In Vivo Analysis of Glial Immune Responses to Axon Degeneration in Drosophila melanogaster

Methods Mol Biol. 2020:2143:321-338. doi: 10.1007/978-1-0716-0585-1_24.

Abstract

Axon degeneration elicits a range of immune responses from local glial cells, including striking changes in glial gene expression, morphology, and phagocytic activity. Here, we describe a detailed set of protocols to assess discrete components of the glial reaction to axotomy in the adult nervous system of Drosophila melanogaster. These methods allow one to visualize and quantify transcriptional, morphological, and functional responses of glia to degenerating axons in a model system that is highly amenable to genetic manipulation.

Keywords: Axotomy; Drosophila; Glia; Glial immunity; Neurodegeneration; Phagocytosis; Wallerian degeneration.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Video-Audio Media

MeSH terms

  • Animals
  • Axons / physiology*
  • Axotomy
  • Central Nervous System / pathology
  • DNA, Complementary / genetics
  • Drosophila Proteins / biosynthesis
  • Drosophila Proteins / genetics
  • Drosophila melanogaster / genetics
  • Drosophila melanogaster / immunology*
  • Drosophila melanogaster / physiology
  • Gene Expression Regulation
  • Genes, Reporter
  • Image Processing, Computer-Assisted
  • Immunohistochemistry / instrumentation
  • Immunohistochemistry / methods
  • Intravital Microscopy*
  • Microscopy, Confocal / methods*
  • Nerve Tissue Proteins / biosynthesis
  • Nerve Tissue Proteins / genetics
  • Neuroglia / immunology*
  • Neuroglia / metabolism
  • Phagocytosis
  • Polymerase Chain Reaction / methods
  • Wallerian Degeneration / immunology*
  • Wallerian Degeneration / physiopathology

Substances

  • DNA, Complementary
  • Drosophila Proteins
  • Nerve Tissue Proteins