquan Song .

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Editors and Affiliations

  1. Department of Biology, Appalachian State University, Boone, NC, USA

    Ava J. Udvadia

  2. Office of Technology Development, Medical College of Wisconsin, Milwaukee, WI, USA

    James B. Antczak

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Wang, Q., Trombley, S., Rashidzada, M., Song, Y. (2023). Drosophila Laser Axotomy Injury Model to Investigate RNA Repair and Splicing in Axon Regeneration. In: Udvadia, A.J., Antczak, J.B. (eds) Axon Regeneration. Methods in Molecular Biology, vol 2636. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3012-9_22

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  • DOI: https://doi.org/10.1007/978-1-0716-3012-9_22

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3011-2

  • Online ISBN: 978-1-0716-3012-9

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