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Article ; Online: Intravital Imaging of Fluorescent Protein Expression in Mice with a Closed-Skull Traumatic Brain Injury and Cranial Window Using a Two-Photon Microscope.

Zhong, Jianjun / Gunner, Georgia / Henninger, Nils / Schafer, Dorothy P / Bosco, Daryl A

Journal of visualized experiments : JoVE

2023 , Issue 194

Abstract: The goal of this protocol is to demonstrate how to longitudinally visualize the expression and localization of a protein of interest within specific cell types of an animal's brain, upon exposure to exogenous stimuli. Here, the administration of a closed- ...

Abstract	The goal of this protocol is to demonstrate how to longitudinally visualize the expression and localization of a protein of interest within specific cell types of an animal's brain, upon exposure to exogenous stimuli. Here, the administration of a closed-skull traumatic brain injury (TBI) and simultaneous implantation of a cranial window for subsequent longitudinal intravital imaging in mice is shown. Mice are intracranially injected with an adeno-associated virus (AAV) expressing enhanced green fluorescent protein (EGFP) under a neuronal specific promoter. After 2 to 4 weeks, the mice are subjected to a repetitive TBI using a weight drop device over the AAV injection location. Within the same surgical session, the mice are implanted with a metal headpost and then a glass cranial window over the TBI impacting site. The expression and cellular localization of EGFP is examined using a two-photon microscope in the same brain region exposed to trauma over the course of months.
MeSH term(s)	Mice ; Animals ; Skull/surgery ; Brain/diagnostic imaging ; Brain/surgery ; Head ; Brain Injuries, Traumatic/diagnostic imaging ; Coloring Agents ; Intravital Microscopy/methods
Chemical Substances	Coloring Agents
Language	English
Publishing date	2023-04-21
Publishing country	United States
Document type	Journal Article ; Video-Audio Media ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2259946-0
ISSN	1940-087X ; 1940-087X
ISSN (online)	1940-087X
ISSN	1940-087X
DOI	10.3791/64701
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Microglia-astrocyte crosstalk regulates synapse remodeling via Wnt signaling.

Faust, Travis E / Lee, Yi-Han / O'Connor, Ciara D / Boyle, Margaret A / Gunner, Georgia / Badimon, Ana / Ayata, Pinar / Schaefer, Anne / Schafer, Dorothy P

bioRxiv : the preprint server for biology

2024

Abstract: Astrocytes and microglia are emerging key regulators of activity-dependent synapse remodeling that engulf and remove synapses in response to changes in neural activity. Yet, the degree to which these cells communicate to coordinate this process remains ... ...

Abstract	Astrocytes and microglia are emerging key regulators of activity-dependent synapse remodeling that engulf and remove synapses in response to changes in neural activity. Yet, the degree to which these cells communicate to coordinate this process remains an open question. Here, we use whisker removal in postnatal mice to induce activity-dependent synapse removal in the barrel cortex. We show that astrocytes do not engulf synapses in this paradigm. Instead, astrocytes reduce their contact with synapses prior to microglia-mediated synapse engulfment. We further show that reduced astrocyte-contact with synapses is dependent on microglial CX3CL1-CX3CR1 signaling and release of Wnts from microglia following whisker removal. These results demonstrate an activity-dependent mechanism by which microglia instruct astrocyte-synapse interactions, which then provides a permissive environment for microglia to remove synapses. We further show that this mechanism is critical to remodel synapses in a changing sensory environment and this signaling is upregulated in several disease contexts.
Language	English
Publishing date	2024-02-09
Publishing country	United States
Document type	Preprint
DOI	10.1101/2024.02.08.579178
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Article ; Online: Diversity and Function of Glial Cell Types in Multiple Sclerosis.

Schirmer, Lucas / Schafer, Dorothy P / Bartels, Theresa / Rowitch, David H / Calabresi, Peter A

Trends in immunology

2021 Volume 42, Issue 3, Page(s) 228–247

Abstract: Glial subtype diversity is an emerging topic in neurobiology and immune-mediated neurological diseases such as multiple sclerosis (MS). We discuss recent conceptual and technological advances that allow a better understanding of the transcriptomic and ... ...

