Ligand-induced activation of ERK1/2 signaling by constitutively active G_s-coupled 5-HT receptors

5-HT₄R, 5-HT₆R, and 5-HT_7AR are three constitutively active G_s-coupled 5-HT receptors that have key roles in brain development, learning, memory, cognition, and other physiological processes in the central nervous system. In addition to G_s signaling cascade mediated by these three 5-HT receptors, the ERK1/2 signaling which is dependent on cyclic adenosine monophosphate (cAMP) production and protein kinase A (PKA) activation downstream of G_s signaling has also been widely studied. In this study, we investigated these two signaling pathways originating from the three G_s-coupled 5-HT receptors in AD293 cells. We found that the phosphorylation and activation of ERK1/2 are ligand-induced, in contrast to the constitutively active G_s signaling. This indicates that G_s signaling alone is not sufficient for ERK1/2 activation in these three 5-HT receptors. In addition to G_s, we found that β-arrestin and Fyn are essential for the activation of ERK1/2. Together, these results put forth a novel mechanism for ERK1/2 activation involving the cooperative action of G_s, β-arrestin, and Fyn.