Live Cell Imaging and Optogenetics-Based Assays for GPCR Activity

Xenia Meshik, Narasimhan Gautam

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages207-221
Number of pages15
DOIs
StatePublished - 2021

Publication series

NameMethods in Molecular Biology
Volume2268
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Cell migration
  • G proteins
  • GPCRs
  • Gβγ translocation
  • Live cell imaging
  • Opsins
  • Optogenetics
  • Second messengers

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