New tools for studying microglia in the mouse and human CNS

Proc Natl Acad Sci U S A. 2016 Mar 22;113(12):E1738-46. doi: 10.1073/pnas.1525528113. Epub 2016 Feb 16.

Abstract

The specific function of microglia, the tissue resident macrophages of the brain and spinal cord, has been difficult to ascertain because of a lack of tools to distinguish microglia from other immune cells, thereby limiting specific immunostaining, purification, and manipulation. Because of their unique developmental origins and predicted functions, the distinction of microglia from other myeloid cells is critically important for understanding brain development and disease; better tools would greatly facilitate studies of microglia function in the developing, adult, and injured CNS. Here, we identify transmembrane protein 119 (Tmem119), a cell-surface protein of unknown function, as a highly expressed microglia-specific marker in both mouse and human. We developed monoclonal antibodies to its intracellular and extracellular domains that enable the immunostaining of microglia in histological sections in healthy and diseased brains, as well as isolation of pure nonactivated microglia by FACS. Using our antibodies, we provide, to our knowledge, the first RNAseq profiles of highly pure mouse microglia during development and after an immune challenge. We used these to demonstrate that mouse microglia mature by the second postnatal week and to predict novel microglial functions. Together, we anticipate these resources will be valuable for the future study and understanding of microglia in health and disease.

Keywords: RNAseq; developmental neuroscience; glia; macrophage; microglia.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Animals
  • Antibodies, Monoclonal / immunology
  • Biomarkers
  • Brain / cytology*
  • Brain / embryology
  • Brain / growth & development
  • Cell Division
  • Cell Lineage
  • Child
  • Endotoxemia / pathology
  • Gene Expression Profiling
  • Gene Expression Regulation, Developmental
  • Humans
  • Lipopolysaccharides / toxicity
  • Macrophages / chemistry
  • Membrane Proteins / analysis*
  • Membrane Proteins / deficiency
  • Membrane Proteins / genetics
  • Membrane Proteins / immunology
  • Mice
  • Mice, Knockout
  • Microglia / chemistry*
  • Microglia / physiology
  • Middle Aged
  • Nerve Crush
  • Nerve Tissue Proteins / analysis*
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / immunology
  • Optic Nerve Injuries / pathology
  • Organ Specificity
  • Rabbits
  • Sciatic Nerve / injuries
  • Sciatic Nerve / pathology
  • Sequence Analysis, RNA
  • Temporal Lobe / metabolism
  • Transcriptome

Substances

  • Antibodies, Monoclonal
  • Biomarkers
  • Lipopolysaccharides
  • Membrane Proteins
  • Nerve Tissue Proteins
  • Obif protein, mouse
  • Tmem119 protein, human