Live imaging of microtubule dynamics at excitatory presynaptic boutons in primary hippocampal neurons and acute hippocampal slices

STAR Protoc. 2021 Feb 21;2(1):100342. doi: 10.1016/j.xpro.2021.100342. eCollection 2021 Mar 19.

Abstract

Analyses of microtubule (MT) plus end dynamics at glutamatergic en passant boutons can be carried out in cultured primary neurons isolated from mouse or rat embryos or ex vivo in acute slices isolated from mice that had been electroporated in utero. Here, we describe a protocol for setting up and analyzing live image recordings of primary neurons and acute hippocampal slices expressing tagged versions of the MT plus end binding protein EB3 and the presynaptic vesicle markers vGlut1 or VAMP2. For complete information on the use and execution of this protocol, please refer to Qu et al. (2019).

Keywords: Cell biology; Microscopy; Neuroscience.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Glucose Transporter Type 1 / metabolism
  • Hippocampus / cytology
  • Hippocampus / metabolism*
  • Mice
  • Microtubule-Associated Proteins / metabolism
  • Microtubules / metabolism*
  • Neurons / cytology
  • Neurons / metabolism*
  • Presynaptic Terminals / metabolism*
  • Rats
  • Rats, Sprague-Dawley
  • Vesicle-Associated Membrane Protein 2 / metabolism

Substances

  • EB3 protein, mouse
  • Glucose Transporter Type 1
  • Mapre3 protein, rat
  • Microtubule-Associated Proteins
  • Slc2a1 protein, mouse
  • Slc2a1 protein, rat
  • Vamp2 protein, rat
  • Vesicle-Associated Membrane Protein 2
  • vesicle-associated membrane protein 2, mouse