Quantifying the Kinetics of Signaling and Arrestin Recruitment by Nervous System G-Protein Coupled Receptors
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-couple...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-01-01
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| Series: | Frontiers in Cellular Neuroscience |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fncel.2021.814547/full |
| _version_ | 1819009819099529216 |
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| author | Sam R. J. Hoare Paul H. Tewson Shivani Sachdev Mark Connor Thomas E. Hughes Anne Marie Quinn |
| author_facet | Sam R. J. Hoare Paul H. Tewson Shivani Sachdev Mark Connor Thomas E. Hughes Anne Marie Quinn |
| author_sort | Sam R. J. Hoare |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-21T01:02:25Z |
| format | Article |
| id | doaj.art-52579c6643d047b1bcac5a0da7a49c89 |
| institution | Directory Open Access Journal |
| issn | 1662-5102 |
| language | English |
| last_indexed | 2024-12-21T01:02:25Z |
| publishDate | 2022-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular Neuroscience |
| spelling | doaj.art-52579c6643d047b1bcac5a0da7a49c892022-12-21T19:21:09ZengFrontiers Media S.A.Frontiers in Cellular Neuroscience1662-51022022-01-011510.3389/fncel.2021.814547814547Quantifying the Kinetics of Signaling and Arrestin Recruitment by Nervous System G-Protein Coupled ReceptorsSam R. J. Hoare0Paul H. Tewson1Shivani Sachdev2Mark Connor3Thomas E. Hughes4Anne Marie Quinn5Pharmechanics LLC, Owego, NY, United StatesMontana Molecular, Bozeman, MT, United StatesDepartment of Biomedical Sciences, Macquarie University, Sydney, NSW, AustraliaDepartment of Biomedical Sciences, Macquarie University, Sydney, NSW, AustraliaMontana Molecular, Bozeman, MT, United StatesMontana Molecular, Bozeman, MT, United StatesNeurons 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.https://www.frontiersin.org/articles/10.3389/fncel.2021.814547/fullarrestinbiosensorcannabinoiddynamicsG protein coupled receptor (GPCR)kinetics |
| spellingShingle | Sam R. J. Hoare Paul H. Tewson Shivani Sachdev Mark Connor Thomas E. Hughes Anne Marie Quinn Quantifying the Kinetics of Signaling and Arrestin Recruitment by Nervous System G-Protein Coupled Receptors Frontiers in Cellular Neuroscience arrestin biosensor cannabinoid dynamics G protein coupled receptor (GPCR) kinetics |
| title | Quantifying the Kinetics of Signaling and Arrestin Recruitment by Nervous System G-Protein Coupled Receptors |
| title_full | Quantifying the Kinetics of Signaling and Arrestin Recruitment by Nervous System G-Protein Coupled Receptors |
| title_fullStr | Quantifying the Kinetics of Signaling and Arrestin Recruitment by Nervous System G-Protein Coupled Receptors |
| title_full_unstemmed | Quantifying the Kinetics of Signaling and Arrestin Recruitment by Nervous System G-Protein Coupled Receptors |
| title_short | Quantifying the Kinetics of Signaling and Arrestin Recruitment by Nervous System G-Protein Coupled Receptors |
| title_sort | quantifying the kinetics of signaling and arrestin recruitment by nervous system g protein coupled receptors |
| topic | arrestin biosensor cannabinoid dynamics G protein coupled receptor (GPCR) kinetics |
| url | https://www.frontiersin.org/articles/10.3389/fncel.2021.814547/full |
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