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Differential role of STIM1 and STIM2 during transient inward (T(in)) current generation and the maturation process in the Xenopus oocyte

BACKGROUND: The Xenopus oocyte is a useful cell model to study Ca(2+) homeostasis and cell cycle regulation, two highly interrelated processes. Here, we used antisense oligonucleotides to investigate the role in the oocyte of stromal interaction molecule (STIM) proteins that are fundamental elements...

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Autores principales: Serrano-Flores, Barbara, Garay, Edith, Vázquez-Cuevas, Francisco G, Arellano, Rogelio O
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4236480/
https://www.ncbi.nlm.nih.gov/pubmed/25399338
http://dx.doi.org/10.1186/s12899-014-0009-x
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author Serrano-Flores, Barbara
Garay, Edith
Vázquez-Cuevas, Francisco G
Arellano, Rogelio O
author_facet Serrano-Flores, Barbara
Garay, Edith
Vázquez-Cuevas, Francisco G
Arellano, Rogelio O
author_sort Serrano-Flores, Barbara
collection PubMed
description BACKGROUND: The Xenopus oocyte is a useful cell model to study Ca(2+) homeostasis and cell cycle regulation, two highly interrelated processes. Here, we used antisense oligonucleotides to investigate the role in the oocyte of stromal interaction molecule (STIM) proteins that are fundamental elements of the store-operated calcium-entry (SOCE) phenomenon, as they are both sensors for Ca(2+) concentration in the intracellular reservoirs as well as activators of the membrane channels that allow Ca(2+) influx. RESULTS: Endogenous STIM1 and STIM2 expression was demonstrated, and their synthesis was knocked down 48–72 h after injecting oocytes with specific antisense sequences. Selective elimination of their mRNA and protein expression was confirmed by PCR and Western blot analysis, and we then evaluated the effect of their absence on two endogenous responses: the opening of SOC channels elicited by G protein-coupled receptor (GPCR)-activated Ca(2+) release, and the process of maturation stimulated by progesterone. Activation of SOC channels was monitored electrically by measuring the T(in) response, a Ca(2+)-influx-dependent Cl(−) current, while maturation was assessed by germinal vesicle breakdown (GVBD) scoring and electrophysiology. CONCLUSIONS: It was found that STIM2, but not STIM1, was essential in both responses, and T(in) currents and GVBD were strongly reduced or eliminated in cells devoid of STIM2; STIM1 knockdown had no effect on the maturation process, but it reduced the T(in) response by 15 to 70%. Thus, the endogenous SOCE response in Xenopus oocytes depended mainly on STIM2, and its expression was necessary for entry into meiosis induced by progesterone. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12899-014-0009-x) contains supplementary material, which is available to authorized users.
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spelling pubmed-42364802014-11-19 Differential role of STIM1 and STIM2 during transient inward (T(in)) current generation and the maturation process in the Xenopus oocyte Serrano-Flores, Barbara Garay, Edith Vázquez-Cuevas, Francisco G Arellano, Rogelio O BMC Physiol Research Article BACKGROUND: The Xenopus oocyte is a useful cell model to study Ca(2+) homeostasis and cell cycle regulation, two highly interrelated processes. Here, we used antisense oligonucleotides to investigate the role in the oocyte of stromal interaction molecule (STIM) proteins that are fundamental elements of the store-operated calcium-entry (SOCE) phenomenon, as they are both sensors for Ca(2+) concentration in the intracellular reservoirs as well as activators of the membrane channels that allow Ca(2+) influx. RESULTS: Endogenous STIM1 and STIM2 expression was demonstrated, and their synthesis was knocked down 48–72 h after injecting oocytes with specific antisense sequences. Selective elimination of their mRNA and protein expression was confirmed by PCR and Western blot analysis, and we then evaluated the effect of their absence on two endogenous responses: the opening of SOC channels elicited by G protein-coupled receptor (GPCR)-activated Ca(2+) release, and the process of maturation stimulated by progesterone. Activation of SOC channels was monitored electrically by measuring the T(in) response, a Ca(2+)-influx-dependent Cl(−) current, while maturation was assessed by germinal vesicle breakdown (GVBD) scoring and electrophysiology. CONCLUSIONS: It was found that STIM2, but not STIM1, was essential in both responses, and T(in) currents and GVBD were strongly reduced or eliminated in cells devoid of STIM2; STIM1 knockdown had no effect on the maturation process, but it reduced the T(in) response by 15 to 70%. Thus, the endogenous SOCE response in Xenopus oocytes depended mainly on STIM2, and its expression was necessary for entry into meiosis induced by progesterone. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12899-014-0009-x) contains supplementary material, which is available to authorized users. BioMed Central 2014-11-15 /pmc/articles/PMC4236480/ /pubmed/25399338 http://dx.doi.org/10.1186/s12899-014-0009-x Text en © Serrano-Flores et al.; licensee BioMed Central Ltd. 2014 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Serrano-Flores, Barbara
Garay, Edith
Vázquez-Cuevas, Francisco G
Arellano, Rogelio O
Differential role of STIM1 and STIM2 during transient inward (T(in)) current generation and the maturation process in the Xenopus oocyte
title Differential role of STIM1 and STIM2 during transient inward (T(in)) current generation and the maturation process in the Xenopus oocyte
title_full Differential role of STIM1 and STIM2 during transient inward (T(in)) current generation and the maturation process in the Xenopus oocyte
title_fullStr Differential role of STIM1 and STIM2 during transient inward (T(in)) current generation and the maturation process in the Xenopus oocyte
title_full_unstemmed Differential role of STIM1 and STIM2 during transient inward (T(in)) current generation and the maturation process in the Xenopus oocyte
title_short Differential role of STIM1 and STIM2 during transient inward (T(in)) current generation and the maturation process in the Xenopus oocyte
title_sort differential role of stim1 and stim2 during transient inward (t(in)) current generation and the maturation process in the xenopus oocyte
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4236480/
https://www.ncbi.nlm.nih.gov/pubmed/25399338
http://dx.doi.org/10.1186/s12899-014-0009-x
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