Performing Axon Orientation Assays with Secreted Semaphorins and Other Guidance Cues

Methods Mol Biol. 2017;1493:237-246. doi: 10.1007/978-1-4939-6448-2_17.


The guidance of axons within the developing nervous system is orchestrated by a variety of cues that successively and complementary attract or repel axons to achieve a stereotyped wiring of neural circuits. Here we present a version of a method that has been widely used to identify and characterize the effect of guidance cues on the orientation of axons. We describe the coculture, within a three-dimensional environment, of dorsal spinal cord explants together with a cell aggregate secreting a candidate cue and the method to quantify the effect of this cue on axon orientation.

Keywords: Axon guidance; Cell aggregate; Coculture; Plasma clot; Spinal cord explants.

MeSH terms

  • Animals
  • Axons / physiology*
  • Coculture Techniques
  • Female
  • HEK293 Cells
  • Humans
  • Mice
  • Pregnancy
  • Semaphorins / metabolism*
  • Spinal Cord / cytology*
  • Spinal Cord / metabolism


  • Semaphorins