Pharmacological evidence for transactivation within melatonin MT₂ and serotonin 5-HT_2C receptor heteromers in mouse...

Association of G protein-coupled receptors into heterodimeric complexes has been reported for over 50 receptor pairs in vitro but functional in vivo validation remains a challenge. Our recent in vitro studies defined the functional fingerprint of heteromers composed of G_i -coupled melatonin MT₂ receptors and G_q -coupled serotonin 5-HT_2C receptors, in which melatonin transactivates phospholipase C (PLC) through 5-HT_2C . Here, we identified this functional fingerprint in the mouse brain. G_q protein activation was probed by [³⁵ S]GTPγS incorporation followed by G_q immunoprecipitation, and PLC activation by determining the inositol phosphate levels in brain lysates of animals previously treated with melatonin. Melatonin concentration-dependently activated G_q proteins and PLC in the hypothalamus and cerebellum but not in cortex. These effects were inhibited by the 5-HT_2C receptor-specific inverse agonist SB-243213, and were absent in MT₂ and 5-HT_2C knockout mice, fully recapitulating previous in vitro data and indicating the involvement of MT₂ /5-HT_2C heteromers. The antidepressant agomelatine had a similar effect than melatonin when applied alone but blocked the melatonin-promoted G_q activation due to its 5-HT_2C antagonistic component. Collectively, we provide strong functional evidence for the existence of MT₂ /5-HT_2C heteromeric complexes in mouse brain. These heteromers might participate in the in vivo effects of agomelatine.