Extracellular ATP activates store-operated Ca(2+) entry in white adipocytes: functional evidence for STIM1 and ORAI1
In the present study, we have applied ratiometric measurements of intracellular Ca(2+) concentrations ([Ca(2+)](i)) to show that extracellularly applied ATP (adenosine triphosphate) (100 µM) stimulates store-operated Ca(2+) entry (SOCE) in 3T3-L1 adipocytes. ATP produced a rapid increase in [Ca(2+)]...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Portland Press Ltd.
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5813502/ https://www.ncbi.nlm.nih.gov/pubmed/29335300 http://dx.doi.org/10.1042/BCJ20170484 |
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author | El Hachmane, Mickaël F. Ermund, Anna Brännmark, Cecilia Olofsson, Charlotta S. |
author_facet | El Hachmane, Mickaël F. Ermund, Anna Brännmark, Cecilia Olofsson, Charlotta S. |
author_sort | El Hachmane, Mickaël F. |
collection | PubMed |
description | In the present study, we have applied ratiometric measurements of intracellular Ca(2+) concentrations ([Ca(2+)](i)) to show that extracellularly applied ATP (adenosine triphosphate) (100 µM) stimulates store-operated Ca(2+) entry (SOCE) in 3T3-L1 adipocytes. ATP produced a rapid increase in [Ca(2+)](i) consisting of an initial transient elevation followed by a sustained elevated phase that could be observed only in the presence of extracellular Ca(2+). Gene expression data and [Ca(2+)](i) recordings with uridine-5′-triphosphate or with the phospholipase C (PLC) inhibitor U73122 demonstrated the involvement of purinergic P2Y2 receptors and the PLC/inositol trisphosphate pathway. The [Ca(2+)](i) elevation produced by reintroduction of a Ca(2+)-containing intracellular solution to adipocytes exposed to ATP in the absence of Ca(2+) was diminished by known SOCE antagonists. The chief molecular components of SOCE, the stromal interaction molecule 1 (STIM1) and the calcium release-activated calcium channel protein 1 (ORAI1), were detected at the mRNA and protein level. Moreover, SOCE was largely diminished in cells where STIM1 and/or ORAI1 had been silenced by small interfering (si)RNA. We conclude that extracellular ATP activates SOCE in white adipocytes, an effect predominantly mediated by STIM1 and ORAI1. |
format | Online Article Text |
id | pubmed-5813502 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Portland Press Ltd. |
record_format | MEDLINE/PubMed |
spelling | pubmed-58135022018-02-28 Extracellular ATP activates store-operated Ca(2+) entry in white adipocytes: functional evidence for STIM1 and ORAI1 El Hachmane, Mickaël F. Ermund, Anna Brännmark, Cecilia Olofsson, Charlotta S. Biochem J Research Articles In the present study, we have applied ratiometric measurements of intracellular Ca(2+) concentrations ([Ca(2+)](i)) to show that extracellularly applied ATP (adenosine triphosphate) (100 µM) stimulates store-operated Ca(2+) entry (SOCE) in 3T3-L1 adipocytes. ATP produced a rapid increase in [Ca(2+)](i) consisting of an initial transient elevation followed by a sustained elevated phase that could be observed only in the presence of extracellular Ca(2+). Gene expression data and [Ca(2+)](i) recordings with uridine-5′-triphosphate or with the phospholipase C (PLC) inhibitor U73122 demonstrated the involvement of purinergic P2Y2 receptors and the PLC/inositol trisphosphate pathway. The [Ca(2+)](i) elevation produced by reintroduction of a Ca(2+)-containing intracellular solution to adipocytes exposed to ATP in the absence of Ca(2+) was diminished by known SOCE antagonists. The chief molecular components of SOCE, the stromal interaction molecule 1 (STIM1) and the calcium release-activated calcium channel protein 1 (ORAI1), were detected at the mRNA and protein level. Moreover, SOCE was largely diminished in cells where STIM1 and/or ORAI1 had been silenced by small interfering (si)RNA. We conclude that extracellular ATP activates SOCE in white adipocytes, an effect predominantly mediated by STIM1 and ORAI1. Portland Press Ltd. 2018-02-14 2018-02-14 /pmc/articles/PMC5813502/ /pubmed/29335300 http://dx.doi.org/10.1042/BCJ20170484 Text en © 2018 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article published by Portland Press Limited on behalf of the Biochemical Society and distributed under the Creative Commons Attribution License 4.0 (CC BY) (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Research Articles El Hachmane, Mickaël F. Ermund, Anna Brännmark, Cecilia Olofsson, Charlotta S. Extracellular ATP activates store-operated Ca(2+) entry in white adipocytes: functional evidence for STIM1 and ORAI1 |
title | Extracellular ATP activates store-operated Ca(2+) entry in white adipocytes: functional evidence for STIM1 and ORAI1 |
title_full | Extracellular ATP activates store-operated Ca(2+) entry in white adipocytes: functional evidence for STIM1 and ORAI1 |
title_fullStr | Extracellular ATP activates store-operated Ca(2+) entry in white adipocytes: functional evidence for STIM1 and ORAI1 |
title_full_unstemmed | Extracellular ATP activates store-operated Ca(2+) entry in white adipocytes: functional evidence for STIM1 and ORAI1 |
title_short | Extracellular ATP activates store-operated Ca(2+) entry in white adipocytes: functional evidence for STIM1 and ORAI1 |
title_sort | extracellular atp activates store-operated ca(2+) entry in white adipocytes: functional evidence for stim1 and orai1 |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5813502/ https://www.ncbi.nlm.nih.gov/pubmed/29335300 http://dx.doi.org/10.1042/BCJ20170484 |
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