Abstract	Glial subtype diversity is an emerging topic in neurobiology and immune-mediated neurological diseases such as multiple sclerosis (MS). We discuss recent conceptual and technological advances that allow a better understanding of the transcriptomic and functional heterogeneity of oligodendrocytes (OLs), astrocytes, and microglial cells under inflammatory-demyelinating conditions. Recent single cell transcriptomic studies suggest the occurrence of novel homeostatic and reactive glial subtypes and provide insight into the molecular events during disease progression. Multiplexed RNA in situ hybridization has enabled 'mapping back' dysregulated gene expression to glial subtypes within the MS lesion microenvironment. These findings suggest novel homeostatic and reactive glial-cell-type functions both in immune-related processes and neuroprotection relevant to understanding the pathology of MS.
MeSH term(s)	Astrocytes ; Humans ; Microglia ; Multiple Sclerosis ; Neuroglia ; Oligodendroglia
Language	English
Publishing date	2021-02-13
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S. ; Review
ZDB-ID	2036831-8
ISSN	1471-4981 ; 1471-4906
ISSN (online)	1471-4981
ISSN	1471-4906
DOI	10.1016/j.it.2021.01.005
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Article: A 4D transcriptomic map for the evolution of multiple sclerosis-like lesions in the marmoset brain.

Lin, Jing-Ping / Brake, Alexis / Donadieu, Maxime / Lee, Amanda / Kawaguchi, Riki / Sati, Pascal / Geschwind, Daniel H / Jacobson, Steven / Schafer, Dorothy P / Reich, Daniel S

bioRxiv : the preprint server for biology

2023

Abstract: Single-time-point histopathological studies on postmortem multiple sclerosis (MS) tissue fail to capture lesion evolution dynamics, posing challenges for therapy development targeting development and repair of focal inflammatory demyelination. To close ... ...

Abstract	Single-time-point histopathological studies on postmortem multiple sclerosis (MS) tissue fail to capture lesion evolution dynamics, posing challenges for therapy development targeting development and repair of focal inflammatory demyelination. To close this gap, we studied experimental autoimmune encephalitis (EAE) in the common marmoset, the most faithful animal model of these processes. Using MRI-informed RNA profiling, we analyzed ~600,000 single-nucleus and ~55,000 spatial transcriptomes, comparing them against EAE inoculation status, longitudinal radiological signals, and histopathological features. We categorized 5 groups of microenvironments pertinent to neural function, immune and glial responses, tissue destruction and repair, and regulatory network at brain borders. Exploring perilesional microenvironment diversity, we uncovered central roles of EAE-associated astrocytes, oligodendrocyte precursor cells, and ependyma in lesion formation and resolution. We pinpointed imaging and molecular features capturing the pathological trajectory of WM, offering potential for assessing treatment outcomes using marmoset as a platform.
Language	English
Publishing date	2023-09-27
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.09.25.559371
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Article: Distinct Th17 effector cytokines differentially promote microglial and blood-brain barrier inflammatory responses during post-infectious encephalitis.

Wayne, Charlotte R / Bremner, Luca / Faust, Travis E / Durán-Laforet, Violeta / Ampatey, Nicole / Ho, Sarah J / Feinberg, Philip A / Arvanitis, Panos / Ciric, Bogoljub / Ruan, Chunsheng / Elyaman, Wassim / Delaney, Shannon L / Vargas, Wendy S / Swedo, Susan / Menon, Vilas / Schafer, Dorothy P / Cutforth, Tyler / Agalliu, Dritan

bioRxiv : the preprint server for biology

2023

Abstract: ... Group ... ...

Abstract	Group A
Language	English
Publishing date	2023-05-09
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.03.10.532135
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Article ; Online: A comparative analysis of microglial inducible Cre lines.

Faust, Travis E / Feinberg, Philip A / O'Connor, Ciara / Kawaguchi, Riki / Chan, Andrew / Strasburger, Hayley / Frosch, Maximilian / Boyle, Margaret A / Masuda, Takahiro / Amann, Lukas / Knobeloch, Klaus-Peter / Prinz, Marco / Schaefer, Anne / Schafer, Dorothy P

Cell reports

2023 Volume 42, Issue 9, Page(s) 113031

Abstract: Cre/loxP technology has revolutionized genetic studies and allowed for spatial and temporal control of gene expression in specific cell types. Microglial biology has particularly benefited because microglia historically have been difficult to transduce ... ...

Abstract	Cre/loxP technology has revolutionized genetic studies and allowed for spatial and temporal control of gene expression in specific cell types. Microglial biology has particularly benefited because microglia historically have been difficult to transduce with virus or electroporation methods for gene delivery. Here, we investigate five of the most widely available microglial inducible Cre lines. We demonstrate varying degrees of recombination efficiency, cell-type specificity, and spontaneous recombination, depending on the Cre line and inter-loxP distance. We also establish best practice guidelines and protocols to measure recombination efficiency, particularly in microglia. There is increasing evidence that microglia are key regulators of neural circuits and major drivers of a broad range of neurological diseases. Reliable manipulation of their function in vivo is of utmost importance. Identifying caveats and benefits of all tools and implementing the most rigorous protocols are crucial to the growth of the field and the development of microglia-based therapeutics.
MeSH term(s)	Animals ; Mice ; Microglia/metabolism ; Integrases/metabolism ; Gene Transfer Techniques ; Mice, Transgenic
Chemical Substances	Cre recombinase (EC 2.7.7.-) ; Integrases (EC 2.7.7.-)
Language	English
Publishing date	2023-08-26
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
ZDB-ID	2649101-1
ISSN	2211-1247 ; 2211-1247
ISSN (online)	2211-1247
ISSN	2211-1247
DOI	10.1016/j.celrep.2023.113031
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Article: A comparative analysis of microglial inducible Cre lines.

