Super-resolution microscopy compatible fluorescent probes reveal endogenous glucagon-like peptide-1 receptor distribution and dynamics
File(s)Super-resolution microscopy compatible fluorescent.pdf (10.67 MB)
Published version
Author(s)
Type
Journal Article
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.
Date Issued
2020-01-24
Date Acceptance
2019-12-27
Citation
Nature Communications, 2020, 11
ISSN
2041-1723
Publisher
Nature Research (part of Springer Nature)
Journal / Book Title
Nature Communications
Volume
11
Copyright Statement
The Author(s) 2020. This article is licensed under a Creative Commons Attribution
4.0 International License, which permits use, sharing, adaptation,
distribution and reproduction in any medium or format, as long as you give appropriate
credit to the original author(s) and the source, provide a link to the Creative Commons
license, and indicate if changes were made. The images or other third party material in this
article are included in the article’s Creative Commons license, unless indicated otherwise in
a credit line to the material. If material is not included in the article’s Creative Commons
license and your intended use is not permitted by statutory regulation or exceeds the
permitted use, you will need to obtain permission directly from the copyright holder. To
view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
4.0 International License, which permits use, sharing, adaptation,
distribution and reproduction in any medium or format, as long as you give appropriate
credit to the original author(s) and the source, provide a link to the Creative Commons
license, and indicate if changes were made. The images or other third party material in this
article are included in the article’s Creative Commons license, unless indicated otherwise in
a credit line to the material. If material is not included in the article’s Creative Commons
license and your intended use is not permitted by statutory regulation or exceeds the
permitted use, you will need to obtain permission directly from the copyright holder. To
view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
Sponsor
Medical Research Council (MRC)
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/31980626
PII: 10.1038/s41467-020-14309-w
Grant Number
MR/R010676/1
Subjects
Science & Technology
Multidisciplinary Sciences
Science & Technology - Other Topics
GLP-1 RECEPTOR
FLUOROGENIC PROBES
EXPRESSING CELLS
BETA
MOUSE
PHARMACOLOGY
BIOLOGY
RELEASE
TISSUE
ALPHA
Amino Acid Sequence
Animals
Brain
Cell Line
Fluorescent Dyes
Glucagon-Like Peptide-1 Receptor
HEK293 Cells
Human Embryonic Stem Cells
Humans
Islets of Langerhans
Mice
Mice, Knockout
Microscopy, Fluorescence, Multiphoton
Models, Molecular
Molecular Structure
Peptide Fragments
Signal Transduction
Tissue Distribution
Islets of Langerhans
Brain
Cell Line
Animals
Mice, Knockout
Humans
Mice
Peptide Fragments
Fluorescent Dyes
Microscopy, Fluorescence, Multiphoton
Signal Transduction
Molecular Structure
Amino Acid Sequence
Tissue Distribution
Models, Molecular
HEK293 Cells
Human Embryonic Stem Cells
Glucagon-Like Peptide-1 Receptor
Publication Status
Published
Coverage Spatial
England
Article Number
467
Date Publish Online
2020-01-24