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Live Imaging of Autophagosome Biogenesis and Maturation in Primary Neurons

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Imaging and Quantifying Neuronal Autophagy

Part of the book series: Neuromethods ((NM,volume 171))

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Abstract

Live-cell imaging of autophagy in primary neurons has revealed a robust and constitutive pathway for nonselective autophagy in the axon. Autophagosome biogenesis occurs in the distal axon; newly formed autophagosomes engulf cargos including mitochondrial fragments, protein aggregates, and bulk cytoplasm. Once formed, autophagosomes move rapidly and processively along the axon toward the soma, fusing with lysosomes in transit to mature into degradation-competent autolysosomes. Each step of the biogenesis and maturation pathway can be visualized with live imaging of primary neurons in culture; importantly, live imaging in vivo in C. elegans and Drosophila has confirmed observations made in primary neurons. Here, we detail considerations relating to choice of model system, probe, and microscope, and provide advice on methods and specific points to consider.

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Author information

Authors and Affiliations

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

    Andrea K. H. Stavoe & Erika L. F. Holzbaur

  2. Department of Neurobiology and Anatomy, The University of Texas Health Science Center at Houston McGovern Medical School, Houston, TX, USA

    Andrea K. H. Stavoe

Authors
  1. Andrea K. H. Stavoe
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  2. Erika L. F. Holzbaur
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Corresponding author

Correspondence to Erika L. F. Holzbaur .

Editor information

Editors and Affiliations

  1. Department of Physiological Sciences, Stellenbosch University, Stellenbosch, South Africa

    Ben Loos

  2. Healthy Longevity Translational Research Programme, Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Centre for Healthy Longevity National University Health System, Singapore, Singapore

    Esther Wong

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Stavoe, A.K.H., Holzbaur, E.L.F. (2022). Live Imaging of Autophagosome Biogenesis and Maturation in Primary Neurons. In: Loos, B., Wong, E. (eds) Imaging and Quantifying Neuronal Autophagy. Neuromethods, vol 171. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1589-8_3

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  • DOI: https://doi.org/10.1007/978-1-0716-1589-8_3

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1588-1

  • Online ISBN: 978-1-0716-1589-8

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