Faust, Travis E / Feinberg, Philip A / O'Connor, Ciara / Kawaguchi, Riki / Chan, Andrew / Strasburger, Haley / Masuda, Takahiro / Amann, Lukas / Knobeloch, Klaus-Peter / Prinz, Marco / Schaefer, Anne / Schafer, Dorothy P

bioRxiv : the preprint server for biology

2023

Abstract: Cre/LoxP technology has revolutionized genetic studies and allowed for spatial and temporal control of gene expression in specific cell types. The field of microglial biology has particularly benefited from this technology as microglia have historically ... ...

Abstract	Cre/LoxP technology has revolutionized genetic studies and allowed for spatial and temporal control of gene expression in specific cell types. The field of microglial biology has particularly benefited from this technology as microglia have historically been difficult to transduce with virus or electroporation methods for gene delivery. Here, we interrogate four of the most widely available microglial inducible Cre lines. We demonstrate varying degrees of recombination efficiency and spontaneous recombination, depending on the Cre line and loxP distance. We also establish best practice guidelines and protocols to measure recombination efficiency in microglia, which could be extended to other cell types. There is increasing evidence that microglia are key regulators of neural circuit structure and function. Microglia are also major drivers of a broad range of neurological diseases. Thus, reliable manipulation of their function in vivo is of utmost importance. Identifying caveats and benefits of all tools and implementing the most rigorous protocols are crucial to the growth of the field of microglial biology and the development of microglia-based therapeutics.
Language	English
Publishing date	2023-01-09
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.01.09.523268
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Article ; Online: Expression of ALS-PFN1 impairs vesicular degradation in iPSC-derived microglia.

Funes, Salome / Jung, Jonathan / Gadd, Del Hayden / Mosqueda, Michelle / Zhong, Jianjun / Shankaracharya / Unger, Matthew / Stallworth, Karly / Cameron, Debra / Rotunno, Melissa S / Dawes, Pepper / Fowler-Magaw, Megan / Keagle, Pamela J / McDonough, Justin A / Boopathy, Sivakumar / Sena-Esteves, Miguel / Nickerson, Jeffrey A / Lutz, Cathleen / Skarnes, William C /

Lim, Elaine T / Schafer, Dorothy P / Massi, Francesca / Landers, John E / Bosco, Daryl A

Nature communications

2024 Volume 15, Issue 1, Page(s) 2497

Abstract: Microglia play a pivotal role in neurodegenerative disease pathogenesis, but the mechanisms underlying microglia dysfunction and toxicity remain to be elucidated. To investigate the effect of neurodegenerative disease-linked genes on the intrinsic ... ...

Abstract	Microglia play a pivotal role in neurodegenerative disease pathogenesis, but the mechanisms underlying microglia dysfunction and toxicity remain to be elucidated. To investigate the effect of neurodegenerative disease-linked genes on the intrinsic properties of microglia, we studied microglia-like cells derived from human induced pluripotent stem cells (iPSCs), termed iMGs, harboring mutations in profilin-1 (PFN1) that are causative for amyotrophic lateral sclerosis (ALS). ALS-PFN1 iMGs exhibited evidence of lipid dysmetabolism, autophagy dysregulation and deficient phagocytosis, a canonical microglia function. Mutant PFN1 also displayed enhanced binding affinity for PI3P, a critical signaling molecule involved in autophagic and endocytic processing. Our cumulative data implicate a gain-of-toxic function for mutant PFN1 within the autophagic and endo-lysosomal pathways, as administration of rapamycin rescued phagocytic dysfunction in ALS-PFN1 iMGs. These outcomes demonstrate the utility of iMGs for neurodegenerative disease research and implicate microglial vesicular degradation pathways in the pathogenesis of these disorders.
MeSH term(s)	Humans ; Amyotrophic Lateral Sclerosis/metabolism ; Microglia/metabolism ; Induced Pluripotent Stem Cells/metabolism ; Profilins/metabolism ; Neurodegenerative Diseases ; Mutation
Chemical Substances	Profilins ; PFN1 protein, human
Language	English
Publishing date	2024-03-20
Publishing country	England
Document type	Journal Article
ZDB-ID	2553671-0
ISSN	2041-1723 ; 2041-1723
ISSN (online)	2041-1723
ISSN	2041-1723
DOI	10.1038/s41467-024-46695-w
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Article ; Online: Elevated TNF-α Leads to Neural Circuit Instability in the Absence of Interferon Regulatory Factor 8.

