Topics

A Novel Tmem119-tdTomato Reporter Mouse Model for Studying Microglia in the Central Nervous System.

08:00 EDT 8th October 2019 | BioPortfolio

Summary of "A Novel Tmem119-tdTomato Reporter Mouse Model for Studying Microglia in the Central Nervous System."

Microglia are resident immune cells of the central nervous system (CNS). The exact role of microglia in CNS disorders is not clear due to lack of tools to discriminate between microglia and infiltrating myeloid cells. Here, we present a novel reporter mouse model targeting a microglia-specific marker, TMEM119, for studying microglia in health and disease. By placing a reporter cassette (GSG-3xFlag-P2A-tdTomato) between the coding sequence of exon 2 and 3'UTR of the Tmem119 gene using CRISPR/Cas9 technology, we generated a Tmem119-tdTomato knock-in mouse strain. Gene expression assay showed no difference of endogenous Tmem119 in the CNS of Tmem119 relative to wild-type mice. The cells expressing tdTomato were recognized by immunofluorescence staining using commercially available anti-TMEM119 antibodies. Additionally, immunofluorescence and flow cytometry techniques revealed that tdTomato cells are detected throughout the CNS, but not in peripheral tissues of Tmem119 mice. Aging does not influence TMEM119 expression as tdTomato cells were detectable in the CNS of older mice (300 and 540 days old). Further immunofluorescence characterization shows that tdTomato cells colocalize with Iba1 cells in the brain, but not with neurons, astrocytes or oligodendrocytes-labeled cells. Moreover, flow cytometry analysis of brain tissues of adult mice demonstrates that the majority of microglia CD45CD11b cells (96.3%) are tdTomato-positive; and a minority of infiltrating CD45CD11b myeloid cells (5.3%) are also tdTomato-positive, which we further characterized and found that tdTomato expression is in part of choroid plexus macrophages but not in meningeal and perivascular macrophages. Functionally, using an acute injury model, we measured time-lapse activation of tdTomato-labeled microglia by transcranial two-photon microscopy in live Tmem119 mice. Taken together, the Tmem119-tdTomato reporter mouse model is a valuable tool to specifically study the role of microglia in health and disease.

Affiliation

Journal Details

This article was published in the following journal.

Name: Brain, behavior, and immunity
ISSN: 1090-2139
Pages:

Links

DeepDyve research library

PubMed Articles [13961 Associated PubMed Articles listed on BioPortfolio]

Targeting microglia using Cx3cr1-cre lines: revisiting the specificity.

Microglia play a pivotal role in maintaining homeostasis of the CNS. There is growing interest in understanding how microglia influence normal brain function and disease progression. Several microglia...

Dermal and muscle fibroblasts and skeletal myofibers survive chikungunya virus infection and harbor persistent RNA.

Chikungunya virus (CHIKV) is an arthritogenic alphavirus that acutely causes fever as well as severe joint and muscle pain. Chronic musculoskeletal pain persists in a substantial fraction of patients ...

Effect of cornel iridoid glycoside on microglia activation through suppression of the JAK/STAT signalling pathway.

The effect of cornel iridoid glycoside (CIG), main component extracted from Cornus officinalis, on microglia activation has not been elucidated so far. We induced a mouse model of multiple sclerosis (...

CD4 T Cells Induce A Subset of MHCII-Expressing Microglia that Attenuates Alzheimer Pathology.

Microglia play a key role in innate immunity in Alzheimer disease (AD), but their role as antigen-presenting cells is as yet unclear. Here we found that amyloid β peptide (Aβ)-specific T helper 1 (...

Generation of Cdx2-mCherry knock-in murine ES cell line to model trophectoderm and intestinal lineage differentiation.

Caudal-type homeobox 2 (.Cdx2) transcription factor is an essential regulator of differentiation to the intestinal epithelium, somatic mesoderm and trophectoderm function in the mouse. However, the re...

Clinical Trials [2834 Associated Clinical Trials listed on BioPortfolio]

Microglia Activation in Schizophrenia

Patients with schizophrenia have volume loss in gray matter. This study is designed to evaluate whether their is microglia activation in schizophrenia using [11C](R)-PK11195 PET.

Peripheral Benzodiazepine Receptors (PBR28) Brain PET Imaging With Lipopolysaccharide Challenge for the Study of Microglia Function in Alzheimer's Disease

To examine the differences in the capacity to activate microglia in patients with Alzheimer's Disease (AD) compared to age-comparable cognitively normal subjects and younger healthy contro...

Role of Chemokine and Chemokine Receptor in Psoriasis

This study aims to elucidate the role of Chemokine and chemokine receptor in the pathogenesis of Psoriasis by using human psoriasis skin xenograft SCID mouse model. The hypothesis is that ...

Endocannabinoid Control of Microglia Activation as a New Therapeutic Target in the Treatment of Schizophrenia

The main objective of this study is to compare microglia activation as measured with PET in combination with the tracer [11C]-R-PK11195 between recent-onset schizophrenia patients who are ...

Subcutaneous Immunotherapy for Mouse in Adults

This is an open label trial of mouse allergenic extract administered by subcutaneous injection in adults with asthma and mouse sensitivity. The study is designed to evaluate: - the saf...

Medical and Biotech [MESH] Definitions

One of the first INBRED MOUSE STRAINS to be sequenced. This strain is commonly used as genetic background for transgenic mouse models. Refractory to many tumors, this strain is also preferred model for studying role of genetic variations in development of diseases.

A mouse substrain that is genetically predisposed to the development of systemic lupus erythematosus-like syndrome, which has been found to be clinically similar to the human disease. It has been determined that this mouse strain carries a mutation in the fas gene. Also, the MRL/lpr is a useful model to study behavioral and cognitive deficits found in autoimmune diseases and the efficacy of immunosuppressive agents.

The third type of glial cell, along with astrocytes and oligodendrocytes (which together form the macroglia). Microglia vary in appearance depending on developmental stage, functional state, and anatomical location; subtype terms include ramified, perivascular, ameboid, resting, and activated. Microglia clearly are capable of phagocytosis and play an important role in a wide spectrum of neuropathologies. They have also been suggested to act in several other roles including in secretion (e.g., of cytokines and neural growth factors), in immunological processing (e.g., antigen presentation), and in central nervous system development and remodeling.

A strain of Rattus norvegicus with elevated blood pressure used as a model for studying hypertension and stroke.

Experimentally induced neoplasms of CONNECTIVE TISSUE in animals to provide a model for studying human SARCOMA.

Quick Search


DeepDyve research library

Relevant Topics

Neurology - Central Nervous System (CNS)
Alzheimer's Disease Anesthesia Anxiety Disorders Autism Bipolar Disorders Dementia Epilepsy Multiple Sclerosis (MS) Neurology Pain Parkinson's Disease Sleep Disorders Neurology is the branch of me...

Bioinformatics
Bioinformatics is the application of computer software and hardware to the management of biological data to create useful information. Computers are used to gather, store, analyze and integrate biological and genetic information which can then be applied...

Gene Expression
The process of gene expression is used by eukaryotes, prokaryotes, and viruses to generate the macromolecular machinery for life. Steps in the gene expression process may be modulated, including the transcription, RNA splicing, translation, and post-tran...


Searches Linking to this Article