GqPCR-mediated signalling in the spotlight: from visualization towards dissection and quantification.

Area of Science:

• Cellular biology
• Molecular pharmacology
• Biophysics

Background:

• G protein-coupled receptors (GPCRs) are vital for human physiology and represent key drug targets.
• Approximately 40% of GPCRs signal through the Gq protein.
• Existing biochemical and crystallographic methods have advanced understanding of GPCRs and their interactions.

Purpose of the Study:

• To review functional imaging tools for visualizing and quantifying GPCR signaling at the single-cell level.
• To highlight advancements in Förster Resonance Energy Transfer (FRET) and Bioluminescence Resonance Energy Transfer (BRET) for signal readout.
• To discuss protein labeling strategies and manipulation tools for studying GPCRs in living systems.

Main Methods:

• Utilizing FRET and BRET for real-time monitoring of GPCR signaling dynamics.
• Employing advanced protein labeling techniques for visualization in live cells.
• Implementing chemically inducible dimerization and optogenetic tools for precise manipulation of signaling pathways.

Main Results:

• Development of sophisticated functional imaging tools for dissecting GPCR signaling.
• Demonstration of FRET and BRET as powerful methods for quantifying signaling events.
• Availability of novel tools for manipulating specific steps in GPCR pathways.

Conclusions:

• Functional imaging technologies provide powerful means to study GPCR signaling in unprecedented detail.
• These tools offer deep insights into Gq-coupled GPCRs (GqPCRs) in living cells and model organisms.
• Advancements in imaging and manipulation tools pave the way for novel therapeutic strategies targeting GPCRs.