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Updated: Oct 1, 2025
A Novel In Vitro Live-imaging Assay of Astrocyte-mediated Phagocytosis Using pH Indicator-conjugated Synaptosomes
Published on: February 5, 2018
Diminished LC3-Associated Phagocytosis by Huntington's Disease Striatal Astrocytes
Nicole M Wakida1, Alice L Lau2, Jessica Nguyen1
1Biomedical Engineering, Beckman Laser Institute, University of California, Irvine, Irvine, CA, USA.
Background:
In recent years the functions of astrocytes have shifted from conventional supportive roles to also include active roles in altering synapses and engulfment of cellular debris. Recent studies have implicated astrocytes in both protective and pathogenic roles impacting Huntington's disease (HD) progression.
Objective:
The goal of this study is to determine if phagocytosis of cellular debris is compromised in HD striatal astrocytes.
Methods:
Primary adult astrocytes were derived from two HD mouse models; the fast-progressing R6/2 and slower progressing Q175. With the use of laser nanosurgery, a single astrocyte was lysed within an astrocyte network. The phagocytic response of astrocytes was observed with phase contrast and by fluorescence microscopy for GFP-LC3 transiently transfected cells.
Results:
Astrocyte phagocytosis was significantly diminished in primary astrocytes, consistent with the progression of HD in R6/2 and Q175 mouse models. This was defined by the number of astrocytes responding via phagocytosis and by the average number of vesicles formed per cell. GFP-LC3 was found to increasingly localize to phagocytic vesicles over a 20-min imaging period, but not in HD mice, suggesting the involvement of LC3 in astrocyte phagocytosis.
Conclusion:
We demonstrate a progressive decrease in LC3-associated phagocytosis in HD mouse striatal astrocytes.
