Serotonin receptor oligomerization regulates cAMP-based signaling

Protein-protein interaction is often investigated using quantitative molecular microscopy with Förster resonant energy transfer (FRET). Here, we combined 'linear unmixing FRET' (lux-FRET) with the simultaneous application of a FRET-based biosensor for cAMP to investigate the oligomerization between the 5-HT₇ receptor (5-HT₇R, also known as HTR7) and the 5-HT_1A receptor (5-HT_1AR, also known as HTR1A) and its importance for cAMP signaling. We found that the 5-HT₇R not only stimulates cAMP production, but also forms hetero-oligomers with 5-HT_1AR, which blocks the inhibitory effect of the latter. 5-HT₇R signaling, however, is not affected by this hetero-oligomerization. By modeling the kinetics of intracellular cAMP level changes in relation to the 5-HT₇R:5-HT_1AR stoichiometry, we were able to decipher the complex signaling characteristics of endogenous serotonin receptors in cultured hippocampal neurons. Our findings indicate that serotonergic signaling is not only modulated by the concentration of an individual receptor but also by its specific interaction with other receptors in endogenous systems. We conclude that the regulated ratio of serotonin receptors in immature and mature neurons may be critically involved in both the onset and response to treatments of psychiatric diseases, such as anxiety and depression.