CaMello-XR enables visualization and optogenetic control of Gq/11 signals and receptor trafficking in GPCR-specific domains

• Published in: Communications biology 2019-02, Vol.2 (1), p.60-60, Article 60
• Main Authors: Eickelbeck, Dennis, Karapinar, Raziye, Jack, Alexander, Suess, Sandra T., Barzan, Ruxandra, Azimi, Zohre, Surdin, Tatjana, Grömmke, Michelle, Mark, Melanie D., Gerwert, Klaus, Jancke, Dirk, Wahle, Petra, Spoida, Katharina, Herlitze, Stefan
• Format: Article
• Language: English
• Subjects: 13/95; 631/378/340; 631/378/87; 631/61/338/469; 631/80/86/1999; 82; 9/74; 96; 96/35; Biology; Biomedical and Life Sciences; C-Terminus; Calcium (intracellular); Calcium signalling; G protein-coupled receptors; Life Sciences; Localization; Melanopsin; Protein transport; Serotonin receptors; Serotonin S2 receptors; Signal transduction; Synaptic plasticity; Visualization
• ISSN: 2399-3642; 2399-3642
• DOI: 10.1038/s42003-019-0292-y

The signal specificity of G protein-coupled receptors (GPCRs) including serotonin receptors (5-HT-R) depends on the trafficking and localization of the GPCR within its subcellular signaling domain. Visualizing traffic-dependent GPCR signals in neurons is difficult, but important to understand the contribution of GPCRs to synaptic plasticity. We engineered CaMello (Ca 2+ -melanopsin-local-sensor) and CaMello-5HT 2A for visualization of traffic-dependent Ca 2+ signals in 5-HT 2A -R domains. These constructs consist of the light-activated G q/11 coupled melanopsin, mCherry and GCaMP6m for visualization of Ca 2+ signals and receptor trafficking, and the 5-HT 2A C-terminus for targeting into 5-HT 2A -R domains. We show that the specific localization of the GPCR to its receptor domain drastically alters the dynamics and localization of the intracellular Ca 2+ signals in different neuronal populations in vitro and in vivo. The CaMello method may be extended to every GPCR coupling to the G q/11 pathway to help unravel new receptor-specific functions in respect to synaptic plasticity and GPCR localization. Dennis Eickelbeck et al. engineered light-activated constructs, CaMello and CaMello-5HT 2A , which are targeted to the 5HT 2A -R domains and enable visualization of calcium signals and receptor trafficking in response to activation. The reported CaMello tool could be applied to other GPCRs coupled to the G q/11 signaling pathways which may shed light on mechanisms of GPCR localization and plasticity.