RRC ID 54415
Author Dong A, Kulkarni VV, Maday S.
Title Methods for Imaging Autophagosome Dynamics in Primary Neurons.
Journal Methods Mol Biol
Abstract Autophagy is an essential degradative pathway that maintains neuronal homeostasis and prevents axon degeneration. However, the mechanisms of autophagy in neurons are only beginning to be understood. To address this fundamental gap in knowledge, we have established several key methodologies for live-cell imaging and quantitative analysis of autophagy in primary hippocampal neurons. Using these methods, we have defined compartment-specific dynamics of autophagy in real-time under basal versus stress conditions. For example, we have characterized autophagosome biogenesis in the distal axon and subsequent retrograde transport to the soma for degradation. Autophagosomes are also generated locally within the soma. In contrast to the axon, the majority of autophagosomes in dendrites are stationary, while some exhibit bidirectional movement. These studies establish an initial road map for autophagosome dynamics in each compartment of the neuron and set the stage for a more detailed understanding of neuronal autophagy in stress and disease.
Volume 1880
Pages 243-256
Published 2019-1-1
DOI 10.1007/978-1-4939-8873-0_16
PMID 30610702
MeSH Animals Autophagosomes / ultrastructure* Autophagy* Axons / ultrastructure Cell Culture Techniques / methods Cells, Cultured Dendrites / ultrastructure Hippocampus / cytology Mice Mice, Transgenic Microscopy, Confocal / methods Microscopy, Fluorescence / methods* Neuroglia / cytology Neurons / cytology* Neurons / ultrastructure Optical Imaging / methods*
Mice RBRC00806