Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: 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.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2019
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
_version_ 1783458468761436160
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
work_keys_str_mv AT waldeckweiermairmarkus developmentandapplicationofsubmitochondrialtargetedca2biosensors
AT gottschalkbenjamin developmentandapplicationofsubmitochondrialtargetedca2biosensors
AT madreitersokolowskicorinat developmentandapplicationofsubmitochondrialtargetedca2biosensors
AT ramadanimujajeta developmentandapplicationofsubmitochondrialtargetedca2biosensors
AT ziomekgabriela developmentandapplicationofsubmitochondrialtargetedca2biosensors
AT klecchristiane developmentandapplicationofsubmitochondrialtargetedca2biosensors
AT burgstallersandra developmentandapplicationofsubmitochondrialtargetedca2biosensors
AT bischofhelmut developmentandapplicationofsubmitochondrialtargetedca2biosensors
AT depaolimariar developmentandapplicationofsubmitochondrialtargetedca2biosensors
AT erogluemrah developmentandapplicationofsubmitochondrialtargetedca2biosensors
AT malliroland developmentandapplicationofsubmitochondrialtargetedca2biosensors
AT graierwolfgangf developmentandapplicationofsubmitochondrialtargetedca2biosensors