Serotonin 5‐HT2A receptor activation induces 2‐arachidonoylglycerol release through a phospholipase c‐dependent mechanism

To date, several studies have demonstrated that phospholipase C‐coupled receptors stimulate the production of endocannabinoids, particularly 2‐arachidonoylglycerol. There is now evidence that endocannabinoids are involved in phospholipase C‐coupled serotonin 5‐HT2A receptor‐mediated behavioral effects in both rats and mice. The main objective of this study was to determine whether activation of the 5‐HT2A receptor leads to the production and release of the endocannabinoid 2‐arachidonoylglycerol. NIH3T3 cells stably expressing the rat 5‐HT2A receptor were first incubated with [3H]‐arachidonic acid for 24 h. Following stimulation with 10 μm serotonin, lipids were extracted from the assay medium, separated by thin layer chromatography, and analyzed by liquid scintillation counting. Our results indicate that 5‐HT2A receptor activation stimulates the formation and release of 2‐arachidonoylglycerol. The 5‐HT2A receptor‐dependent release of 2‐arachidonoylglycerol was partially dependent on phosphatidylinositol‐specific phospholipase C activation. Diacylglycerol produced downstream of 5‐HT2A receptor‐mediated phospholipase D or phosphatidylcholine‐specific phospholipase C activation did not appear to contribute to 2‐arachidonoylglycerol formation in NIH3T3–5HT2A cells. In conclusion, our results support a functional model where neuromodulatory neurotransmitters such as serotonin may act as regulators of endocannabinoid tone at excitatory synapses through the activation of phospholipase C‐coupled G‐protein coupled receptors.

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