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Characterization of LAMP1-labeled nondegradative lysosomal and endocytic compartments in neurons

Cheng, Xiu-Tang, Xie, Yu-Xiang, Zhou, Bing, Huang, Ning, Farfel-Becker, Tamar, Sheng, Zu-Hang·2018·Journal of Cell Biology
DOI 10.1083/jcb.201711083PMC PMC6123004
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Abstract

Despite widespread distribution of LAMP1 and the heterogeneous nature of LAMP1-labeled compartments, LAMP1 is routinely used as a lysosomal marker, and LAMP1-positive organelles are often referred to as lysosomes. In this study, we use immunoelectron microscopy and confocal imaging to provide quantitative analysis of LAMP1 distribution in various autophagic and endolysosomal organelles in neurons. Our study demonstrates that a significant portion of LAMP1-labeled organelles do not contain detectable lysosomal hydrolases including cathepsins D and B and glucocerebrosidase. A bovine serum albumin–gold pulse–chase assay followed by ultrastructural analysis suggests a heterogeneity of degradative capacity in LAMP1-labeled endolysosomal organelles. Gradient fractionation displays differential distribution patterns of LAMP1/2 and cathepsins D/B in neurons. We further reveal that LAMP1 intensity in familial amyotrophic lateral sclerosis–linked motor neurons does not necessarily reflect lysosomal deficits in vivo. Our study suggests that labeling a set of lysosomal hydrolases combined with various endolysosomal markers would be more accurate than simply relying on LAMP1/2 staining to assess neuronal lysosome distribution, trafficking, and functionality under physiological and pathological conditions.

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Metadata

Authors
Cheng, X.-T., Xie, Y.-X., Zhou, B., Huang, N., Farfel-Becker, T., & Sheng, Z.-H.
Affiliations
Synaptic Function Section, The Porter Neuroscience Research Center, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD
ORCID
Venue
Journal of Cell Biology
Publisher
Rockefeller University Press
Published
2018-04-25
Volume
217
Issue
9
Pages
3127-3139
ISSN
  • 0021-9525
  • 1540-8140
DOI
10.1083/jcb.201711083
PMC
PMC6123004
Cited by
294
References
5
Language
en
Licence
Funders
  • National Institute of Neurological Disorders and Stroke
  • National Institutes of Health (ZIA NS003029, ZIA NS002946)

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