Detection of Synaptic Proteins in Microglia by Flow Cytometry
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 s...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2020-09-01
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| Series: | Frontiers in Molecular Neuroscience |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fnmol.2020.00149/full |
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| author | Simone Brioschi Simone Brioschi Paolo d’Errico Lukas S. Amann Lukas S. Amann Hana Janova Sonja M. Wojcik Melanie Meyer-Luehmann Lawrence Rajendran Peter Wieghofer Rosa C. Paolicelli Rosa C. Paolicelli Knut Biber |
| author_facet | Simone Brioschi Simone Brioschi Paolo d’Errico Lukas S. Amann Lukas S. Amann Hana Janova Sonja M. Wojcik Melanie Meyer-Luehmann Lawrence Rajendran Peter Wieghofer Rosa C. Paolicelli Rosa C. Paolicelli Knut Biber |
| author_sort | Simone Brioschi |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-17T13:17:39Z |
| format | Article |
| id | doaj.art-4212dc9e7c4743df81cd46c466e44482 |
| institution | Directory Open Access Journal |
| issn | 1662-5099 |
| language | English |
| last_indexed | 2024-12-17T13:17:39Z |
| publishDate | 2020-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Molecular Neuroscience |
| spelling | doaj.art-4212dc9e7c4743df81cd46c466e444822022-12-21T21:46:57ZengFrontiers Media S.A.Frontiers in Molecular Neuroscience1662-50992020-09-011310.3389/fnmol.2020.00149561157Detection of Synaptic Proteins in Microglia by Flow CytometrySimone Brioschi0Simone Brioschi1Paolo d’Errico2Lukas S. Amann3Lukas S. Amann4Hana Janova5Sonja M. Wojcik6Melanie Meyer-Luehmann7Lawrence Rajendran8Peter Wieghofer9Rosa C. Paolicelli10Rosa C. Paolicelli11Knut Biber12Faculty of Biology, University of Freiburg, Freiburg, GermanyDepartment of Psychiatry, University of Freiburg Medical Center, Freiburg, GermanyDepartment of Neurology, University of Freiburg Medical Center, Freiburg, GermanyFaculty of Biology, University of Freiburg, Freiburg, GermanyInstitute of Neuropathology, University of Freiburg Medical Center, Freiburg, GermanyDepartment of Clinical Neuroscience, Max Planck Institute of Experimental Medicine, Göttingen, GermanyDepartment of Molecular Neurobiology, Max Planck Institute of Experimental Medicine, Göttingen, GermanyDepartment of Neurology, University of Freiburg Medical Center, Freiburg, GermanyInstitute for Regenerative Medicine, University of Zürich, Zürich, SwitzerlandInstitute of Anatomy, Leipzig University, Leipzig, GermanyInstitute for Regenerative Medicine, University of Zürich, Zürich, SwitzerlandDepartment of Biomedical Sciences, University of Lausanne, Lausanne, SwitzerlandFaculty of Biology, University of Freiburg, Freiburg, GermanyA 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.https://www.frontiersin.org/article/10.3389/fnmol.2020.00149/fullmicrogliasynaptic pruningVGLUT15xFAD modelTDP-43 conditional knock-out |
| spellingShingle | Simone Brioschi Simone Brioschi Paolo d’Errico Lukas S. Amann Lukas S. Amann Hana Janova Sonja M. Wojcik Melanie Meyer-Luehmann Lawrence Rajendran Peter Wieghofer Rosa C. Paolicelli Rosa C. Paolicelli Knut Biber Detection of Synaptic Proteins in Microglia by Flow Cytometry Frontiers in Molecular Neuroscience microglia synaptic pruning VGLUT1 5xFAD model TDP-43 conditional knock-out |
| title | Detection of Synaptic Proteins in Microglia by Flow Cytometry |
| title_full | Detection of Synaptic Proteins in Microglia by Flow Cytometry |
| title_fullStr | Detection of Synaptic Proteins in Microglia by Flow Cytometry |
| title_full_unstemmed | Detection of Synaptic Proteins in Microglia by Flow Cytometry |
| title_short | Detection of Synaptic Proteins in Microglia by Flow Cytometry |
| title_sort | detection of synaptic proteins in microglia by flow cytometry |
| topic | microglia synaptic pruning VGLUT1 5xFAD model TDP-43 conditional knock-out |
| url | https://www.frontiersin.org/article/10.3389/fnmol.2020.00149/full |
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