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Updated: Jun 26, 2026
Primary Microglia Isolation from Mixed Glial Cell Cultures of Neonatal Rat Brain Tissue
Published on: August 15, 2012
Genetic inactivation of TRPM4 does not alter the temperature-dependent movement of mouse microglia
Rei Nishimoto1, Yoshikazu Matsuoka2, Makoto Tominaga3
1Department of Anesthesiology and Resuscitology, Okayama University Medical School, Okayama, Japan; Department of Anesthesiology, National Hospital Organization Fukuyama Medical Center, Hiroshima, Japan.
Abstract:
Temperature sensing shapes behavior and cellular functions, yet the molecular basis of thermosensitivity in non-neuronal cells remains poorly defined. Microglia are resident immune cells of the central nervous system that help maintain brain homeostasis via immune surveillance and injury responses. We previously showed that microglial motility is temperature dependent and is largely mediated by the thermosensitive ion channel transient receptor potential vanilloid 4 (TRPV4), a thermosensitive ion channel. However, the contribution of transient receptor potential melastatin 4 (TRPM4) is unclear because suitable Trpm4 mutant mice were not available in our earlier work. Here, we generated functional Trpm4-knockout mice (TRPM4KO) using CRISPR/Cas9 genome editing based on a published strategy. Time-lapse imaging of primary microglia across a range of temperatures revealed that Trpm4 deficiency did not alter temperature-dependent motility in vitro. These results indicate that TRPM4 is dispensable for temperature-dependent microglial motility.
Insights
Transient receptor potential melastatin 4 (TRPM4) does not affect temperature-dependent microglial motility. Studies show TRPM4 is dispensable for this key immune cell function in the central nervous system.
Area of Science:
- Neuroimmunology
- Cellular Physiology
- Ion Channel Function
Background:
- Thermosensitivity influences cellular functions, but its molecular basis in non-neuronal cells like microglia is not well understood.
- Microglia, the brain's resident immune cells, exhibit temperature-dependent motility, previously linked to the TRPV4 ion channel.
- The role of the TRPM4 ion channel in microglial thermosensitivity was previously uninvestigated due to a lack of suitable animal models.
Purpose of the Study:
- To investigate the role of the transient receptor potential melastatin 4 (TRPM4) ion channel in temperature-dependent microglial motility.
- To generate and utilize functional Trpm4-knockout (TRPM4KO) mice to assess TRPM4's contribution.
Main Methods:
- Generation of functional Trpm4-knockout mice using CRISPR/Cas9 genome editing.
- Primary microglia isolation and culture.
- Time-lapse imaging of microglial motility across various temperatures.
Main Results:
- Trpm4 deficiency did not alter the temperature-dependent motility of primary microglia in vitro.
- TRPM4 knockout mice were successfully generated and utilized for these experiments.
Conclusions:
- The transient receptor potential melastatin 4 (TRPM4) ion channel is not essential for temperature-dependent microglial motility.
- TRPV4 remains the primary ion channel mediating temperature-dependent microglial movement, while TRPM4 plays a dispensable role.
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