Super-Resolution Imaging of G Protein-Coupled Receptors Using Ground State Depletion Microscopy

2019 ◽  
pp. 323-336
Author(s):  
Fabiana A. Caetano Crowley ◽  
Bryan Heit ◽  
Stephen S. G. Ferguson
Keyword(s):  
G Protein ◽  
Ground State ◽  
Super Resolution ◽  
G Protein Coupled Receptors ◽  
Resolution Imaging ◽  
G Protein Coupled

scholarly journals Actin filaments partition primary cilia membranes into distinct fluid corrals

2018 ◽  
Vol 217 (8) ◽  
pp. 2831-2849 ◽  
Author(s):  
Sungsu Lee ◽  
Han Yen Tan ◽  
Ivayla I. Geneva ◽  
Aleksandr Kruglov ◽  
Peter D. Calvert
Keyword(s):  
Diffusion Coefficient ◽  
G Protein ◽  
Primary Cilia ◽  
Super Resolution ◽  
G Protein Coupled Receptors ◽  
Membrane Diffusion ◽  
Filamentous Actin ◽  
Spatiotemporal Resolution ◽  
Fast Tracking ◽  
G Protein Coupled

Physical properties of primary cilia membranes in living cells were examined using two independent, high-spatiotemporal-resolution approaches: fast tracking of single quantum dot–labeled G protein–coupled receptors and a novel two-photon super-resolution fluorescence recovery after photobleaching of protein ensemble. Both approaches demonstrated the cilium membrane to be partitioned into corralled domains spanning 274 ± 20 nm, within which the receptors are transiently confined for 0.71 ± 0.09 s. The mean membrane diffusion coefficient within the corrals, Dm1 = 2.9 ± 0.41 µm2/s, showed that the ciliary membranes were among the most fluid encountered. At longer times, the apparent membrane diffusion coefficient, Dm2 = 0.23 ± 0.05 µm2/s, showed that corral boundaries impeded receptor diffusion 13-fold. Mathematical simulations predict the probability of G protein–coupled receptors crossing corral boundaries to be 1 in 472. Remarkably, latrunculin A, cytochalasin D, and jasplakinolide treatments altered the corral permeability. Ciliary membranes are thus partitioned into highly fluid membrane nanodomains that are delimited by filamentous actin.

scholarly journals Organized cannabinoid receptor distribution in neurons revealed by super-resolution fluorescence imaging

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Hui Li ◽  
Jie Yang ◽  
Cuiping Tian ◽  
Min Diao ◽  
Quan Wang ◽  
...  
Keyword(s):  
G Protein ◽  
Fluorescence Imaging ◽  
Cannabinoid Receptor ◽  
Super Resolution ◽  
G Protein Coupled Receptors ◽  
Cellular Functions ◽  
Cannabinoid Receptor 1 ◽  
Intracellular Organization ◽  
Dynamic Movement ◽  
G Protein Coupled

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.

EYE ON CHINA

2017 ◽  
Vol 21 (07) ◽  
pp. 4-11
Keyword(s):  
G Protein ◽  
Oral Delivery ◽  
Cell Activation ◽  
Super Resolution ◽  
Academic Paper ◽  
Conjugated Polymer Nanoparticles ◽  
Resolution Imaging ◽  
Academy Of Sciences ◽  
G Protein Coupled ◽  
Intestinal Remodeling

Chinese Researchers Develop 3D Super-Resolution Imaging at Unprecedented Depth. Scientists Advance Photosynthesis Using Conjugated Polymer Nanoparticles. Bayer Partners with Chinese Academy of Sciences to Study Photosynthetic Efficiency. Scientists Reveal How G protein-coupled Receptor Kinases Recognize and Phosphorylate G protein-coupled Receptors. China Fights Academic Paper Fraud After Peer Review Scandal. Beijing 4P Health Research Institute Initiates China’s “100K Wellness Pioneer Project” Using UniteGen and Sapientia™ Integrated Platform. SGS Expands Extractables and Leachables Testing Capabilities at its Shanghai Facility. DeltaHealth Cooperates with Cardiologists & Partners for Better Patient Care. Scientists Demonstrate RNAi as an Antiviral Immunity in Mammals. Nanodiamonds Mediate Oral Delivery of Proteins for Stem Cell Activation and Intestinal Remodeling in DrosophilaShare.

