1. Super-resolution microscopy compatible fluorescent probes reveal endogenous glucagon-like peptide-1 receptor distribution and dynamics
- Author
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Shugo Sasaki, Julia Ast, Daniela Nasteska, Fiona B. Ashford, Andrea Bacon, Zania Stamataki, Tom Podewin, Alejandra Tomas, Nicholas H. F. Fine, David J. Hodson, Giuseppe D'Agostino, Maria Lucey, Elisa D’Este, Christopher A. Reissaus, Amelia K. Linnemann, Stefan Trapp, Zsombor Koszegi, Johannes Broichhagen, Ben Jones, Kai Johnsson, Daniel I. Brierley, Francis C. Lynn, Benoit Hastoy, Anastasia Arvaniti, Frank Reimann, Davide Calebiro, Ast, Julia [0000-0002-0039-4762], Fine, Nicholas H F [0000-0003-2343-8534], Nasteska, Daniela [0000-0002-8996-5102], Stamataki, Zania [0000-0003-3823-4497], Sasaki, Shugo [0000-0002-3696-7809], Brierley, Daniel I [0000-0002-4360-2648], Hastoy, Benoit [0000-0003-1244-7857], D'Agostino, Giuseppe [0000-0002-3502-4251], Reimann, Frank [0000-0001-9399-6377], Lynn, Francis C [0000-0001-9318-1063], Linnemann, Amelia K [0000-0001-7356-4876], Calebiro, Davide [0000-0002-3811-1553], Trapp, Stefan [0000-0003-0665-4948], Johnsson, Kai [0000-0002-8002-1981], Podewin, Tom [0000-0002-1632-5104], Broichhagen, Johannes [0000-0003-3084-6595], Hodson, David J [0000-0002-8641-8568], Apollo - University of Cambridge Repository, Medical Research Council (MRC), Fine, Nicholas H. F. [0000-0003-2343-8534], Brierley, Daniel I. [0000-0002-4360-2648], D’Agostino, Giuseppe [0000-0002-3502-4251], Lynn, Francis C. [0000-0001-9318-1063], Linnemann, Amelia K. [0000-0001-7356-4876], Hodson, David J. [0000-0002-8641-8568], and Fine, Nicholas HF [0000-0003-2343-8534]
- Subjects
0301 basic medicine ,Models, Molecular ,genetic structures ,Lydia Becker Institute ,123 ,Human Embryonic Stem Cells ,96 ,General Physics and Astronomy ,96/33 ,MOUSE ,14 ,59 ,Optical imaging ,631/92/96 ,chemistry.chemical_compound ,Mice ,0302 clinical medicine ,Tissue Distribution ,631/80/2373/2238 ,14/19 ,Super-resolution microscopy ,EXPRESSING CELLS ,Receptor ,lcsh:Science ,Peptide sequence ,PHARMACOLOGY ,Mice, Knockout ,Multidisciplinary ,Molecular Structure ,Endocrine system and metabolic diseases ,Brain ,3. Good health ,Multidisciplinary Sciences ,631/1647/245/2226 ,Science & Technology - Other Topics ,64/60 ,Signal transduction ,psychological phenomena and processes ,hormones, hormone substitutes, and hormone antagonists ,Signal Transduction ,endocrine system ,Fluorophore ,Science ,BETA ,BIOLOGY ,behavioral disciplines and activities ,General Biochemistry, Genetics and Molecular Biology ,Article ,Glucagon-Like Peptide-1 Receptor ,38 ,Cell Line ,03 medical and health sciences ,Islets of Langerhans ,14/34 ,ResearchInstitutes_Networks_Beacons/lydia_becker_institute_of_immunology_and_inflammation ,GLP-1 RECEPTOR ,Animals ,Humans ,Amino Acid Sequence ,Glucagon-like peptide 1 receptor ,G protein-coupled receptor ,Fluorescent Dyes ,RELEASE ,Science & Technology ,45 ,FLUOROGENIC PROBES ,HEK 293 cells ,General Chemistry ,Peptide Fragments ,96/100 ,ALPHA ,030104 developmental biology ,HEK293 Cells ,Microscopy, Fluorescence, Multiphoton ,nervous system ,chemistry ,692/163/2743 ,TISSUE ,13/51 ,14/63 ,Biophysics ,lcsh:Q ,14/69 ,Chemical tools ,030217 neurology & neurosurgery - 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., Glucagon-like peptide-1 receptor is an important regulator of appetite and glucose homeostasis. Here the authors describe super-resolution microscopy and in vivo imaging compatible fluorescent probes, which reveal endogenous glucagon-like peptide-1 receptor distribution and dynamics in islets and brain.
- Published
- 2020
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