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Exploring G Protein-Coupled Receptor Signaling in Primary Pancreatic Islets

BACKGROUND: Targeting G protein-coupled receptors (GPCRs) in pancreatic cells is feasible to modulate glucose-induced insulin secretion. Because pancreatic islets consist of several cell types and GPCRs can couple to more than one G-protein family, results obtained in pancreatic cell lines do not al...

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Autores principales: Röthe, Juliane, Kraft, Robert, Schöneberg, Torsten, Thor, Doreen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7023723/
https://www.ncbi.nlm.nih.gov/pubmed/32082084
http://dx.doi.org/10.1186/s12575-019-0116-y
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author Röthe, Juliane
Kraft, Robert
Schöneberg, Torsten
Thor, Doreen
author_facet Röthe, Juliane
Kraft, Robert
Schöneberg, Torsten
Thor, Doreen
author_sort Röthe, Juliane
collection PubMed
description BACKGROUND: Targeting G protein-coupled receptors (GPCRs) in pancreatic cells is feasible to modulate glucose-induced insulin secretion. Because pancreatic islets consist of several cell types and GPCRs can couple to more than one G-protein family, results obtained in pancreatic cell lines do not always match the response in primary cells or intact islets. Therefore, we set out to establish a protocol to analyze second messenger activation in mouse pancreatic islets. RESULTS: Activation of Gq/11-coupled receptor expressed in primary β cells increased the second messenger IP1 in an accumulation assay. Applying a Gq/11 protein inhibitor completely abolished this signal. Activation of the V1 vasopressin and ghrelin receptors, predominantly expressed in the less abundant alpha and delta cells, was not sufficient to induce a significant IP1 increase in this assay. However, fura-2-based fluorescence imaging showed calcium signals upon application of arginine vasopressin or ghrelin within intact pancreatic islets. Using the here established protocol we were also able to determine changes in intracellular cAMP levels induced by receptors coupling to Gs and Gi/o proteins. CONCLUSIONS: Detection of the second messengers IP1, cAMP, and calcium, can be used to reliably analyze GPCR activation in intact islets.
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spelling pubmed-70237232020-02-20 Exploring G Protein-Coupled Receptor Signaling in Primary Pancreatic Islets Röthe, Juliane Kraft, Robert Schöneberg, Torsten Thor, Doreen Biol Proced Online Methodology BACKGROUND: Targeting G protein-coupled receptors (GPCRs) in pancreatic cells is feasible to modulate glucose-induced insulin secretion. Because pancreatic islets consist of several cell types and GPCRs can couple to more than one G-protein family, results obtained in pancreatic cell lines do not always match the response in primary cells or intact islets. Therefore, we set out to establish a protocol to analyze second messenger activation in mouse pancreatic islets. RESULTS: Activation of Gq/11-coupled receptor expressed in primary β cells increased the second messenger IP1 in an accumulation assay. Applying a Gq/11 protein inhibitor completely abolished this signal. Activation of the V1 vasopressin and ghrelin receptors, predominantly expressed in the less abundant alpha and delta cells, was not sufficient to induce a significant IP1 increase in this assay. However, fura-2-based fluorescence imaging showed calcium signals upon application of arginine vasopressin or ghrelin within intact pancreatic islets. Using the here established protocol we were also able to determine changes in intracellular cAMP levels induced by receptors coupling to Gs and Gi/o proteins. CONCLUSIONS: Detection of the second messengers IP1, cAMP, and calcium, can be used to reliably analyze GPCR activation in intact islets. BioMed Central 2020-02-15 /pmc/articles/PMC7023723/ /pubmed/32082084 http://dx.doi.org/10.1186/s12575-019-0116-y Text en © The Author(s). 2020 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Methodology
Röthe, Juliane
Kraft, Robert
Schöneberg, Torsten
Thor, Doreen
Exploring G Protein-Coupled Receptor Signaling in Primary Pancreatic Islets
title Exploring G Protein-Coupled Receptor Signaling in Primary Pancreatic Islets
title_full Exploring G Protein-Coupled Receptor Signaling in Primary Pancreatic Islets
title_fullStr Exploring G Protein-Coupled Receptor Signaling in Primary Pancreatic Islets
title_full_unstemmed Exploring G Protein-Coupled Receptor Signaling in Primary Pancreatic Islets
title_short Exploring G Protein-Coupled Receptor Signaling in Primary Pancreatic Islets
title_sort exploring g protein-coupled receptor signaling in primary pancreatic islets
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7023723/
https://www.ncbi.nlm.nih.gov/pubmed/32082084
http://dx.doi.org/10.1186/s12575-019-0116-y
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