Revealing the tissue-level complexity of endogenous glucagon-like peptide-1 receptor expression and signaling
The glucagon-like peptide-1 receptor (GLP1R) is a class B G protein-coupled receptor (GPCR) involved in glucose homeostasis and food intake. GLP1R agonists (GLP1RA) are widely used in the treatment of diabetes and obesity, yet visualizing the endogenous localization, organization and dynamics of a G...
| Main Authors: | , , , , , , , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
Springer Nature
2023
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| _version_ | 1797109316410408960 |
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| author | Ast, J Nasteska, D Fine, NHF Nieves, DJ Koszegi, Z Lanoiselée, Y Cuozzo, F Viloria, K Bacon, A Luu, NT Newsome, PN Calebiro, D Owen, DM Broichhagen, J Hodson, DJ |
| author_facet | Ast, J Nasteska, D Fine, NHF Nieves, DJ Koszegi, Z Lanoiselée, Y Cuozzo, F Viloria, K Bacon, A Luu, NT Newsome, PN Calebiro, D Owen, DM Broichhagen, J Hodson, DJ |
| author_sort | Ast, J |
| collection | OXFORD |
| description | The glucagon-like peptide-1 receptor (GLP1R) is a class B G protein-coupled receptor (GPCR) involved in glucose homeostasis and food intake. GLP1R agonists (GLP1RA) are widely used in the treatment of diabetes and obesity, yet visualizing the endogenous localization, organization and dynamics of a GPCR has so far remained out of reach. In the present study, we generate mice harboring an enzyme self-label genome-edited into the endogenous Glp1r locus. We also rationally design and test various fluorescent dyes, spanning cyan to far-red wavelengths, for labeling performance in tissue. By combining these technologies, we show that endogenous GLP1R can be specifically and sensitively detected in primary tissue using multiple colors. Longitudinal analysis of GLP1R dynamics reveals heterogeneous recruitment of neighboring cell subpopulations into signaling and trafficking, with differences observed between GLP1RA classes and dual agonists. At the nanoscopic level, GLP1Rs are found to possess higher organization, undergoing GLP1RA-dependent membrane diffusion. Together, these results show the utility of enzyme self-labels for visualization and interrogation of endogenous proteins, and provide insight into the biology of a class B GPCR in primary cells and tissue. |
| first_indexed | 2024-03-07T07:40:07Z |
| format | Journal article |
| id | oxford-uuid:e34b9191-3f03-4cb2-9b13-4a0b8c77d0c1 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T07:40:07Z |
| publishDate | 2023 |
| publisher | Springer Nature |
| record_format | dspace |
| spelling | oxford-uuid:e34b9191-3f03-4cb2-9b13-4a0b8c77d0c12023-04-11T15:47:05ZRevealing the tissue-level complexity of endogenous glucagon-like peptide-1 receptor expression and signalingJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:e34b9191-3f03-4cb2-9b13-4a0b8c77d0c1EnglishSymplectic ElementsSpringer Nature2023Ast, JNasteska, DFine, NHFNieves, DJKoszegi, ZLanoiselée, YCuozzo, FViloria, KBacon, ALuu, NTNewsome, PNCalebiro, DOwen, DMBroichhagen, JHodson, DJThe glucagon-like peptide-1 receptor (GLP1R) is a class B G protein-coupled receptor (GPCR) involved in glucose homeostasis and food intake. GLP1R agonists (GLP1RA) are widely used in the treatment of diabetes and obesity, yet visualizing the endogenous localization, organization and dynamics of a GPCR has so far remained out of reach. In the present study, we generate mice harboring an enzyme self-label genome-edited into the endogenous Glp1r locus. We also rationally design and test various fluorescent dyes, spanning cyan to far-red wavelengths, for labeling performance in tissue. By combining these technologies, we show that endogenous GLP1R can be specifically and sensitively detected in primary tissue using multiple colors. Longitudinal analysis of GLP1R dynamics reveals heterogeneous recruitment of neighboring cell subpopulations into signaling and trafficking, with differences observed between GLP1RA classes and dual agonists. At the nanoscopic level, GLP1Rs are found to possess higher organization, undergoing GLP1RA-dependent membrane diffusion. Together, these results show the utility of enzyme self-labels for visualization and interrogation of endogenous proteins, and provide insight into the biology of a class B GPCR in primary cells and tissue. |
| spellingShingle | Ast, J Nasteska, D Fine, NHF Nieves, DJ Koszegi, Z Lanoiselée, Y Cuozzo, F Viloria, K Bacon, A Luu, NT Newsome, PN Calebiro, D Owen, DM Broichhagen, J Hodson, DJ Revealing the tissue-level complexity of endogenous glucagon-like peptide-1 receptor expression and signaling |
| title | Revealing the tissue-level complexity of endogenous glucagon-like peptide-1 receptor expression and signaling |
| title_full | Revealing the tissue-level complexity of endogenous glucagon-like peptide-1 receptor expression and signaling |
| title_fullStr | Revealing the tissue-level complexity of endogenous glucagon-like peptide-1 receptor expression and signaling |
| title_full_unstemmed | Revealing the tissue-level complexity of endogenous glucagon-like peptide-1 receptor expression and signaling |
| title_short | Revealing the tissue-level complexity of endogenous glucagon-like peptide-1 receptor expression and signaling |
| title_sort | revealing the tissue level complexity of endogenous glucagon like peptide 1 receptor expression and signaling |
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