scholarly journals Quantifying the Kinetics of Signaling and Arrestin Recruitment by Nervous System G-Protein Coupled Receptors

2022 ◽  
Vol 15 ◽  
Author(s):  
Sam R. J. Hoare ◽  
Paul H. Tewson ◽  
Shivani Sachdev ◽  
Mark Connor ◽  
Thomas E. Hughes ◽  
...  

Neurons integrate inputs over different time and space scales. Fast excitatory synapses at boutons (ms and μm), and slow modulation over entire dendritic arbors (seconds and mm) are all ultimately combined to produce behavior. Understanding the timing of signaling events mediated by G-protein-coupled receptors is necessary to elucidate the mechanism of action of therapeutics targeting the nervous system. Measuring signaling kinetics in live cells has been transformed by the adoption of fluorescent biosensors and dyes that convert biological signals into optical signals that are conveniently recorded by microscopic imaging or by fluorescence plate readers. Quantifying the timing of signaling has now become routine with the application of equations in familiar curve fitting software to estimate the rates of signaling from the waveform. Here we describe examples of the application of these methods, including (1) Kinetic analysis of opioid signaling dynamics and partial agonism measured using cAMP and arrestin biosensors; (2) Quantifying the signaling activity of illicit synthetic cannabinoid receptor agonists measured using a fluorescent membrane potential dye; (3) Demonstration of multiplicity of arrestin functions from analysis of biosensor waveforms and quantification of the rates of these processes. These examples show how temporal analysis provides additional dimensions to enhance the understanding of GPCR signaling and therapeutic mechanisms in the nervous system.

2018 ◽  
Vol 293 (19) ◽  
pp. 7466-7473 ◽  
Author(s):  
Qingwen Wan ◽  
Najeah Okashah ◽  
Asuka Inoue ◽  
Rony Nehmé ◽  
Byron Carpenter ◽  
...  

2019 ◽  
Vol 14 (6) ◽  
pp. 1141-1149 ◽  
Author(s):  
Robert Serfling ◽  
Lisa Seidel ◽  
Andreas Bock ◽  
Martin J. Lohse ◽  
Paolo Annibale ◽  
...  

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Hui Li ◽  
Jie Yang ◽  
Cuiping Tian ◽  
Min Diao ◽  
Quan Wang ◽  
...  

Abstract G-protein-coupled receptors (GPCRs) play important roles in cellular functions. However, their intracellular organization is largely unknown. Through investigation of the cannabinoid receptor 1 (CB1), we discovered periodically repeating clusters of CB1 hotspots within the axons of neurons. We observed these CB1 hotspots interact with the membrane-associated periodic skeleton (MPS) forming a complex crucial in the regulation of CB1 signaling. Furthermore, we found that CB1 hotspot periodicity increased upon CB1 agonist application, and these activated CB1 displayed less dynamic movement compared to non-activated CB1. Our results suggest that CB1 forms periodic hotspots organized by the MPS as a mechanism to increase signaling efficacy upon activation.


2017 ◽  
Vol 139 (38) ◽  
pp. 13376-13386 ◽  
Author(s):  
Carlo P. Ramil ◽  
Maoqing Dong ◽  
Peng An ◽  
Tracey M. Lewandowski ◽  
Zhipeng Yu ◽  
...  

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