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Salamanders in Regeneration Research pp 115–125Cite as

High-Efficiency Electroporation of the Spinal Cord in Larval Axolotl

Part of the Methods in Molecular Biology book series (MIMB,volume 1290)

Abstract

Axolotls are well known for their remarkable ability to regenerate complex body parts and structures throughout life, including the entire limb and tail. Particularly fascinating is their ability to regenerate a fully functional spinal cord after losing the tail. Electroporation of DNA plasmids or morpholinos is a valuable tool to gain mechanistic insight into the cellular and molecular basis of regeneration. It provides among other advantages a simple and fast method to test gene function in a temporally and spatially controlled manner. Some classic drawbacks of the method, such as low transfection efficiency and damage to the tissue, had hindered our understanding of the contribution of different signaling pathways to regeneration. Here, we describe a comprehensive protocol for electroporation of the axolotl spinal cord that overcomes this limitations using a combination of high-voltage and short-length pulses followed by lower-voltage and longer-length pulses. Our approach yields highly efficient transfection of spinal cord cells with minimal tissue damage, which now allows the molecular dissection of spinal cord regeneration.

Key words

  • Electroporation
  • Spinal cord
  • Regeneration
  • Axolotl

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  • DOI: 10.1007/978-1-4939-2495-0_9
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Author information

Authors and Affiliations

  1. DFG Center for Regenerative Therapies TU Dresden (CRTD), Technische Universität Dresden, Fetscherstrasse 105, 01307, Dresden, Germany

    Aida Rodrigo Albors & Elly M. Tanaka Ph.D.

Authors
  1. Aida Rodrigo Albors
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  2. Elly M. Tanaka Ph.D.
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Corresponding author

Correspondence to Elly M. Tanaka Ph.D. .

Editor information

Editors and Affiliations

  1. Institute of Structural and Molecular Biology, Division of Biosciences, University College London, London, United Kingdom

    Anoop Kumar

  2. Department of Cell and Molecular Biology, Karolinska Institute, Stockholm, Sweden

    András Simon

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Albors, A.R., Tanaka, E.M. (2015). High-Efficiency Electroporation of the Spinal Cord in Larval Axolotl. In: Kumar, A., Simon, A. (eds) Salamanders in Regeneration Research. Methods in Molecular Biology, vol 1290. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2495-0_9

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  • DOI: https://doi.org/10.1007/978-1-4939-2495-0_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2494-3

  • Online ISBN: 978-1-4939-2495-0

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