Assessing Axonal Degeneration in Embryonic Dorsal Root Ganglion Neurons In Vitro

Methods Mol Biol. 2020;2143:41-54. doi: 10.1007/978-1-0716-0585-1_4.

Abstract

The molecular players regulating the axon degeneration pathway have been identified using in vitro experimental models. Here, we describe an in vitro assay to assess the axonal fragmentation induced by mechanical injury to axons in cultured mouse embryonic dorsal root ganglion (DRG) neurons. DRG neurons are pseudounipolar and therefore suitable for an assay of axonal degeneration after injury. In addition, the time course of the axonal fragmentation is stereotyped, enabling the identification of reagents that either expedite or impede the degeneration process. With an image-based quantification method, the in vitro degeneration assay can be utilized as a platform supporting high-throughput screens for pharmacological or genetic reagents delaying axon degeneration.

Keywords: Axon degeneration; Axotomy; Dorsal root ganglion; Image-based screening; In vitro assay; Neurodegeneration.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Axons / physiology*
  • Axotomy
  • Cell Culture Techniques / methods
  • Cells, Cultured
  • Dissection / methods
  • Ganglia, Spinal / cytology*
  • Ganglia, Spinal / embryology
  • Green Fluorescent Proteins / analysis
  • Image Processing, Computer-Assisted / methods
  • Mice
  • Microscopy, Fluorescence / methods
  • Microscopy, Phase-Contrast / methods
  • Primary Cell Culture
  • Sensory Receptor Cells / physiology
  • Sensory Receptor Cells / ultrastructure
  • Wallerian Degeneration / physiopathology*

Substances

  • Green Fluorescent Proteins