Investigating mammalian axon regeneration: In vivo electroporation of adult mouse dorsal root ganglion

Qiao Li, Cheng Qian, Feng Quan Zhou

Research output: Contribution to journalArticlepeer-review

Abstract

Electroporation is an essential non-viral gene transfection approach to introduce DNA plasmids or small RNA molecules into cells. A sensory neuron in the dorsal root ganglion (DRGs) extends a single axon with two branches. One branch goes to the peripheral nerve (peripheral branch), and the other branch enters the spinal cord through the dorsal root (central branch). After the neural injury, the peripheral branch regenerates robustly whereas the central branch does not regenerate. Due to the high regenerative capacity, sensory axon regeneration has been widely used as a model system to study mammalian axon regeneration in both the peripheral nervous system (PNS) and the central nervous system (CNS). Here, we describe a previously established approach protocol to manipulate gene expression in mature sensory neurons in vivo via electroporation. Based on transfection with plasmids or small RNA oligos (siRNAs or microRNAs), the approach allows for both loss-and gain-of-function experiments to study the roles of genes-of-interests or microRNAs in regulation of axon regeneration in vivo. In addition, the manipulation of gene expression in vivo can be controlled both spatially and temporally within a relatively short time course. This model system provides a unique tool to investigate the molecular mechanisms by which mammalian axon regeneration is regulated in vivo.

Original languageEnglish (US)
Article numbere58171
JournalJournal of Visualized Experiments
Volume2018
Issue number139
DOIs
StatePublished - Sep 1 2018

Keywords

  • Axon regeneration
  • Dorsal root ganglion
  • Drg
  • Electroporation
  • Genetic manipulation
  • In vivo
  • Issue 139
  • Neuroscience
  • Non-viral transfection
  • Peripheral nerve injury
  • Sensory neuron

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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