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J Vis Exp DOI:10.3791/51482

An engulfment assay: a protocol to assess interactions between CNS phagocytes and neurons.

Publication TypeJournal Article
Year of Publication2014
AuthorsSchafer, DP, Lehrman, EK, Heller, CT, Stevens, B
JournalJ Vis Exp
Date Published2014 Jun 08
KeywordsAnimals, Cell Communication, Central Nervous System, Mice, Microglia, Microscopy, Confocal, Neurons, Phagocytes, Synapses

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).


Alternate JournalJ Vis Exp
PubMed ID24962472
PubMed Central IDPMC4188069
Grant ListR01 NS084298 / NS / NINDS NIH HHS / United States
F32-NS-066698 / NS / NINDS NIH HHS / United States
F32 NS066698 / NS / NINDS NIH HHS / United States
P30 HD018655 / HD / NICHD NIH HHS / United States
P30-HD-18655 / HD / NICHD NIH HHS / United States
R01-NS-07100801 / NS / NINDS NIH HHS / United States
R01 NS071008 / NS / NINDS NIH HHS / United States
R01 NS092578 / NS / NINDS NIH HHS / United States