Substitution of 5-HT1A Receptor Signaling by a Light-activated G Protein-coupled Receptor

Understanding serotonergic (5-HT) signaling is critical for understanding human physiology, behavior, and neuropsychiatric disease. 5-HT mediates its actions via ionotropic and metabotropic 5-HT receptors. The 5-HT(1A) receptor is a metabotropic G protein-coupled receptor linked to the G(i/o) signaling pathway and has been specifically implicated in the pathogenesis of depression and anxiety. To understand and precisely control 5-HT(1A) signaling, we created a light-activated G protein-coupled receptor that targets into 5-HT(1A) receptor domains and substitutes for endogenous 5-HT(1A) receptors. To induce 5-HT(1A)-like targeting, vertebrate rhodopsin was tagged with the C-terminal domain (CT) of 5-HT(1A) (Rh-CT(5-HT1A)). Rh-CT(5-HT1A) activates G protein-coupled inward rectifying K(+) channels in response to light and causes membrane hyperpolarization in hippocampal neurons, similar to the agonist-induced responses of the 5-HT(1A) receptor. The intracellular distribution of Rh-CT(5-HT1A) resembles that of the 5-HT(1A) receptor; Rh-CT(5-HT1A) localizes to somatodendritic sites and is efficiently trafficked to distal dendritic processes. Additionally, neuronal expression of Rh-CT(5-HT1A), but not Rh, decreases 5-HT(1A) agonist sensitivity, suggesting that Rh-CT(5-HT1A) and 5-HT(1A) receptors compete to interact with the same trafficking machinery. Finally, Rh-CT(5-HT1A) is able to rescue 5-HT(1A) signaling of 5-HT(1A) KO mice in cultured neurons and in slices of the dorsal raphe showing that Rh-CT(5-HT1A) is able to functionally compensate for native 5-HT(1A). Thus, as an optogenetic tool, Rh-CT(5-HT1A) has the potential to directly correlate in vivo 5-HT(1A) signaling with 5-HT neuron activity and behavior in both normal animals and animal models of neuropsychiatric disease.