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Effect of MgCl(2) and GdCl(3) on ORAI1 Expression and Store-Operated Ca(2+) Entry in Megakaryocytes
In chronic kidney disease, hyperphosphatemia upregulates the Ca(2+) channel ORAI and its activating Ca(2+) sensor STIM in megakaryocytes and platelets. ORAI1 and STIM1 accomplish store-operated Ca(2+) entry (SOCE) and play a key role in platelet activation. Signaling linking phosphate to upregulatio...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8036595/ https://www.ncbi.nlm.nih.gov/pubmed/33804889 http://dx.doi.org/10.3390/ijms22073292 |
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author | Zhou, Kuo Zhu, Xuexue Ma, Ke Liu, Jibin Nürnberg, Bernd Gawaz, Meinrad Lang, Florian |
author_facet | Zhou, Kuo Zhu, Xuexue Ma, Ke Liu, Jibin Nürnberg, Bernd Gawaz, Meinrad Lang, Florian |
author_sort | Zhou, Kuo |
collection | PubMed |
description | In chronic kidney disease, hyperphosphatemia upregulates the Ca(2+) channel ORAI and its activating Ca(2+) sensor STIM in megakaryocytes and platelets. ORAI1 and STIM1 accomplish store-operated Ca(2+) entry (SOCE) and play a key role in platelet activation. Signaling linking phosphate to upregulation of ORAI1 and STIM1 includes transcription factor NFAT5 and serum and glucocorticoid-inducible kinase SGK1. In vascular smooth muscle cells, the effect of hyperphosphatemia on ORAI1/STIM1 expression and SOCE is suppressed by Mg(2+) and the calcium-sensing receptor (CaSR) agonist Gd(3+). The present study explored whether sustained exposure to Mg(2+) or Gd(3+) interferes with the phosphate-induced upregulation of NFAT5, SGK1, ORAI1,2,3, STIM1,2 and SOCE in megakaryocytes. To this end, human megakaryocytic Meg-01 cells were treated with 2 mM ß-glycerophosphate for 24 h in the absence and presence of either 1.5 mM MgCl(2) or 50 µM GdCl(3). Transcript levels were estimated utilizing q-RT-PCR, protein abundance by Western blotting, cytosolic Ca(2+) concentration ([Ca(2+)](i)) by Fura-2 fluorescence and SOCE from the increase in [Ca(2+)](i) following re-addition of extracellular Ca(2+) after store depletion with thapsigargin (1 µM). As a result, Mg(2+) and Gd(3+) upregulated CaSR and blunted or virtually abolished the phosphate-induced upregulation of NFAT5, SGK1, ORAI1,2,3, STIM1,2 and SOCE in megakaryocytes. In conclusion, Mg(2+) and the CaSR agonist Gd(3+) interfere with phosphate-induced dysregulation of [Ca(2+)](i) in megakaryocytes. |
format | Online Article Text |
id | pubmed-8036595 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-80365952021-04-12 Effect of MgCl(2) and GdCl(3) on ORAI1 Expression and Store-Operated Ca(2+) Entry in Megakaryocytes Zhou, Kuo Zhu, Xuexue Ma, Ke Liu, Jibin Nürnberg, Bernd Gawaz, Meinrad Lang, Florian Int J Mol Sci Article In chronic kidney disease, hyperphosphatemia upregulates the Ca(2+) channel ORAI and its activating Ca(2+) sensor STIM in megakaryocytes and platelets. ORAI1 and STIM1 accomplish store-operated Ca(2+) entry (SOCE) and play a key role in platelet activation. Signaling linking phosphate to upregulation of ORAI1 and STIM1 includes transcription factor NFAT5 and serum and glucocorticoid-inducible kinase SGK1. In vascular smooth muscle cells, the effect of hyperphosphatemia on ORAI1/STIM1 expression and SOCE is suppressed by Mg(2+) and the calcium-sensing receptor (CaSR) agonist Gd(3+). The present study explored whether sustained exposure to Mg(2+) or Gd(3+) interferes with the phosphate-induced upregulation of NFAT5, SGK1, ORAI1,2,3, STIM1,2 and SOCE in megakaryocytes. To this end, human megakaryocytic Meg-01 cells were treated with 2 mM ß-glycerophosphate for 24 h in the absence and presence of either 1.5 mM MgCl(2) or 50 µM GdCl(3). Transcript levels were estimated utilizing q-RT-PCR, protein abundance by Western blotting, cytosolic Ca(2+) concentration ([Ca(2+)](i)) by Fura-2 fluorescence and SOCE from the increase in [Ca(2+)](i) following re-addition of extracellular Ca(2+) after store depletion with thapsigargin (1 µM). As a result, Mg(2+) and Gd(3+) upregulated CaSR and blunted or virtually abolished the phosphate-induced upregulation of NFAT5, SGK1, ORAI1,2,3, STIM1,2 and SOCE in megakaryocytes. In conclusion, Mg(2+) and the CaSR agonist Gd(3+) interfere with phosphate-induced dysregulation of [Ca(2+)](i) in megakaryocytes. MDPI 2021-03-24 /pmc/articles/PMC8036595/ /pubmed/33804889 http://dx.doi.org/10.3390/ijms22073292 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ). |
spellingShingle | Article Zhou, Kuo Zhu, Xuexue Ma, Ke Liu, Jibin Nürnberg, Bernd Gawaz, Meinrad Lang, Florian Effect of MgCl(2) and GdCl(3) on ORAI1 Expression and Store-Operated Ca(2+) Entry in Megakaryocytes |
title | Effect of MgCl(2) and GdCl(3) on ORAI1 Expression and Store-Operated Ca(2+) Entry in Megakaryocytes |
title_full | Effect of MgCl(2) and GdCl(3) on ORAI1 Expression and Store-Operated Ca(2+) Entry in Megakaryocytes |
title_fullStr | Effect of MgCl(2) and GdCl(3) on ORAI1 Expression and Store-Operated Ca(2+) Entry in Megakaryocytes |
title_full_unstemmed | Effect of MgCl(2) and GdCl(3) on ORAI1 Expression and Store-Operated Ca(2+) Entry in Megakaryocytes |
title_short | Effect of MgCl(2) and GdCl(3) on ORAI1 Expression and Store-Operated Ca(2+) Entry in Megakaryocytes |
title_sort | effect of mgcl(2) and gdcl(3) on orai1 expression and store-operated ca(2+) entry in megakaryocytes |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8036595/ https://www.ncbi.nlm.nih.gov/pubmed/33804889 http://dx.doi.org/10.3390/ijms22073292 |
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