title: Imaging of motor-dependent transport in neuronal and nonneuronal cells at high spatial and temporal resolution.
abstract: A wide range of subcellular organelles, pathogens, and macromolecular complexes are actively transported within neuronal and nonneuronal cells by microtubule motors. Transport speeds range up to 2-3 μm/s, which requires millisecond- and nanometer-scale resolution for proper imaging and analysis. Dissecting the contributions of multiple motor types has been challenging because of their functional interdependence and the complexity of individual motor behavior. In this chapter, we describe several methods for motor inhibition coupled with high-resolution particle tracking of vesicular and virus cargoes to provide a detailed and quantitative understanding of motor behavior and regulation. We discuss long-term inhibition, as well as short-term inhibition methods when needed to minimize complications from motor protein interactions.
authors.affiliation: Department of Pathology and Cell Biology, Columbia University, New York, NY, USA.
authors.forename: Julie
authors.initials: J
authors.lastname: Yi
authors.affiliation: Department of Pathology and Cell Biology, Columbia University, New York, NY, USA.
authors.forename: Noopur V
authors.initials: NV
authors.lastname: Khobrekar
authors.affiliation: Department of Pathology and Cell Biology, Columbia University, New York, NY, USA.
authors.forename: Tiago J
authors.initials: TJ
authors.lastname: Dantas
authors.affiliation: Department of Pathology and Cell Biology, Columbia University, New York, NY, USA.
authors.forename: Jie
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Department of Pathology and Cell Biology, Columbia University, New York, NY, USA.
authors.forename: Richard B
authors.initials: RB
authors.lastname: Vallee
mesh_terms: D000818:Animals; D001370:Axonal Transport / Q000502:physiology*; D019556:COS Cells; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D056445:Cytoplasmic Dyneins / Q000378:metabolism; D011992:Endosomes / Q000378:metabolism; D006367:HeLa Cells; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D016547:Kinesins / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism; D015388:Organelles / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D014780:Viruses / Q000378:metabolism
chemical_list: D034741:RNA, Small Interfering; D056445:Cytoplasmic Dyneins; D016547:Kinesins
keywords: Adenovirus; Dynein; Live imaging; Lysosomes; Motility inhibition; Motor proteins; Neurons; Particle tracking; Vesicular transport