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Published on: March 17, 2015
Modulation of TRPM8 alters the phagocytic activity of microglia and induces changes in sub-cellular organelle
Deep Shikha1, Chandan Mahish1, Raima Sing1
1School of Biological Sciences, National Institute of Science Education and Research, An OCC of Homi Bhabha National Institute, Khordha, Jatni, Odisha, 752050, India.
Abstract:
In this work, we investigated the presence and function of TRPM8, a non-selective and cold-sensitive Ca2+-permeable ion channel in the primary microglia cell as well as in microglia cell line BV2. We demonstrate that primary microglia as well as BV2 express TRPM8 endogenously. Both pharmacological activation or inhibition of TRPM8 causes enhanced uptake of bacterial particles at early time points of infection. In BV2, TRPM8 activation and/or LPS-signaling alters its surface expression and cytosolic ROS production. TRPM8 modulation in the absence and presence of LPS causes differential regulation of cytosolic pH and lysosomal pH. Notably, TRPM8 modulation also alters the correlation between lysosomal pH and cytosolic pH depending on TRPM8 modulation and the presence or absence of LPS. Collectively our data suggest that TRPM8 is involved in the regulation of subcellular organelle, i.e. mitochondrial and lysosomal functions. Data also suggest that primarily TRPM8 activation, but often deviation from endogenous TRPM8 function is linked with better innate immune function mediated by microglial cells. We suggest that TRPM8-mediated regulations of sub-cellular organelle functions are more context-dependent manner. Such understanding is relevant in the context of microglial cell functions and innate immunity.
Insights
TRPM8 channels in microglia regulate bacterial particle uptake and organelle function. Modulating TRPM8 enhances innate immunity, suggesting its therapeutic potential for microglial cell functions.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia are key immune cells in the central nervous system.
- TRPM8 (Transient Receptor Potential Melastatin 8) is a cold-sensitive ion channel.
- The role of TRPM8 in microglial function is not well understood.
Purpose of the Study:
- To investigate the expression and function of TRPM8 in primary microglia and BV2 cell lines.
- To determine how TRPM8 modulation affects microglial innate immune responses.
- To explore TRPM8's influence on subcellular organelle function in microglia.
Main Methods:
- Detection of endogenous TRPM8 expression in primary microglia and BV2 cells.
- Pharmacological activation and inhibition of TRPM8.
- Assessment of bacterial particle uptake.
- Analysis of reactive oxygen species (ROS) production.
- Measurement of cytosolic and lysosomal pH.
- Evaluation of mitochondrial and lysosomal function.
Main Results:
- Primary microglia and BV2 cells express TRPM8 endogenously.
- TRPM8 modulation (activation or inhibition) enhances bacterial particle uptake.
- TRPM8 influences surface expression, ROS production, and pH regulation (cytosolic and lysosomal) in BV2 cells.
- TRPM8 affects the correlation between lysosomal and cytosolic pH, particularly with LPS stimulation.
- TRPM8 is implicated in regulating mitochondrial and lysosomal functions.
Conclusions:
- TRPM8 plays a significant role in microglial innate immune function.
- TRPM8 modulation, especially activation, can enhance microglial responses to infection.
- TRPM8's regulation of subcellular organelles is context-dependent and crucial for microglial immunity.
