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Differential Roles for STIM1 and STIM2 in Store-Operated Calcium Entry in Rat Neurons
The interaction between Ca(2+) sensors STIM1 and STIM2 and Ca(2+) channel-forming protein ORAI1 is a crucial element of store-operated calcium entry (SOCE) in non-excitable cells. However, the molecular mechanism of SOCE in neurons remains unclear. We addressed this issue by establishing the presenc...
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Formato: | Texto |
Lenguaje: | English |
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Public Library of Science
2011
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3082561/ https://www.ncbi.nlm.nih.gov/pubmed/21541286 http://dx.doi.org/10.1371/journal.pone.0019285 |
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author | Gruszczynska-Biegala, Joanna Pomorski, Pawel Wisniewska, Marta B. Kuznicki, Jacek |
author_facet | Gruszczynska-Biegala, Joanna Pomorski, Pawel Wisniewska, Marta B. Kuznicki, Jacek |
author_sort | Gruszczynska-Biegala, Joanna |
collection | PubMed |
description | The interaction between Ca(2+) sensors STIM1 and STIM2 and Ca(2+) channel-forming protein ORAI1 is a crucial element of store-operated calcium entry (SOCE) in non-excitable cells. However, the molecular mechanism of SOCE in neurons remains unclear. We addressed this issue by establishing the presence and function of STIM proteins. Real-time polymerase chain reaction from cortical neurons showed that these cells contain significant amounts of Stim1 and Stim2 mRNA. Thapsigargin (TG) treatment increased the amount of both endogenous STIM proteins in neuronal membrane fractions. The number of YFP-STIM1/ORAI1 and YFP-STIM2/ORAI1 complexes was also enhanced by such treatment. The differences observed in the number of STIM1 and STIM2 complexes under SOCE conditions and the differential sensitivity to SOCE inhibitors suggest their distinct roles. Endoplasmic reticulum (ER) store depletion by TG enhanced intracellular Ca(2+) levels in loaded with Fura-2 neurons transfected with YFP-STIM1 and ORAI1, but not with YFP-STIM2 and ORAI1, which correlated well with the number of complexes formed. Moreover, the SOCE inhibitors ML-9 and 2-APB reduced Ca(2+) influx in neurons expressing YFP-STIM1/ORAI1 but produced no effect in cells transfected with YFP-STIM2/ORAI1. Moreover, in neurons transfected with YFP-STIM2/ORAI1, the increase in constitutive calcium entry was greater than with YFP-STIM1/ORAI1. Our data indicate that both STIM proteins are involved in calcium homeostasis in neurons. STIM1 mainly activates SOCE, whereas STIM2 regulates resting Ca(2+) levels in the ER and Ca(2+) leakage with the additional involvement of STIM1. |
format | Text |
id | pubmed-3082561 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-30825612011-05-03 Differential Roles for STIM1 and STIM2 in Store-Operated Calcium Entry in Rat Neurons Gruszczynska-Biegala, Joanna Pomorski, Pawel Wisniewska, Marta B. Kuznicki, Jacek PLoS One Research Article The interaction between Ca(2+) sensors STIM1 and STIM2 and Ca(2+) channel-forming protein ORAI1 is a crucial element of store-operated calcium entry (SOCE) in non-excitable cells. However, the molecular mechanism of SOCE in neurons remains unclear. We addressed this issue by establishing the presence and function of STIM proteins. Real-time polymerase chain reaction from cortical neurons showed that these cells contain significant amounts of Stim1 and Stim2 mRNA. Thapsigargin (TG) treatment increased the amount of both endogenous STIM proteins in neuronal membrane fractions. The number of YFP-STIM1/ORAI1 and YFP-STIM2/ORAI1 complexes was also enhanced by such treatment. The differences observed in the number of STIM1 and STIM2 complexes under SOCE conditions and the differential sensitivity to SOCE inhibitors suggest their distinct roles. Endoplasmic reticulum (ER) store depletion by TG enhanced intracellular Ca(2+) levels in loaded with Fura-2 neurons transfected with YFP-STIM1 and ORAI1, but not with YFP-STIM2 and ORAI1, which correlated well with the number of complexes formed. Moreover, the SOCE inhibitors ML-9 and 2-APB reduced Ca(2+) influx in neurons expressing YFP-STIM1/ORAI1 but produced no effect in cells transfected with YFP-STIM2/ORAI1. Moreover, in neurons transfected with YFP-STIM2/ORAI1, the increase in constitutive calcium entry was greater than with YFP-STIM1/ORAI1. Our data indicate that both STIM proteins are involved in calcium homeostasis in neurons. STIM1 mainly activates SOCE, whereas STIM2 regulates resting Ca(2+) levels in the ER and Ca(2+) leakage with the additional involvement of STIM1. Public Library of Science 2011-04-26 /pmc/articles/PMC3082561/ /pubmed/21541286 http://dx.doi.org/10.1371/journal.pone.0019285 Text en Gruszczynska-Biegala et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Gruszczynska-Biegala, Joanna Pomorski, Pawel Wisniewska, Marta B. Kuznicki, Jacek Differential Roles for STIM1 and STIM2 in Store-Operated Calcium Entry in Rat Neurons |
title | Differential Roles for STIM1 and STIM2 in Store-Operated Calcium
Entry in Rat Neurons |
title_full | Differential Roles for STIM1 and STIM2 in Store-Operated Calcium
Entry in Rat Neurons |
title_fullStr | Differential Roles for STIM1 and STIM2 in Store-Operated Calcium
Entry in Rat Neurons |
title_full_unstemmed | Differential Roles for STIM1 and STIM2 in Store-Operated Calcium
Entry in Rat Neurons |
title_short | Differential Roles for STIM1 and STIM2 in Store-Operated Calcium
Entry in Rat Neurons |
title_sort | differential roles for stim1 and stim2 in store-operated calcium
entry in rat neurons |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3082561/ https://www.ncbi.nlm.nih.gov/pubmed/21541286 http://dx.doi.org/10.1371/journal.pone.0019285 |
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