Feinberg, Philip A / Becker, Shannon C / Chung, Leeyup / Ferrari, Loris / Stellwagen, David / Anaclet, Christelle / Durán-Laforet, Violeta / Faust, Travis E / Sumbria, Rachita K / Schafer, Dorothy P

The Journal of neuroscience : the official journal of the Society for Neuroscience

2022 Volume 42, Issue 32, Page(s) 6171–6185

Abstract: Interferon regulatory factor 8 (IRF8) is a transcription factor necessary for the maturation of microglia, as well as other peripheral immune cells. It also regulates the transition of microglia and other immune cells to a pro-inflammatory phenotype. ...

Abstract	Interferon regulatory factor 8 (IRF8) is a transcription factor necessary for the maturation of microglia, as well as other peripheral immune cells. It also regulates the transition of microglia and other immune cells to a pro-inflammatory phenotype.
MeSH term(s)	Animals ; Female ; Interferon Regulatory Factors/genetics ; Interferon Regulatory Factors/metabolism ; Male ; Mice ; Multiple Sclerosis/pathology ; Seizures/metabolism ; Seizures/pathology ; Tumor Necrosis Factor-alpha/metabolism
Chemical Substances	Interferon Regulatory Factors ; Tumor Necrosis Factor-alpha ; interferon regulatory factor-8
Language	English
Publishing date	2022-07-05
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
ZDB-ID	604637-x
ISSN	1529-2401 ; 0270-6474
ISSN (online)	1529-2401
ISSN	0270-6474
DOI	10.1523/JNEUROSCI.0601-22.2022
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Zs.A 1668: Show issues

Location:
Je nach Verfügbarkeit (siehe Angabe bei Bestand)
bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
Jg. 1995 - 2021: Lesesall (1.OG)
ab Jg. 2022: Lesesaal (EG)

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Article ; Online: Neuronal signal-regulatory protein alpha drives microglial phagocytosis by limiting microglial interaction with CD47 in the retina.

Jiang, Danye / Burger, Courtney A / Akhanov, Viktor / Liang, Justine H / Mackin, Robert D / Albrecht, Nicholas E / Andrade, Pilar / Schafer, Dorothy P / Samuel, Melanie A

Immunity

2022 Volume 55, Issue 12, Page(s) 2318–2335.e7

Abstract: Microglia utilize their phagocytic activity to prune redundant synapses and refine neural circuits during precise developmental periods. However, the neuronal signals that control this phagocytic clockwork remain largely undefined. Here, we show that ... ...

Abstract	Microglia utilize their phagocytic activity to prune redundant synapses and refine neural circuits during precise developmental periods. However, the neuronal signals that control this phagocytic clockwork remain largely undefined. Here, we show that neuronal signal-regulatory protein alpha (SIRPα) is a permissive cue for microglial phagocytosis in the developing murine retina. Removal of neuronal, but not microglial, SIRPα reduced microglial phagocytosis, increased synpase numbers, and impaired circuit function. Conversely, prolonging neuronal SIRPα expression extended developmental microglial phagocytosis. These outcomes depended on the interaction of presynaptic SIRPα with postsynaptic CD47. Global CD47 deficiency modestly increased microglial phagocytosis, while CD47 overexpression reduced it. This effect was rescued by coexpression of neuronal SIRPα or codeletion of neuronal SIRPα and CD47. These data indicate that neuronal SIRPα regulates microglial phagocytosis by limiting microglial SIRPα access to neuronal CD47. This discovery may aid our understanding of synapse loss in neurological diseases.
MeSH term(s)	Mice ; Animals ; CD47 Antigen/metabolism ; Receptors, Immunologic/metabolism ; Macrophages/metabolism ; Phagocytosis/physiology ; Retina ; Antigens, Differentiation/metabolism
Chemical Substances	CD47 Antigen ; Receptors, Immunologic ; Antigens, Differentiation
Language	English
Publishing date	2022-11-14
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S. ; Research Support, Non-U.S. Gov't
ZDB-ID	1217235-2
ISSN	1097-4180 ; 1074-7613
ISSN (online)	1097-4180
ISSN	1074-7613
DOI	10.1016/j.immuni.2022.10.018
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Zs.MG 69: Show issues

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