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Nat Commun. 2020 Jan 24;11(1):467. doi: 10.1038/s41467-020-14309-w.

Super-resolution microscopy compatible fluorescent probes reveal endogenous glucagon-like peptide-1 receptor distribution and dynamics.

Author information

1
Institute of Metabolism and Systems Research (IMSR), and Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham, Birmingham, UK.
2
Centre for Endocrinology, Diabetes and Metabolism, Birmingham Health Partners, Birmingham, UK.
3
Centre for Liver Research, College of Medical and Dental Sciences, Institute for Immunology and Immunotherapy, University of Birmingham, Birmingham, UK.
4
Genome Editing Facility, Technology Hub, University of Birmingham, Birmingham, UK.
5
Division of Diabetes, Endocrinology and Metabolism, Section of Investigative Medicine, Imperial College London, London, UK.
6
Diabetes Research Group, BC Children's Hospital Research Institute, Vancouver, BC, Canada; Department of Surgery, University of British Columbia, Vancouver, BC, Canada.
7
Centre for Cardiovascular and Metabolic Neuroscience, Department of Neuroscience, Physiology & Pharmacology, University College London, London, UK.
8
Oxford Centre for Diabetes, Endocrinology & Metabolism, University of Oxford, Oxford, UK.
9
Division of Diabetes, Endocrinology and Metabolism, Section of Cell Biology and Functional Genomics, Imperial College London, London, UK.
10
Faculty of Biology, Medicine and Health, University of Manchester, Oxford Road, Manchester, UK.
11
Wellcome Trust-MRC Institute of Metabolic Science, University of Cambridge, Cambridge, UK.
12
Department of Pediatrics, Indiana University School of Medicine, Indianapolis, IN, USA.
13
Optical Microscopy Facility, Max Planck Institute for Medical Research, Heidelberg, Germany.
14
Department of Chemical Biology, Max Planck Institute for Medical Research, Heidelberg, Germany.
15
Department of Chemical Biology, Max Planck Institute for Medical Research, Heidelberg, Germany. tom.podewin@mpimf-heidelberg.mpg.de.
16
Department of Chemical Biology, Max Planck Institute for Medical Research, Heidelberg, Germany. johannes.broichhagen@mr.mpg.de.
17
Institute of Metabolism and Systems Research (IMSR), and Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham, Birmingham, UK. d.hodson@bham.ac.uk.
18
Centre for Endocrinology, Diabetes and Metabolism, Birmingham Health Partners, Birmingham, UK. d.hodson@bham.ac.uk.

Abstract

The glucagon-like peptide-1 receptor (GLP1R) is a class B G protein-coupled receptor (GPCR) involved in metabolism. Presently, its visualization is limited to genetic manipulation, antibody detection or the use of probes that stimulate receptor activation. Herein, we present LUXendin645, a far-red fluorescent GLP1R antagonistic peptide label. LUXendin645 produces intense and specific membrane labeling throughout live and fixed tissue. GLP1R signaling can additionally be evoked when the receptor is allosterically modulated in the presence of LUXendin645. Using LUXendin645 and LUXendin651, we describe islet, brain and hESC-derived β-like cell GLP1R expression patterns, reveal higher-order GLP1R organization including membrane nanodomains, and track single receptor subpopulations. We furthermore show that the LUXendin backbone can be optimized for intravital two-photon imaging by installing a red fluorophore. Thus, our super-resolution compatible labeling probes allow visualization of endogenous GLP1R, and provide insight into class B GPCR distribution and dynamics both in vitro and in vivo.

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