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Development and Application of Sub-Mitochondrial Targeted Ca(2 +) Biosensors
Mitochondrial Ca(2+) uptake into the mitochondrial matrix is a well-established mechanism. However, the sub-organellar Ca(2+) kinetics remain elusive. In the present work we identified novel site-specific targeting sequences for the intermembrane space (IMS) and the cristae lumen (CL). We used these...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6788349/ https://www.ncbi.nlm.nih.gov/pubmed/31636543 http://dx.doi.org/10.3389/fncel.2019.00449 |
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author | Waldeck-Weiermair, Markus Gottschalk, Benjamin Madreiter-Sokolowski, Corina T. Ramadani-Muja, Jeta Ziomek, Gabriela Klec, Christiane Burgstaller, Sandra Bischof, Helmut Depaoli, Maria R. Eroglu, Emrah Malli, Roland Graier, Wolfgang F. |
author_facet | Waldeck-Weiermair, Markus Gottschalk, Benjamin Madreiter-Sokolowski, Corina T. Ramadani-Muja, Jeta Ziomek, Gabriela Klec, Christiane Burgstaller, Sandra Bischof, Helmut Depaoli, Maria R. Eroglu, Emrah Malli, Roland Graier, Wolfgang F. |
author_sort | Waldeck-Weiermair, Markus |
collection | PubMed |
description | Mitochondrial Ca(2+) uptake into the mitochondrial matrix is a well-established mechanism. However, the sub-organellar Ca(2+) kinetics remain elusive. In the present work we identified novel site-specific targeting sequences for the intermembrane space (IMS) and the cristae lumen (CL). We used these novel targeting peptides to develop green- and red- Ca(2+) biosensors targeted to the IMS and to the CL. Based on their distinctive spectral properties, and comparable sensitivities these novel constructs were suitable to visualize Ca(2+)-levels in various (sub) compartments in a multi-chromatic manner. Functional studies that applied these new biosensors revealed that knockdown of MCU and EMRE yielded elevated Ca(2+) levels inside the CL but not the IMS in response to IP(3)-generating agonists. Knockdown of VDAC1, however, strongly impeded the transfer of Ca(2+) through the OMM while the cytosolic Ca(2+) signal remained unchanged. The novel sub-mitochondrially targeted Ca(2+) biosensors proved to be suitable for Ca(2+) imaging with high spatial and temporal resolution in a multi-chromatic manner allowing simultaneous measurements. These informative biosensors will facilitate efforts to dissect the complex sub-mitochondrial Ca(2+) signaling under (patho)physiological conditions. |
format | Online Article Text |
id | pubmed-6788349 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-67883492019-10-21 Development and Application of Sub-Mitochondrial Targeted Ca(2 +) Biosensors Waldeck-Weiermair, Markus Gottschalk, Benjamin Madreiter-Sokolowski, Corina T. Ramadani-Muja, Jeta Ziomek, Gabriela Klec, Christiane Burgstaller, Sandra Bischof, Helmut Depaoli, Maria R. Eroglu, Emrah Malli, Roland Graier, Wolfgang F. Front Cell Neurosci Neuroscience Mitochondrial Ca(2+) uptake into the mitochondrial matrix is a well-established mechanism. However, the sub-organellar Ca(2+) kinetics remain elusive. In the present work we identified novel site-specific targeting sequences for the intermembrane space (IMS) and the cristae lumen (CL). We used these novel targeting peptides to develop green- and red- Ca(2+) biosensors targeted to the IMS and to the CL. Based on their distinctive spectral properties, and comparable sensitivities these novel constructs were suitable to visualize Ca(2+)-levels in various (sub) compartments in a multi-chromatic manner. Functional studies that applied these new biosensors revealed that knockdown of MCU and EMRE yielded elevated Ca(2+) levels inside the CL but not the IMS in response to IP(3)-generating agonists. Knockdown of VDAC1, however, strongly impeded the transfer of Ca(2+) through the OMM while the cytosolic Ca(2+) signal remained unchanged. The novel sub-mitochondrially targeted Ca(2+) biosensors proved to be suitable for Ca(2+) imaging with high spatial and temporal resolution in a multi-chromatic manner allowing simultaneous measurements. These informative biosensors will facilitate efforts to dissect the complex sub-mitochondrial Ca(2+) signaling under (patho)physiological conditions. Frontiers Media S.A. 2019-10-04 /pmc/articles/PMC6788349/ /pubmed/31636543 http://dx.doi.org/10.3389/fncel.2019.00449 Text en Copyright © 2019 Waldeck-Weiermair, Gottschalk, Madreiter-Sokolowski, Ramadani-Muja, Ziomek, Klec, Burgstaller, Bischof, Depaoli, Eroglu, Malli and Graier. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Neuroscience Waldeck-Weiermair, Markus Gottschalk, Benjamin Madreiter-Sokolowski, Corina T. Ramadani-Muja, Jeta Ziomek, Gabriela Klec, Christiane Burgstaller, Sandra Bischof, Helmut Depaoli, Maria R. Eroglu, Emrah Malli, Roland Graier, Wolfgang F. Development and Application of Sub-Mitochondrial Targeted Ca(2 +) Biosensors |
title | Development and Application of Sub-Mitochondrial Targeted Ca(2 +) Biosensors |
title_full | Development and Application of Sub-Mitochondrial Targeted Ca(2 +) Biosensors |
title_fullStr | Development and Application of Sub-Mitochondrial Targeted Ca(2 +) Biosensors |
title_full_unstemmed | Development and Application of Sub-Mitochondrial Targeted Ca(2 +) Biosensors |
title_short | Development and Application of Sub-Mitochondrial Targeted Ca(2 +) Biosensors |
title_sort | development and application of sub-mitochondrial targeted ca(2 +) biosensors |
topic | Neuroscience |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6788349/ https://www.ncbi.nlm.nih.gov/pubmed/31636543 http://dx.doi.org/10.3389/fncel.2019.00449 |
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