FEAST: A flow cytometry-based toolkit for interrogating microglial engulfment of synaptic and myelin proteins

Lasse Dissing-Olesen, Alec J. Walker, Qian Feng, Helena J. Barr, Alicia C. Walker, Lili Xie, Daniel K. Wilton, Indrani Das, Larry I. Benowitz and Beth Stevens ()
Additional contact information
Lasse Dissing-Olesen: F.M. Kirby Neurobiology Center
Alec J. Walker: F.M. Kirby Neurobiology Center
Qian Feng: F.M. Kirby Neurobiology Center
Helena J. Barr: F.M. Kirby Neurobiology Center
Alicia C. Walker: F.M. Kirby Neurobiology Center
Lili Xie: F.M. Kirby Neurobiology Center
Daniel K. Wilton: F.M. Kirby Neurobiology Center
Indrani Das: F.M. Kirby Neurobiology Center
Larry I. Benowitz: F.M. Kirby Neurobiology Center
Beth Stevens: F.M. Kirby Neurobiology Center

Nature Communications, 2023, vol. 14, issue 1, 1-15

Abstract: Abstract Although engulfment is a hallmark of microglia function, fully validated platforms that facilitate high-throughput quantification of this process are lacking. Here, we present FEAST (Flow cytometric Engulfment Assay for Specific Target proteins), which enables interrogation of in vivo engulfment of synaptic material by brain resident macrophages at single-cell resolution. We optimize FEAST for two different analyses: quantification of fluorescent material inside live cells and of engulfed endogenous proteins within fixed cells. To overcome false-positive engulfment signals, we introduce an approach suitable for interrogating engulfment in microglia from perfusion-fixed tissue. As a proof-of-concept for the specificity and versatility of FEAST, we examine the engulfment of synaptic proteins after optic nerve crush and of myelin in two mouse models of demyelination (treatment with cuprizone and injections of lysolecithin). We find that microglia, but not brain-border associated macrophages, engulf in these contexts. Our work underscores how FEAST can be utilized to gain critical insight into functional neuro-immune interactions that shape development, homeostasis, and disease.

Date: 2023
References: View references in EconPapers View complete reference list from CitEc
Citations:

Downloads: (external link)
https://www.nature.com/articles/s41467-023-41448-7 Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:14:y:2023:i:1:d:10.1038_s41467-023-41448-7

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-023-41448-7

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().