1. Academic Validation
  2. Phenotypic Differences in Primary Murine Microglia Treated with NOD1, NOD2, and NOD1/2 Agonists

Phenotypic Differences in Primary Murine Microglia Treated with NOD1, NOD2, and NOD1/2 Agonists

  • J Mol Neurosci. 2020 Apr;70(4):600-609. doi: 10.1007/s12031-019-01466-x.
Susanne Wasmuth 1 Tida Viola Jalilvand 1 2 Björn Laffer 1 3 Martin Busch 1 Dirk Bauer 1 Thomas Langmann 4 Solon Thanos 5 Maren Kasper 1 Arnd Heiligenhaus 6 7
Affiliations

Affiliations

  • 1 Ophtha-Lab, Ophthalmic Center at St.Franziskus Hospital, Münster, Germany.
  • 2 Westphalian Wilhelms University, Münster, Germany.
  • 3 University of Duisburg-Essen, Essen, Germany.
  • 4 Laboratory for Experimental Immunology of the Eye, Department of Ophthalmology, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.
  • 5 Department of Experimental Ophthalmology, Westphalian Wilhelms University of Münster, Münster, Germany.
  • 6 Ophtha-Lab, Ophthalmic Center at St.Franziskus Hospital, Münster, Germany. arnd.heiligenhaus@uveitis-zentrum.de.
  • 7 University of Duisburg-Essen, Essen, Germany. arnd.heiligenhaus@uveitis-zentrum.de.
Abstract

The purpose of the study was studying the influence of different NOD agonists on the morphological phenotype of primary murine microglia and to examine their influence on characteristic cytokines. Primary CD11b-positive cells were isolated from the brain of neonatal mice. The microglial phenotype of the cells was examined by ionized calcium-binding adapter molecule (Iba)1 staining. After14 days in culture, these cells were stimulated by iE-DAP, L18-MDP, or M-TriDAP as NOD1, NOD2, and NOD1/2 agonists, respectively. The cellular morphology was recorded and compared to the phenotype of cells cultured in medium alone or after LPS stimulation. The cells developed a specific phenotype only after treatment with the NOD2 agonist L18-MDP. These cells were characterized by straight extensions carrying tiny spikes and had a high ramification index. This was in sharp contrast to all other treatments, which always resulted in an amoeboid phenotype typically shown by activated microglia in vivo and by cultured microglia in vitro. The staining intensity of IL-6 and TNF-α did not reveal any clear difference independent of the NOD agonist treatment. In contrast, an increased staining intensity was observed for IL-10 after L18-MDP treatment. The NOD2 agonist L18-MDP induced a morphologically distinct phenotype characterized by microspike-decorated dendritiform extensions and a high degree of ramification in primary murine microglia. Increased ramification index and elevated staining intensity of anti-inflammatory IL-10 as hallmarks suggest that a M2-like phenotype of microglia was induced.

Keywords

In vitro study; Microglia; NOD1/2 agonists; Phenotype.

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