Imaging organelle transport in primary hippocampal neurons treated with amyloid-β oligomers

Methods Cell Biol. 2016;131:425-51. doi: 10.1016/bs.mcb.2015.06.012. Epub 2015 Sep 2.

Abstract

We describe a strategy for fluorescent imaging of organelle transport in primary hippocampal neurons treated with amyloid-β (Aβ) peptides that cause Alzheimer's disease (AD). This method enables careful, rigorous analyses of axonal transport defects, which are implicated in AD and other neurodegenerative diseases. Moreover, we present and emphasize guidelines for investigating Aβ-induced mechanisms of axonal transport disruption in the absence of nonspecific, irreversible cellular toxicity. This approach should be accessible to most laboratories equipped with cell culture facilities and a standard fluorescent microscope and may be adapted to other cell types.

Keywords: Alzheimer's disease; Amyloid-β oligomers; Axonal transport; Cellular toxicity; Fluorescence microscopy; Lipid-based transfection; Live cell imaging; Primary neuronal culture.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alzheimer Disease / pathology
  • Amyloid beta-Peptides / metabolism*
  • Animals
  • Axonal Transport / physiology*
  • Axons / metabolism*
  • Brain-Derived Neurotrophic Factor / metabolism*
  • Caenorhabditis elegans / cytology
  • Cell Survival
  • Cells, Cultured
  • Cytoplasmic Dyneins / metabolism
  • Drosophila melanogaster / cytology
  • Hippocampus / cytology
  • Hippocampus / metabolism*
  • Kinesin / metabolism
  • Kymography / methods
  • Luminescent Proteins
  • Mice
  • Microscopy, Confocal
  • Microscopy, Fluorescence
  • Microtubules / metabolism
  • Mitochondria / metabolism
  • Primary Cell Culture
  • Transfection
  • tau Proteins / metabolism

Substances

  • Amyloid beta-Peptides
  • Brain-Derived Neurotrophic Factor
  • Luminescent Proteins
  • blue fluorescent protein, Aequorea victoria
  • red fluorescent protein
  • tau Proteins
  • Cytoplasmic Dyneins
  • Kinesin