Murine GPRC6A Mediates Cellular Responses to L-Amino Acids, but Not Osteocalcin Variants
Identifieur interne : 000596 ( Pmc/Curation ); précédent : 000595; suivant : 000597Murine GPRC6A Mediates Cellular Responses to L-Amino Acids, but Not Osteocalcin Variants
Auteurs : Patricia Rueda [Australie] ; Elizabeth Harley [France] ; Yao Lu [Australie] ; Gregory D. Stewart [Australie] ; Stewart Fabb [Australie] ; Natalie Diepenhorst [Australie] ; Béatrice Cremers [France] ; Marie-Hélène Rouillon [France] ; Isabelle Wehrle [France] ; Anne Geant [France] ; Gwladys Lamarche [France] ; Katie Leach [Australie] ; William N. Charman [Australie] ; Arthur Christopoulos [Australie] ; Roger J. Summers [Australie] ; Patrick M. Sexton [Australie] ; Christopher J. Langmead [Australie]Source :
- PLoS ONE [ 1932-6203 ] ; 2016.
Abstract
Phenotyping of
Url:
DOI: 10.1371/journal.pone.0146846
PubMed: 26785252
PubMed Central: 4718634
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<author><name sortKey="Rouillon, Marie Helene" sort="Rouillon, Marie Helene" uniqKey="Rouillon M" first="Marie-Hélène" last="Rouillon">Marie-Hélène Rouillon</name>
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<author><name sortKey="Wehrle, Isabelle" sort="Wehrle, Isabelle" uniqKey="Wehrle I" first="Isabelle" last="Wehrle">Isabelle Wehrle</name>
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<author><name sortKey="Geant, Anne" sort="Geant, Anne" uniqKey="Geant A" first="Anne" last="Geant">Anne Geant</name>
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<country xml:lang="fr">France</country>
<wicri:regionArea>Institut de Recherches Servier, Suresnes</wicri:regionArea>
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<author><name sortKey="Lamarche, Gwladys" sort="Lamarche, Gwladys" uniqKey="Lamarche G" first="Gwladys" last="Lamarche">Gwladys Lamarche</name>
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</nlm:aff>
<country xml:lang="fr">France</country>
<wicri:regionArea>Institut de Recherches Servier, Suresnes</wicri:regionArea>
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<author><name sortKey="Leach, Katie" sort="Leach, Katie" uniqKey="Leach K" first="Katie" last="Leach">Katie Leach</name>
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</nlm:aff>
<country xml:lang="fr">Australie</country>
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</affiliation>
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<author><name sortKey="Charman, William N" sort="Charman, William N" uniqKey="Charman W" first="William N." last="Charman">William N. Charman</name>
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</nlm:aff>
<country xml:lang="fr">Australie</country>
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</affiliation>
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<author><name sortKey="Christopoulos, Arthur" sort="Christopoulos, Arthur" uniqKey="Christopoulos A" first="Arthur" last="Christopoulos">Arthur Christopoulos</name>
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</nlm:aff>
<country xml:lang="fr">Australie</country>
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</affiliation>
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<author><name sortKey="Summers, Roger J" sort="Summers, Roger J" uniqKey="Summers R" first="Roger J." last="Summers">Roger J. Summers</name>
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</nlm:aff>
<country xml:lang="fr">Australie</country>
<wicri:regionArea>Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria 3052</wicri:regionArea>
</affiliation>
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<author><name sortKey="Sexton, Patrick M" sort="Sexton, Patrick M" uniqKey="Sexton P" first="Patrick M." last="Sexton">Patrick M. Sexton</name>
<affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria 3052, Australia</addr-line>
</nlm:aff>
<country xml:lang="fr">Australie</country>
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<author><name sortKey="Langmead, Christopher J" sort="Langmead, Christopher J" uniqKey="Langmead C" first="Christopher J." last="Langmead">Christopher J. Langmead</name>
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</nlm:aff>
<country xml:lang="fr">Australie</country>
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<front><div type="abstract" xml:lang="en"><p>Phenotyping of <italic>Gprc6a</italic>
KO mice has shown that this promiscuous class C G protein coupled receptor is variously involved in regulation of metabolism, inflammation and endocrine function. Such effects are described as mediated by extracellular calcium, L-amino acids, the bone-derived peptide osteocalcin (OCN) and the male hormone testosterone, introducing the concept of a bone-energy-metabolism-reproduction functional crosstalk mediated by GPRC6A. However, whilst the calcium and L-amino acid-sensing properties of GPRC6A are well established, verification of activity of osteocalcin at both human and mouse GPRC6A <italic>in vitro</italic>