The Highly Conserved DRY Motif of Class A G Protein-Coupled Receptors: Beyond the Ground State

2006 ◽  
Vol 71 (4) ◽  
pp. 959-964 ◽  
Author(s):  
G. Enrico Rovati ◽  
Valérie Capra ◽  
Richard R. Neubig
Keyword(s):  
G Protein ◽  
Ground State ◽  
G Protein Coupled Receptors ◽  
Class A ◽  
Dry Motif ◽  
G Protein Coupled

scholarly journals Organized cannabinoid receptor distribution in neurons revealed by super-resolution fluorescence imaging

2020 ◽  
Author(s):  
Hui Li ◽  
Jie Yang ◽  
Tian Cuiping ◽  
Min Diao ◽  
Quan Wang ◽  
...  
Keyword(s):  
G Protein ◽  
Fluorescence Imaging ◽  
Cannabinoid Receptor ◽  
Super Resolution ◽  
G Protein Coupled Receptors ◽  
Cellular Functions ◽  
Cannabinoid Receptor 1 ◽  
Intracellular Organization ◽  
Dynamic Movement ◽  
G Protein Coupled

AbstractG-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 when being activated.

scholarly journals Molecular Signatures of G-Protein Coupled Receptors in Pancreatic Cancer using Super-Resolution Microscopy

2015 ◽  
Vol 108 (2) ◽  
pp. 96a
Author(s):  
Raphael Jorand ◽  
Ottavia Golfetto ◽  
Sunetra Biswas ◽  
Steven J. Tobin ◽  
Huiying Zhang ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
G Protein ◽  
Super Resolution ◽  
G Protein Coupled Receptors ◽  
Molecular Signatures ◽  
Super Resolution Microscopy ◽  
G Protein Coupled

scholarly journals Enlightening G-protein-coupled receptors on the plasma membrane using super-resolution photoactivated localization microscopy

2013 ◽  
Vol 41 (1) ◽  
pp. 191-196 ◽  
Author(s):  
Marco Scarselli ◽  
Paolo Annibale ◽  
Claudio Gerace ◽  
Aleksandra Radenovic
Keyword(s):  
Plasma Membrane ◽  
G Protein ◽  
Super Resolution ◽  
Molecular Size ◽  
G Protein Coupled Receptors ◽  
Temporal Organization ◽  
Localization Microscopy ◽  
Photoactivated Localization Microscopy ◽  
Super Resolution Microscopy ◽  
G Protein Coupled

The possibility to visualize and image the arrangement of proteins within the cell at the molecular level has always been an attraction for scientists in biological research. In particular, for signalling molecules such as GPCRs (G-protein-coupled receptors), the existence of protein aggregates such as oligomers or clusters has been the topic of extensive debate. One of the reasons for this lively argument is that the molecular size is below the diffraction-limited resolution of the conventional microscopy, precluding the direct visualization of protein super-structures. On the other hand, new super-resolution microscopy techniques, such as the PALM (photoactivated localization microscopy), allow the limit of the resolution power of conventional optics to be broken and the localization of single molecules to be determined with a precision of 10–20 nm, close to their molecular size. The application of super-resolution microscopy to study the spatial and temporal organization of GPCRs has brought new insights into receptor arrangement on the plasma membrane. Furthermore, the use of this powerful microscopy technique as a quantitative tool opens up the possibility for investigating and quantifying the number of molecules in biological assemblies and determining the protein stoichiometry in signalling complexes.

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