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High-Efficiency Electroporation of the Spinal Cord in Larval Axolotl

  • Aida Rodrigo Albors
  • Elly M. Tanaka
Protocol
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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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.DFG Center for Regenerative Therapies TU Dresden (CRTD)Technische Universität DresdenDresdenGermany

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