An Engulfment Assay: A Protocol to Assess Interactions Between CNS Phagocytes and Neurons
Open accessAbstract
Phagocytosis is a process in which a cell engulfs material (entire cell, parts of a cell, debris, etc.) in its surrounding extracellular environment and subsequently digests this material, commonly through lysosomal degradation. Microglia are the resident immune cells of the central nervous system (CNS) whose phagocytic function has been described in a broad range of conditions from neurodegenerative disease (e.g., beta-amyloid clearance in Alzheimer's disease) to development of the healthy brain (e.g., synaptic pruning)(1-6). The following protocol is an engulfment assay developed to visualize and quantify microglia-mediated engulfment of presynaptic inputs in the developing mouse retinogeniculate system(7). While this assay was used to assess microglia function in this particular context, a similar approach may be used to assess other phagocytes throughout the brain (e.g., astrocytes) and the rest of the body (e.g., peripheral macrophages) as well as other contexts in which synaptic remodeling occurs (e.g. ,brain injury/disease).
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Analysis of CD68 positive lysosomes and intralysosomal Bassoon within individual microglia was performed based on a published protocol by Schafer et al.
“Analysis of CD68 positive lysosomes as well as intralysosomal Bassooon within individual microglia were performed based on a published protocol by Schafer et al. (57).”
From Control of microglial dynamics by the Arp2/3 complex and the autism- and schizophrenia-associated protein CYFIP1 (2026) · cites this paper as Schafer et al. [57]
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- Authors
- Schafer, D. P., Lehrman, E. K., Heller, C. T., & Stevens, B.
- Affiliations
- Boston Children's Hospital (US)
- Venue
- Journal of Visualized Experiments
- Publisher
- MyJove Corporation
- Published
- 2014-06-08
- Issue
- 88
- ISSN
- 1940-087X (ISSN-L)
- PMC
- PMC4188069
- OpenAlex
- W1977005041
- Cited by
- 108
- References
- 29
- Citations by year
- 2026: 5 · 2025: 14 · 2024: 11 · 2023: 23 · 2022: 15 · 2021: 10 · 2020: 11 · 2019: 8 · 2018: 5 · 2017: 2 · 2016: 3 · 2015: 1
- FWCI
- 1.04
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Journal of Visualized Experiments (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Richard and Susan Smith Family Foundation
- National Institutes of Health (1f32ns066698-01a1)
- National Institute of Neurological Disorders and Stroke (R01-NS-07100801, R01NS071008, F32-NS-066698, R01NS092578)
- Eunice Kennedy Shriver National Institute of Child Health and Human Development (P30HD018655, P30HD-18655)
- Topics
- Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
- Life Sciences › Immunology and Microbiology › Immunology › Phagocytosis and Immune Regulation
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Keywords
- microglia, phagocytosis, microglia-neuron interaction, synaptic pruning, retinal development, astrocytes, macrophages, synaptic remodeling, central nervous system
- MeSH terms
- Animals, Cell Communication, Central Nervous System, Neurons, Phagocytes, Synapses, Microglia, Microscopy, Confocal, Mice
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