Growth cone collapse assay

Methods Mol Biol. 2014:1162:73-83. doi: 10.1007/978-1-4939-0777-9_6.

Abstract

The growth cone collapse assay has proved invaluable in detecting and purifying axonal repellents. Glycoproteins/proteins present in detergent extracts of biological tissues are incorporated into liposomes, added to growth cones in culture and changes in morphology are then assessed. Alternatively purified or recombinant molecules in aqueous solution may be added directly to the cultures. In both cases after a defined period of time (up to 1 h), the cultures are fixed and then assessed by inverted phase contrast microscopy for the percentage of growth cones showing a collapsed profile with loss of flattened morphology, filopodia, and lamellipodia.

MeSH terms

  • Animals
  • Cell Culture Techniques / methods
  • Cells, Cultured
  • Chickens
  • Ganglia, Spinal / cytology*
  • Ganglia, Spinal / drug effects
  • Ganglia, Spinal / metabolism
  • Growth Cones / drug effects
  • Growth Cones / metabolism
  • Growth Cones / ultrastructure*
  • Microscopy, Phase-Contrast / methods*
  • Proteins / administration & dosage
  • Proteins / metabolism
  • Recombinant Proteins / administration & dosage
  • Recombinant Proteins / metabolism
  • Tissue Fixation / methods

Substances

  • Proteins
  • Recombinant Proteins