title: Imaging organelle transport in primary hippocampal neurons treated with amyloid-β oligomers.
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.
authors.affiliation: Department of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, BC, Canada.
authors.forename: Kathlyn J
authors.initials: KJ
authors.lastname: Gan
authors.affiliation: Department of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, BC, Canada; Department of Biological Sciences, Simon Fraser University, Burnaby, BC, Canada; Brain Research Centre, University of British Columbia, Vancouver, BC, Canada.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Silverman
mesh_terms: D000544:Alzheimer Disease / Q000473:pathology; D016229:Amyloid beta-Peptides / Q000378:metabolism*; D000818:Animals; D001370:Axonal Transport / Q000502:physiology*; D001369:Axons / Q000378:metabolism*; D019208:Brain-Derived Neurotrophic Factor / Q000378:metabolism*; D017173:Caenorhabditis elegans / Q000166:cytology; D002470:Cell Survival; D002478:Cells, Cultured; D056445:Cytoplasmic Dyneins / Q000378:metabolism; D004331:Drosophila melanogaster / Q000166:cytology; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism*; D016547:Kinesins / Q000378:metabolism; D007734:Kymography / Q000379:methods; D008164:Luminescent Proteins; D051379:Mice; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism; D061251:Primary Cell Culture; D014162:Transfection; D016875:tau Proteins / Q000378:metabolism; D000097573:Red Fluorescent Protein
chemical_list: D016229:Amyloid beta-Peptides; D019208:Brain-Derived Neurotrophic Factor; D056445:Cytoplasmic Dyneins; D016547:Kinesins; D008164:Luminescent Proteins; D016875:tau Proteins; D000097573:Red Fluorescent Protein; C409504:blue fluorescent protein, Aequorea victoria
keywords: Alzheimer's disease; Amyloid-β oligomers; Axonal transport; Cellular toxicity; Fluorescence microscopy; Lipid-based transfection; Live cell imaging; Primary neuronal culture