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

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 co...

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Bibliographic Details
Published in:Communications biology 2019, Vol.2, p.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:
Animals
Biosensing Techniques
Calcium - metabolism
Cerebellum - cytology
Cerebellum - metabolism
Cerebral Cortex - cytology
Cerebral Cortex - metabolism
Dependovirus - genetics
Dependovirus - metabolism
Electrodes, Implanted
Gene Expression
Genetic Vectors - chemistry
Genetic Vectors - metabolism
GTP-Binding Protein alpha Subunits, Gq-G11 - genetics
GTP-Binding Protein alpha Subunits, Gq-G11 - metabolism
HEK293 Cells
Humans
Mice
Mice, Inbred C57BL
Neurons - cytology
Neurons - metabolism
Optogenetics - methods
Protein Transport
Rats
Rats, Long-Evans
Receptor, Serotonin, 5-HT2A - genetics
Receptor, Serotonin, 5-HT2A - metabolism
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - metabolism
Rod Opsins - genetics
Rod Opsins - metabolism
Stereotaxic Techniques
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Summary: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 -melanopsin-local-sensor) and CaMello-5HT for visualization of traffic-dependent Ca signals in 5-HT -R domains. These constructs consist of the light-activated G coupled melanopsin, mCherry and GCaMP6m for visualization of Ca signals and receptor trafficking, and the 5-HT C-terminus for targeting into 5-HT -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 signals in different neuronal populations in vitro and in vivo. The CaMello method may be extended to every GPCR coupling to the G pathway to help unravel new receptor-specific functions in respect to synaptic plasticity and GPCR localization.
ISSN:2399-3642