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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...

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Autores principales: Zhou, Kuo, Zhu, Xuexue, Ma, Ke, Liu, Jibin, Nürnberg, Bernd, Gawaz, Meinrad, Lang, Florian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
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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.
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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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