Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

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

Chunsheng Ruan, Linlin Sun, Alexandra Kroshilina, Lien Beckers, Philip L. De Jager, Elizabeth M. Bradshaw, Samuel Hasson, Guang Yang, Wassim Elyaman
doi: https://doi.org/10.1101/665893
Chunsheng Ruan
aCenter for Translational and Computational Neuroimmunology, Department of Neurology, Columbia University Medical Center, New York, NY 10032, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Linlin Sun
bDepartment of Anesthesiology, Columbia University Medical Center, New York, NY 10032, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Alexandra Kroshilina
aCenter for Translational and Computational Neuroimmunology, Department of Neurology, Columbia University Medical Center, New York, NY 10032, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Lien Beckers
aCenter for Translational and Computational Neuroimmunology, Department of Neurology, Columbia University Medical Center, New York, NY 10032, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Philip L. De Jager
aCenter for Translational and Computational Neuroimmunology, Department of Neurology, Columbia University Medical Center, New York, NY 10032, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Elizabeth M. Bradshaw
aCenter for Translational and Computational Neuroimmunology, Department of Neurology, Columbia University Medical Center, New York, NY 10032, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Samuel Hasson
cPfizer Inc., Cambridge, MA 02141, USA
dAmgen, Cambridge, MA 02141, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Guang Yang
bDepartment of Anesthesiology, Columbia University Medical Center, New York, NY 10032, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Wassim Elyaman
aCenter for Translational and Computational Neuroimmunology, Department of Neurology, Columbia University Medical Center, New York, NY 10032, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: we2152@cumc.columbia.edu
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Supplementary material
  • Preview PDF
Loading

Abstract

Microglia are resident immune cells of the central nervous system (CNS). The exact role of microglia in the physiopathology of CNS disorders is not clear due to lack of tools to discriminate between CNS resident and infiltrated innate immune cells. Here, we present a novel reporter mouse model targeting a microglia-specific marker (TMEM119) for studying the function of 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 mRNA level in the CNS of Tmem119tdTomato/+ relative to control Wild-type mice. The cells expressing tdTomato-were recognized by immunofluorescence staining using commercially available anti-TMEM119 antibodies. Using immunofluorescence and flow cytometry techniques, tdTomato+ cells were detected throughout the CNS, but not in peripheral tissues of adult Tmem119tdTomato/+ mice. In addition, aging does not seem to influence TMEM119 expression as tdTomato+ cells were detectable in the CNS of older mice (300 and 540 days old). Further immunofluorescence characterization shows that the tdTomato+ cells were highly colocalized with Iba1+ cells (microglia and macrophages) in the brain, but not with NeuN- (neurons), GFAP- (astrocytes) or Olig2- (oligodendrocytes) labeled cells. Moreover, flow cytometry analysis of brain tissues of adult mice demonstrates that the majority of microglial CD45lowCD11b+ cells (96.6%) are tdTomato positive. Functionally, using a laser-induced injury model, we measured time-lapse activation of tdTomato-labeled microglia by transcranial two-photon microscopy in live Tmem119tdTomato/+ mice. Taken together, the Tmem119-tdTomato reporter mouse model will serve as a valuable tool to specifically study the role of microglia in health and disease.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
Back to top
PreviousNext
Posted June 10, 2019.
Download PDF

Supplementary Material

Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
A Novel Tmem119-tdTomato Reporter Mouse Model for Studying Microglia in the Central Nervous System
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
A Novel Tmem119-tdTomato Reporter Mouse Model for Studying Microglia in the Central Nervous System
Chunsheng Ruan, Linlin Sun, Alexandra Kroshilina, Lien Beckers, Philip L. De Jager, Elizabeth M. Bradshaw, Samuel Hasson, Guang Yang, Wassim Elyaman
bioRxiv 665893; doi: https://doi.org/10.1101/665893
Digg logo Reddit logo Twitter logo Facebook logo Google logo LinkedIn logo Mendeley logo
Citation Tools
A Novel Tmem119-tdTomato Reporter Mouse Model for Studying Microglia in the Central Nervous System
Chunsheng Ruan, Linlin Sun, Alexandra Kroshilina, Lien Beckers, Philip L. De Jager, Elizabeth M. Bradshaw, Samuel Hasson, Guang Yang, Wassim Elyaman
bioRxiv 665893; doi: https://doi.org/10.1101/665893

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Immunology
Subject Areas
All Articles
  • Animal Behavior and Cognition (3689)
  • Biochemistry (7796)
  • Bioengineering (5675)
  • Bioinformatics (21284)
  • Biophysics (10578)
  • Cancer Biology (8174)
  • Cell Biology (11945)
  • Clinical Trials (138)
  • Developmental Biology (6763)
  • Ecology (10401)
  • Epidemiology (2065)
  • Evolutionary Biology (13866)
  • Genetics (9708)
  • Genomics (13073)
  • Immunology (8146)
  • Microbiology (20014)
  • Molecular Biology (7853)
  • Neuroscience (43056)
  • Paleontology (319)
  • Pathology (1279)
  • Pharmacology and Toxicology (2258)
  • Physiology (3351)
  • Plant Biology (7232)
  • Scientific Communication and Education (1312)
  • Synthetic Biology (2006)
  • Systems Biology (5538)
  • Zoology (1128)