Novel Microglia Depletion Systems: A Genetic Approach Utilizing Conditional Diphtheria Toxin Receptor Expression and a Pharmacological Model Based on the Blocking of Macrophage Colony-Stimulating Factor 1 Receptor

Methods Mol Biol. 2019:2034:217-230. doi: 10.1007/978-1-4939-9658-2_16.

Abstract

Microglia are the main population of macrophage residing in the central nervous system (CNS). Depletion experiments gave important insights into the physiology and function of microglia in healthy and diseased CNS. Ablation of microglia can be achieved by application of pharmacological or genetic tools. Here, we describe two approaches to ablate microglia: an efficient genetic model that utilizes DTRMG mouse line that has diphtheria toxin receptor (DTR) expression regulated by the promoter activity of the fractalkine receptor (CX3CR1) gene, and a pharmacological model that utilizes the blocking of macrophage colony-stimulating factor 1 receptor (CSF-1R) with a blocking antibody. Both the administration of systemic diphtheria toxin or anti-CSF-1R blocking antibody result in highly efficient and reversible depletion of microglia population in the CNS, which can be easily assessed by flow cytometry.

Keywords: Anti-CSF-1R antibody AFS98; CSF-1R; CX3CR1; Diphtheria toxin receptor; FACS; Microglia depletion; Tamoxifen.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Blocking / pharmacology*
  • CX3C Chemokine Receptor 1 / genetics*
  • Diphtheria Toxin / pharmacology*
  • Gene Expression*
  • Heparin-binding EGF-like Growth Factor* / biosynthesis
  • Heparin-binding EGF-like Growth Factor* / genetics
  • Mice
  • Mice, Transgenic
  • Microglia*
  • Promoter Regions, Genetic*
  • Receptor, Macrophage Colony-Stimulating Factor* / antagonists & inhibitors
  • Receptor, Macrophage Colony-Stimulating Factor* / genetics
  • Receptor, Macrophage Colony-Stimulating Factor* / metabolism

Substances

  • Antibodies, Blocking
  • CX3C Chemokine Receptor 1
  • Cx3cr1 protein, mouse
  • Diphtheria Toxin
  • Heparin-binding EGF-like Growth Factor
  • Receptor, Macrophage Colony-Stimulating Factor