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Visualizing Vesicle-Bound Kinesins in Cultured Hippocampal Neurons

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Axonal Transport

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2431))

Abstract

Eukaryotic cells use microtubule-based vesicle transport to exchange molecules between compartments. Kinesin family members mediate all microtubule plus end-directed vesicle transport. Of the 45 kinesins expressed in humans, some 20 mediate microtubule plus-end directed vesicle transport. Here we describe a technique to visualize vesicle-bound kinesins in cultured hippocampal neurons. The method involves the expression of the vesicle-binding tail domain while minimizing the cytoplasmic pool. Using this approach drastically improves vesicle labeling compared to full-length kinesins. This tool is useful for systematically comparing the localization of different kinesins in the same cell type and for identifying cargo proteins that reside in vesicles moved by a specific kinesin family member. While we describe the assay in cultured hippocampal neurons, we expect it to be easily transferable to other eukaryotic cell types.

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Acknowledgments

We thank members of the Bentley lab for their feedback on the manuscript. This work was supported by NIH grant R01MH066179 and an NIH-funded predoctoral fellowship to A.G. (T32GM067545).

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Author notes

  1. Andrew Montgomery and Alex Garbouchian contributed equally to this work.

Authors and Affiliations

  1. Department of Biological Sciences and the Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, USA

    Andrew Montgomery, Alex Garbouchian & Marvin Bentley

Authors
  1. Andrew Montgomery
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  2. Alex Garbouchian
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  3. Marvin Bentley
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Corresponding author

Correspondence to Marvin Bentley .

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Editors and Affiliations

  1. Department of Basic and Clinical Neurosciences, Maurice Wohl Clinical Neuroscience Institute, Institute of Psychiatry, Psychology and Neuroscience, King’s College London, London, UK

    Alessio Vagnoni

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Montgomery, A., Garbouchian, A., Bentley, M. (2022). Visualizing Vesicle-Bound Kinesins in Cultured Hippocampal Neurons. In: Vagnoni, A. (eds) Axonal Transport. Methods in Molecular Biology, vol 2431. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1990-2_12

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  • DOI: https://doi.org/10.1007/978-1-0716-1990-2_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1989-6

  • Online ISBN: 978-1-0716-1990-2

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