has proven somewhat elusive. This study characterises the <italic>in vitro</italic>
pharmacology of mouse GPRC6A in response to its putative ligands in both recombinant and endogenous GPRC6A-expressing cells. Using cell signalling, and glucagon-like peptide (GLP)-1 and insulin release assays, our results confirm that basic L-amino acids act as agonists of the murine GPRC6A receptor in both recombinant cells and immortalised entero-endocrine and pancreatic β-cells. In contrast, our studies do not support a role for OCN as a direct ligand for mouse GPRC6A, suggesting that the reported <italic>in vivo</italic>
effects of OCN that require GPRC6A may be indirect, rather than <italic>via</italic>
direct activation of the receptor.</p>
</div>
</front>
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</author>
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<author><name sortKey="Hampson, Dr" uniqKey="Hampson D">DR Hampson</name>
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</analytic>
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<biblStruct><analytic><author><name sortKey="Christiansen, B" uniqKey="Christiansen B">B Christiansen</name>
</author>
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</TEI>
<pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">PLoS One</journal-id>
<journal-id journal-id-type="iso-abbrev">PLoS ONE</journal-id>
<journal-id journal-id-type="publisher-id">plos</journal-id>
<journal-id journal-id-type="pmc">plosone</journal-id>
<journal-title-group><journal-title>PLoS ONE</journal-title>
</journal-title-group>
<issn pub-type="epub">1932-6203</issn>
<publisher><publisher-name>Public Library of Science</publisher-name>
<publisher-loc>San Francisco, CA USA</publisher-loc>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">26785252</article-id>
<article-id pub-id-type="pmc">4718634</article-id>
<article-id pub-id-type="doi">10.1371/journal.pone.0146846</article-id>
<article-id pub-id-type="publisher-id">PONE-D-15-36847</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject>
</subj-group>
</article-categories>
<title-group><article-title>Murine GPRC6A Mediates Cellular Responses to L-Amino Acids, but Not Osteocalcin Variants</article-title>
<alt-title alt-title-type="running-head">Murine GPRC6A Responds to L-Amino Acids, but Not Osteocalcin</alt-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Rueda</surname>
<given-names>Patricia</given-names>
</name>
<xref ref-type="aff" rid="aff001"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Harley</surname>
<given-names>Elizabeth</given-names>
</name>
<xref ref-type="aff" rid="aff002"><sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Lu</surname>
<given-names>Yao</given-names>
</name>
<xref ref-type="aff" rid="aff001"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Stewart</surname>
<given-names>Gregory D.</given-names>
</name>
<xref ref-type="aff" rid="aff001"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Fabb</surname>
<given-names>Stewart</given-names>
</name>
<xref ref-type="aff" rid="aff001"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Diepenhorst</surname>
<given-names>Natalie</given-names>
</name>
<xref ref-type="aff" rid="aff001"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Cremers</surname>
<given-names>Béatrice</given-names>
</name>
<xref ref-type="aff" rid="aff002"><sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Rouillon</surname>
<given-names>Marie-Hélène</given-names>
</name>
<xref ref-type="aff" rid="aff002"><sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Wehrle</surname>
<given-names>Isabelle</given-names>
</name>
<xref ref-type="aff" rid="aff002"><sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Geant</surname>
<given-names>Anne</given-names>
</name>
<xref ref-type="aff" rid="aff002"><sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Lamarche</surname>
<given-names>Gwladys</given-names>
</name>
<xref ref-type="aff" rid="aff002"><sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Leach</surname>
<given-names>Katie</given-names>
</name>
<xref ref-type="aff" rid="aff001"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Charman</surname>
<given-names>William N.</given-names>
</name>
<xref ref-type="aff" rid="aff001"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Christopoulos</surname>
<given-names>Arthur</given-names>
</name>
<xref ref-type="aff" rid="aff001"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Summers</surname>
<given-names>Roger J.</given-names>
</name>
<xref ref-type="aff" rid="aff001"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Sexton</surname>
<given-names>Patrick M.</given-names>
</name>
