@inbook{6a4d017745e745b4b5751bb533e2d165,
title = "Live Cell Imaging and Optogenetics-Based Assays for GPCR Activity",
abstract = "GPCRs are responsible for activation of numerous downstream effectors. Live cell imaging of these effectors therefore provides a real-time readout of GPCR activity and allows for better understanding of temporal dynamics of GPCR-mediated signaling. Opsins, or optically activatable GPCRs, allow for these signaling pathways to be activated in a spatiotemporally precise and reversible manner. Here, we describe optogenetic methods for activating Gi, Gq, and Gs signaling pathways. Additionally, we present assays for detecting activation of these pathways in real time through live cell imaging of Gβγ translocation, PIP3 increase, PIP2 hydrolysis, cAMP production, and cell migration. These assays can be utilized for GPCR-targeted drug development, as well as for studies of a wide range of GPCR-mediated physiological processes.",
keywords = "Cell migration, G proteins, GPCRs, Gβγ translocation, Live cell imaging, Opsins, Optogenetics, Second messengers",
author = "Xenia Meshik and Narasimhan Gautam",
note = "Funding Information: This work was funded by the NIH through NIGMS grants GM069027, GM107370, and GM122577. We thank Akihisa Terakita and Mitsumasa Koyanagi for parapinopsin cDNA and Ignacio Provencio for melanopsin cDNA. Publisher Copyright: {\textcopyright} 2021, Springer Science+Business Media, LLC, part of Springer Nature.",
year = "2021",
doi = "10.1007/978-1-0716-1221-7_14",
language = "English",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "207--221",
booktitle = "Methods in Molecular Biology",
}