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Updated: Jul 3, 2026
Two-photon Imaging of Microglial Processes' Attraction Toward ATP or Serotonin in Acute Brain Slices
Published on: January 31, 2019
Lysophosphatidic acid-induced membrane ruffling and brain-derived neurotrophic factor gene expression are mediated by
Ryousuke Fujita1, Yan Ma, Hiroshi Ueda
1Division of Molecular Pharmacology and Neuroscience, Nagasaki University Graduate School of Biomedical Sciences, Bunkyo-machi, Nagasaki, Japan.
Abstract:
We examined the effects of lysophosphatidic acid (LPA) on microglia, which may play an important role in the development and maintenance of neuropathic pain. LPA caused membrane ruffling as detected by scanning electron microscopy, and increased the expression of brain-derived neurotrophic factor (BDNF) in a primary culture of rat microglia, which express LPA(3), but not LPA(1) or LPA(2) receptors. These actions were inhibited by a Galpha(q/11)-antisense oligodeoxynucleotide (AS-ODN), U73122, an inhibitor of phospholipase C (PLC), and apyrase, which specifically degrades ATP and ADP. When ATP release was measured using a luciferin-luciferase bioluminescence assay, LPA was shown to increase it in an LPA(3) and PLC inhibitor-reversible manner. However, LPA-induced ATP release was also blocked by the Galpha(q/11) AS-ODN, but not by pertussis toxin. These results suggest that LPA induces the release of ATP from rat primary cultured microglia via the LPA(3) receptor, Galpha(q/11) and PLC, and that the released ATP or ectopically converted ADP may in turn cause membrane ruffling via P2Y(12) receptors and Galpha(i/o) activation, and BDNF expression via activation of P2X(4) receptors.
Insights
Lysophosphatidic acid (LPA) triggers ATP release from microglia, potentially contributing to neuropathic pain. This release involves specific receptors and signaling pathways, impacting microglial function and brain-derived neurotrophic factor (BDNF) expression.
Area of Science:
- Neuroscience
- Cell Biology
- Pain Research
Background:
- Microglia play a crucial role in neuropathic pain.
- Lysophosphatidic acid (LPA) is implicated in pain development and maintenance.
- Understanding microglial responses to LPA is key to developing pain therapies.
Purpose of the Study:
- To investigate the effects of LPA on primary rat microglia.
- To elucidate the signaling pathways involved in LPA-induced microglial responses.
- To determine the role of LPA in ATP release and BDNF expression in microglia.
Main Methods:
- Primary rat microglia cultures were used.
- Scanning electron microscopy detected membrane ruffling.
- LPA receptor expression (LPA3) and signaling pathways (Gαq/11, PLC) were investigated.
- ATP release was quantified using a luciferin-luciferase assay.
- Inhibitors and antisense oligonucleotides were employed to block specific pathways.
Main Results:
- LPA induced membrane ruffling and increased BDNF expression in microglia.
- Microglia expressed LPA3 receptors but not LPA1 or LPA2.
- LPA stimulated ATP release from microglia in an LPA3 and PLC-dependent manner.
- LPA-induced ATP release was mediated by Gαq/11 signaling, not pertussis toxin-sensitive pathways.
- Released ATP/ADP may activate P2Y12 and P2X4 receptors, leading to downstream effects.
Conclusions:
- LPA induces ATP release from microglia via the LPA3 receptor, Gαq/11, and PLC.
- Released ATP/ADP likely mediates LPA's effects on microglial membrane ruffling and BDNF expression through P2 receptors.
- This pathway highlights a novel mechanism in neuropathic pain involving microglial signaling.
