Detection of Synaptic Proteins in Microglia by Flow Cytometry
Copyright © 2020 Brioschi, d’Errico, Amann, Janova, Wojcik, Meyer-Luehmann, Rajendran, Wieghofer, Paolicelli and Biber..
A growing body of evidence indicates that microglia actively remove synapses in vivo, thereby playing a key role in synaptic refinement and modulation of brain connectivity. This phenomenon was mainly investigated in immunofluorescence staining and confocal microscopy. However, a quantification of synaptic material in microglia using these techniques is extremely time-consuming and labor-intensive. To address this issue, we aimed to quantify synaptic proteins in microglia using flow cytometry. With this approach, we first showed that microglia from the healthy adult mouse brain contain a detectable level of VGLUT1 protein. Next, we found more than two-fold increased VGLUT1 immunoreactivity in microglia from the developing brain (P15) as compared to adult microglia. These data indicate that microglia-mediated synaptic pruning mostly occurs during the brain developmental period. We then quantified the VGLUT1 staining in microglia in two transgenic models characterized by pathological microglia-mediated synaptic pruning. In the 5xFAD mouse model of Alzheimer's disease (AD) microglia exhibited a significant increase in VGLUT1 immunoreactivity before the onset of amyloid pathology. Moreover, conditional deletion of TDP-43 in microglia, which causes a hyper-phagocytic phenotype associated with synaptic loss, also resulted in increased VGLUT1 immunoreactivity within microglia. This work provides a quantitative assessment of synaptic proteins in microglia, under homeostasis, and in mouse models of disease.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2020 |
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| Erschienen: |
2020 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:13 |
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| Enthalten in: |
Frontiers in molecular neuroscience - 13(2020) vom: 22., Seite 149 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Brioschi, Simone [VerfasserIn] |
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| Links: |
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| Themen: |
5xFAD model |
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| Anmerkungen: |
Date Revised 03.11.2020 published: Electronic-eCollection Citation Status PubMed-not-MEDLINE |
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| doi: |
10.3389/fnmol.2020.00149 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM317012452 |
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| 520 | |a Copyright © 2020 Brioschi, d’Errico, Amann, Janova, Wojcik, Meyer-Luehmann, Rajendran, Wieghofer, Paolicelli and Biber. | ||
| 520 | |a A growing body of evidence indicates that microglia actively remove synapses in vivo, thereby playing a key role in synaptic refinement and modulation of brain connectivity. This phenomenon was mainly investigated in immunofluorescence staining and confocal microscopy. However, a quantification of synaptic material in microglia using these techniques is extremely time-consuming and labor-intensive. To address this issue, we aimed to quantify synaptic proteins in microglia using flow cytometry. With this approach, we first showed that microglia from the healthy adult mouse brain contain a detectable level of VGLUT1 protein. Next, we found more than two-fold increased VGLUT1 immunoreactivity in microglia from the developing brain (P15) as compared to adult microglia. These data indicate that microglia-mediated synaptic pruning mostly occurs during the brain developmental period. We then quantified the VGLUT1 staining in microglia in two transgenic models characterized by pathological microglia-mediated synaptic pruning. In the 5xFAD mouse model of Alzheimer's disease (AD) microglia exhibited a significant increase in VGLUT1 immunoreactivity before the onset of amyloid pathology. Moreover, conditional deletion of TDP-43 in microglia, which causes a hyper-phagocytic phenotype associated with synaptic loss, also resulted in increased VGLUT1 immunoreactivity within microglia. This work provides a quantitative assessment of synaptic proteins in microglia, under homeostasis, and in mouse models of disease | ||
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| 700 | 1 | |a d'Errico, Paolo |e verfasserin |4 aut | |
| 700 | 1 | |a Amann, Lukas S |e verfasserin |4 aut | |
| 700 | 1 | |a Janova, Hana |e verfasserin |4 aut | |
| 700 | 1 | |a Wojcik, Sonja M |e verfasserin |4 aut | |
| 700 | 1 | |a Meyer-Luehmann, Melanie |e verfasserin |4 aut | |
| 700 | 1 | |a Rajendran, Lawrence |e verfasserin |4 aut | |
| 700 | 1 | |a Wieghofer, Peter |e verfasserin |4 aut | |
| 700 | 1 | |a Paolicelli, Rosa C |e verfasserin |4 aut | |
| 700 | 1 | |a Biber, Knut |e verfasserin |4 aut | |
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