<xref ref-type="aff" rid="aff001"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Langmead</surname>
<given-names>Christopher J.</given-names>
</name>
<xref ref-type="aff" rid="aff001"><sup>1</sup>
</xref>
<xref rid="cor001" ref-type="corresp">*</xref>
</contrib>
</contrib-group>
<aff id="aff001"><label>1</label>
<addr-line>Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria 3052, Australia</addr-line>
</aff>
<aff id="aff002"><label>2</label>
<addr-line>Institut de Recherches Servier, Suresnes, France</addr-line>
</aff>
<contrib-group><contrib contrib-type="editor"><name><surname>Hamel</surname>
<given-names>Frederick G</given-names>
</name>
<role>Editor</role>
<xref ref-type="aff" rid="edit1"></xref>
</contrib>
</contrib-group>
<aff id="edit1"><addr-line>Omaha Veterans Affairs Medical Center, UNITED STATES</addr-line>
</aff>
<author-notes><fn fn-type="conflict" id="coi001"><p><bold>Competing Interests: </bold>
EH, BC, M-HR, IW, AG, GL are employees of Servier. This does not alter the authors' adherence to PLOS ONE policies on sharing data and materials.</p>
</fn>
<fn fn-type="con" id="contrib001"><p>Conceived and designed the experiments: PR EH GDS CJL PMS AC RJS. Performed the experiments: PR YL GDS SF ND BC M-HR IW AG GL KL. Analyzed the data: PR YL GDS SF EH CJL. Wrote the paper: PR CJL EH WNC AC RJS PMS.</p>
</fn>
<corresp id="cor001">* E-mail: <email>chris.langmead@monash.edu</email>
</corresp>
</author-notes>
<pub-date pub-type="epub"><day>19</day>
<month>1</month>
<year>2016</year>
</pub-date>
<pub-date pub-type="collection"><year>2016</year>
</pub-date>
<volume>11</volume>
<issue>1</issue>
<elocation-id>e0146846</elocation-id>
<history><date date-type="received"><day>27</day>
<month>8</month>
<year>2015</year>
</date>
<date date-type="accepted"><day>21</day>
<month>12</month>
<year>2015</year>
</date>
</history>
<permissions><copyright-statement>© 2016 Rueda et al</copyright-statement>
<copyright-year>2016</copyright-year>
<copyright-holder>Rueda et al</copyright-holder>
<license xlink:href="http://creativecommons.org/licenses/by/4.0/"><license-p>This is an open access article distributed under the terms of the <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">Creative Commons Attribution License</ext-link>
, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.</license-p>
</license>
</permissions>
<self-uri content-type="pdf" xlink:type="simple" xlink:href="pone.0146846.pdf"></self-uri>
<abstract><p>Phenotyping of <italic>Gprc6a</italic>
KO mice has shown that this promiscuous class C G protein coupled receptor is variously involved in regulation of metabolism, inflammation and endocrine function. Such effects are described as mediated by extracellular calcium, L-amino acids, the bone-derived peptide osteocalcin (OCN) and the male hormone testosterone, introducing the concept of a bone-energy-metabolism-reproduction functional crosstalk mediated by GPRC6A. However, whilst the calcium and L-amino acid-sensing properties of GPRC6A are well established, verification of activity of osteocalcin at both human and mouse GPRC6A <italic>in vitro</italic>
has proven somewhat elusive. This study characterises the <italic>in vitro</italic>
pharmacology of mouse GPRC6A in response to its putative ligands in both recombinant and endogenous GPRC6A-expressing cells. Using cell signalling, and glucagon-like peptide (GLP)-1 and insulin release assays, our results confirm that basic L-amino acids act as agonists of the murine GPRC6A receptor in both recombinant cells and immortalised entero-endocrine and pancreatic β-cells. In contrast, our studies do not support a role for OCN as a direct ligand for mouse GPRC6A, suggesting that the reported <italic>in vivo</italic>
effects of OCN that require GPRC6A may be indirect, rather than <italic>via</italic>
direct activation of the receptor.</p>
</abstract>
<funding-group><funding-statement>Servier provided support in the form of salaries for EH, BC, M-HR, IW, AG, GL and provided research support to Monash University to fund salaries for PR, YL, GDS, SF, ND and CJL. However, Servier did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section. AC and PMS are NHMRC Principal Research Fellows.</funding-statement>
</funding-group>
<counts><fig-count count="7"></fig-count>
<table-count count="1"></table-count>
<page-count count="19"></page-count>
</counts>
<custom-meta-group><custom-meta id="data-availability"><meta-name>Data Availability</meta-name>
<meta-value>All relevant data are within the paper and its Supporting Information files.</meta-value>
</custom-meta>
</custom-meta-group>
</article-meta>
<notes><title>Data Availability</title>
<p>All relevant data are within the paper and its Supporting Information files.</p>
</notes>
</front>
</pmc>
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