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Employing Live-Cell Imaging to Study Motor-Mediated Transport

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Dynein

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2623))

Abstract

Microtubule-based transport is a highly regulated process, requiring kinesin and/or dynein motors, a multitude of motor-associated regulatory proteins including activating adaptors and scaffolding proteins, and microtubule tracks that also provide regulatory cues. While in vitro studies are invaluable, fully replicating the physiological conditions under which motility occurs in cells is not yet possible. Here, we describe two methods that can be employed to study motor-based transport and motor regulation in a cellular context. Live-cell imaging of organelle transport in neurons leverages the uniform polarity of microtubules in axons to better understand the factors regulating microtubule-based motility. Peroxisome recruitment assays allow users to examine the net effect of motors and motor-regulatory proteins on organelle distribution. Together, these methods open the door to motility experiments that more fully interrogate the complex cellular environment.

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Acknowledgments

We thank Mariko Tokito, Chantell Evans, and Claire VanDuyne for technical assistance. This research was supported by NSF Graduate Research Fellowship (DGE-1845298) to S.E.C., NIH grant T32 GM008216 to A.R.F., and R35 GM126950 and RM1 GM136511 to E.L.F.H. The authors declare no competing financial interests.

Author information

Authors and Affiliations

  1. Department of Physiology, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, USA

    Sydney E. Cason, Adam R. Fenton & Erika L. F. Holzbaur

  2. Neuroscience Graduate Group, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, USA

    Sydney E. Cason & Erika L. F. Holzbaur

  3. Pennsylvania Muscle Institute, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, USA

    Sydney E. Cason, Adam R. Fenton & Erika L. F. Holzbaur

  4. Cell and Molecular Biology Graduate Group, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, USA

    Adam R. Fenton & Erika L. F. Holzbaur

Authors
  1. Sydney E. Cason
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  2. Adam R. Fenton
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  3. Erika L. F. Holzbaur
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Corresponding author

Correspondence to Erika L. F. Holzbaur .

Editor information

Editors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, CO, USA

    Steven M. Markus

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Cason, S.E., Fenton, A.R., Holzbaur, E.L.F. (2023). Employing Live-Cell Imaging to Study Motor-Mediated Transport. In: Markus, S.M. (eds) Dynein. Methods in Molecular Biology, vol 2623. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2958-1_3

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  • DOI: https://doi.org/10.1007/978-1-0716-2958-1_3

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2957-4

  • Online ISBN: 978-1-0716-2958-1

  • eBook Packages: Springer Protocols

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