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Interferon gamma and lipopolysaccharide upregulate TNF-related apoptosis-inducing ligand expression in murine
Sermin Genc1, Sefa Kizildag, Kursad Genc
1Department of Medical Biology and Medical Genetics, School of Medicine, Dokuz Eylul University, Inciralti, 35340, Izmir, Turkey. sermingenc@hotmail.com
Abstract:
In this study, it is reported that neonatal murine microglia and N9 murine microglial cell line express tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL). TRAIL protein and mRNA expression in murine microglia greatly upregulate upon stimulation with interferon gamma (IFNgamma) or lipopolysaccharide (LPS) as revealed by immunoprecipitation-immunoblotting, reverse transcriptase-polymerase chain reaction (RT-PCR) and flow cytometry techniques. IFNgamma and LPS act synergistically to induce TRAIL expression on both translational and transcriptional levels. The upregulated microglial TRAIL in inflammatory conditions may involve in the cytotoxic effect of these cells and play a role in neurodegenerative processes.
Insights
Neonatal microglia and N9 cells express tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL). Interferon gamma (IFNgamma) and lipopolysaccharide (LPS) significantly increase TRAIL expression, potentially contributing to neurodegeneration.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia are the primary immune cells of the central nervous system.
- Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is involved in apoptosis.
- The role of TRAIL in microglia, especially under inflammatory conditions, is not fully understood.
Purpose of the Study:
- To investigate the expression of TRAIL in neonatal murine microglia and the N9 microglial cell line.
- To determine the effect of inflammatory stimuli, such as interferon gamma (IFNgamma) and lipopolysaccharide (LPS), on TRAIL expression.
- To explore the potential role of microglial TRAIL in neurodegenerative processes.
Main Methods:
- Immunoprecipitation-immunoblotting to detect TRAIL protein.
- Reverse transcriptase-polymerase chain reaction (RT-PCR) to quantify TRAIL mRNA.
- Flow cytometry to analyze TRAIL expression on cell surfaces.
Main Results:
- Neonatal murine microglia and N9 cells express TRAIL.
- IFNgamma and LPS stimulation significantly upregulate both TRAIL protein and mRNA levels.
- IFNgamma and LPS act synergistically to induce TRAIL expression at both translational and transcriptional levels.
Conclusions:
- Microglial TRAIL expression is significantly enhanced by inflammatory stimuli like IFNgamma and LPS.
- Upregulated microglial TRAIL in inflammation may contribute to the cytotoxic effects of microglia.
- Microglial TRAIL may play a role in the pathogenesis of neurodegenerative diseases.