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Imaging intraorganellar Ca(2+) at subcellular resolution using CEPIA
The endoplasmic reticulum (ER) and mitochondria accumulate Ca(2+) within their lumens to regulate numerous cell functions. However, determining the dynamics of intraorganellar Ca(2+) has proven to be difficult. Here we describe a family of genetically encoded Ca(2+) indicators, named calcium-measuri...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Pub. Group
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4082642/ https://www.ncbi.nlm.nih.gov/pubmed/24923787 http://dx.doi.org/10.1038/ncomms5153 |
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author | Suzuki, Junji Kanemaru, Kazunori Ishii, Kuniaki Ohkura, Masamichi Okubo, Yohei Iino, Masamitsu |
author_facet | Suzuki, Junji Kanemaru, Kazunori Ishii, Kuniaki Ohkura, Masamichi Okubo, Yohei Iino, Masamitsu |
author_sort | Suzuki, Junji |
collection | PubMed |
description | The endoplasmic reticulum (ER) and mitochondria accumulate Ca(2+) within their lumens to regulate numerous cell functions. However, determining the dynamics of intraorganellar Ca(2+) has proven to be difficult. Here we describe a family of genetically encoded Ca(2+) indicators, named calcium-measuring organelle-entrapped protein indicators (CEPIA), which can be utilized for intraorganellar Ca(2+) imaging. CEPIA, which emit green, red or blue/green fluorescence, are engineered to bind Ca(2+) at intraorganellar Ca(2+) concentrations. They can be targeted to different organelles and may be used alongside other fluorescent molecular markers, expanding the range of cell functions that can be simultaneously analysed. The spatiotemporal resolution of CEPIA makes it possible to resolve Ca(2+) import into individual mitochondria while simultaneously measuring ER and cytosolic Ca(2+). We have used these imaging capabilities to reveal differential Ca(2+) handling in individual mitochondria. CEPIA imaging is a useful new tool to further the understanding of organellar functions. |
format | Online Article Text |
id | pubmed-4082642 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Nature Pub. Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-40826422014-07-10 Imaging intraorganellar Ca(2+) at subcellular resolution using CEPIA Suzuki, Junji Kanemaru, Kazunori Ishii, Kuniaki Ohkura, Masamichi Okubo, Yohei Iino, Masamitsu Nat Commun Article The endoplasmic reticulum (ER) and mitochondria accumulate Ca(2+) within their lumens to regulate numerous cell functions. However, determining the dynamics of intraorganellar Ca(2+) has proven to be difficult. Here we describe a family of genetically encoded Ca(2+) indicators, named calcium-measuring organelle-entrapped protein indicators (CEPIA), which can be utilized for intraorganellar Ca(2+) imaging. CEPIA, which emit green, red or blue/green fluorescence, are engineered to bind Ca(2+) at intraorganellar Ca(2+) concentrations. They can be targeted to different organelles and may be used alongside other fluorescent molecular markers, expanding the range of cell functions that can be simultaneously analysed. The spatiotemporal resolution of CEPIA makes it possible to resolve Ca(2+) import into individual mitochondria while simultaneously measuring ER and cytosolic Ca(2+). We have used these imaging capabilities to reveal differential Ca(2+) handling in individual mitochondria. CEPIA imaging is a useful new tool to further the understanding of organellar functions. Nature Pub. Group 2014-06-13 /pmc/articles/PMC4082642/ /pubmed/24923787 http://dx.doi.org/10.1038/ncomms5153 Text en Copyright © 2014, Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved. http://creativecommons.org/licenses/by-nc-nd/3.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/4.0/ |
spellingShingle | Article Suzuki, Junji Kanemaru, Kazunori Ishii, Kuniaki Ohkura, Masamichi Okubo, Yohei Iino, Masamitsu Imaging intraorganellar Ca(2+) at subcellular resolution using CEPIA |
title | Imaging intraorganellar Ca(2+) at subcellular resolution using CEPIA |
title_full | Imaging intraorganellar Ca(2+) at subcellular resolution using CEPIA |
title_fullStr | Imaging intraorganellar Ca(2+) at subcellular resolution using CEPIA |
title_full_unstemmed | Imaging intraorganellar Ca(2+) at subcellular resolution using CEPIA |
title_short | Imaging intraorganellar Ca(2+) at subcellular resolution using CEPIA |
title_sort | imaging intraorganellar ca(2+) at subcellular resolution using cepia |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4082642/ https://www.ncbi.nlm.nih.gov/pubmed/24923787 http://dx.doi.org/10.1038/ncomms5153 |
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