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Endosomal Transport to Lysosomes and the Trans-Golgi Network in Neurons and Other Cells: Visualizing Maturational Flux

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2557))

Abstract

High-level microscopy enables the comprehensive study of dynamic intracellular processes. Here we describe a toolkit of combinatorial approaches for fixed cell imaging and live cell imaging to investigate the interactions along the trans-Golgi network (TGN)–endosome–lysosome transport axis, which underlie the maturation of endosomal compartments and degradative flux. For fixed cell approaches, we specifically highlight how choices of permeabilization conditions, antibody selection, and antibody multiplexing affect interpretation of results. For live cell approaches, we emphasize the use of sensors that read out pH and degradative capacity in combination with endosomal identity for elucidating dynamic compartment changes.

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Authors and Affiliations

  1. Department of Cell Biology, University of Virginia, Charlottesville, VA, USA

    Chan Choo Yap & Bettina Winckler

Authors
  1. Ryan J. Mulligan
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  2. Chan Choo Yap
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  3. Bettina Winckler
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Corresponding authors

Correspondence to Chan Choo Yap or Bettina Winckler .

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Editors and Affiliations

  1. Department of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI, USA

    Yanzhuang Wang

  2. Department of Physiology and Cell Biology, University of Arkansas for Medical Sciences, Little Rock, AR, USA

    Vladimir V Lupashin

  3. Department of Biological Sciences, Vanderbilt University, Nashville, TN, USA

    Todd R. Graham

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Mulligan, R.J., Yap, C.C., Winckler, B. (2023). Endosomal Transport to Lysosomes and the Trans-Golgi Network in Neurons and Other Cells: Visualizing Maturational Flux. In: Wang, Y., Lupashin, V.V., Graham, T.R. (eds) Golgi. Methods in Molecular Biology, vol 2557. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2639-9_36

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  • DOI: https://doi.org/10.1007/978-1-0716-2639-9_36

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