Characterization of phenotype markers and neuronotoxic potential of polarised primary microglia in vitro

• DOI: 10.1016/j.bbi.2013.02.005
• Publication Date: 2013-02-27T15:08:13Z
• Authors: Vibol Chhor; Tifenn Le Charpen...; Sophie Lebon; Marie-Virgine Oré; Idoia Lara Celador; Julien Josserand; Vincent Degos; Etienne Jacotot; Henrik Hagberg; Karin Sävman; Carina Mallard; Pierre Gressens; Bobbi Fleiss
• Keywords: Lipopolysaccharides; Male; 0301 basic medicine; Macrophage; M1-M2; Fluorescent Antibody Technique; Gene Expression; MACROPHAGE ACTIVATION; Behavioral Neuroscience; Mice; M1–M2; Neuroinflammation; Drug-screening; TLR4; Cerebral Cortex; Neurons; 0303 health sciences; AMEBOID MICROGLIA; Cell Polarity; Immunohistochemistry; Neuroprotection; Phenotype; Cytokines; Female; Microglia; SPINAL-CORD; HYPOXIA-ISCHEMIA; Chemokines; 570; Cell Survival; Immunology; Primary Cell Culture; TRAUMATIC BRAIN-INJURY; CYTOKINE RESPONSE; 610; Lipopolysaccharide; Real-Time Polymerase Chain Reaction; Article; 03 medical and health sciences; Animals; ISCHEMIC-INJURY; [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]; ALTERNATIVE ACTIVATION; Inflammation; NITRIC-OXIDE; Endocrine and Autonomic Systems; Interleukins; NECROSIS-FACTOR-ALPHA; Toll-Like Receptor 4; Neuronal cell death; RNA

Microglia mediate multiple facets of neuroinflammation, including cytotoxicity, repair, regeneration, and immunosuppression due to their ability acquire diverse activation states, or phenotypes. Modulation microglial phenotype is an appealing neurotherapeutic strategy but a comprehensive study classical more novel phenotypic markers in vitro lacking. The aim this was outline the temporal expression battery from polarised microglia generate tool for screening immunomodulatory potential compounds. We characterised thirty-one macrophage/microglial primary over time (4, 12, 36, 72 h), using RT-qPCR multiplex protein assay. Firstly, we selected Interleukin-4 (IL-4) lipopolysaccharide (LPS) as strongest M1-M2 polarising stimuli, six stimuli tested. At each point, useful identify that were M1 included iNOS, Cox-2 IL-6 loss M2a markers. Markers quantifying M2b-immunomodulatory included, increased IL-1RA SOCS3 M2a-repair arginase-1, M2b discriminatory. Additional regulated at fewer points, are still likely important monitor when assessing therapies. Further, facilitate identification how treatments alter functional affects microglia, soluble products affected type rate neuronal death; M1/2b induced increasing M2a-induced decreasing loss. also assessed any effects prior state, provide way compound may depending on stage injury/insult progression. identified generally reduced switch M2a. Altogether, have profile mechanism outcome can use reference guide first-line therapies search viable neuroprotectants.