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Mechanical characterization of isolated mitochondria under conditions of oxidative stress

Mechanical properties have been proven to be a pivotal parameter to enhance our understanding of living systems. While research during the last decades focused on cells and tissues, little is known about the role of organelle mechanics in cell function. Here, mitochondria are of specific interest du...

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Autores principales: Komaragiri, Yesaswini, Panhwar, Muzaffar H., Fregin, Bob, Jagirdar, Gayatri, Wolke, Carmen, Spiegler, Stefanie, Otto, Oliver
Formato: Online Artículo Texto
Lenguaje:English
Publicado: AIP Publishing LLC 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9674388/
https://www.ncbi.nlm.nih.gov/pubmed/36406339
http://dx.doi.org/10.1063/5.0111581
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author Komaragiri, Yesaswini
Panhwar, Muzaffar H.
Fregin, Bob
Jagirdar, Gayatri
Wolke, Carmen
Spiegler, Stefanie
Otto, Oliver
author_facet Komaragiri, Yesaswini
Panhwar, Muzaffar H.
Fregin, Bob
Jagirdar, Gayatri
Wolke, Carmen
Spiegler, Stefanie
Otto, Oliver
author_sort Komaragiri, Yesaswini
collection PubMed
description Mechanical properties have been proven to be a pivotal parameter to enhance our understanding of living systems. While research during the last decades focused on cells and tissues, little is known about the role of organelle mechanics in cell function. Here, mitochondria are of specific interest due to their involvement in numerous physiological and pathological processes, e.g., in the production and homeostasis of reactive oxygen species (ROS). Using real-time fluorescence and deformability cytometry, we present a microfluidic technology that is capable to determine the mechanical properties of individual mitochondria at a throughput exceeding 100 organelles per second. Our data on several thousands of viable mitochondria isolated from rat C6 glial cells yield a homogenous population with a median deformation that scales with the applied hydrodynamic stress. In two proof-of-principle studies, we investigated the impact of exogenously and endogenously produced ROS on mitochondria mechanics. Exposing C6 cells to hydrogen peroxide (H(2)O(2)) triggers superoxide production and leads to a reduction in mitochondria size while deformation is increased. In a second study, we focused on the knockout of tafazzin, which has been associated with impaired remodeling of the mitochondrial membrane and elevated levels of ROS. Interestingly, our results reveal the same mechanical alterations as observed after the exposure to H(2)O(2), which points to a unified biophysical mechanism of how mitochondria respond to the presence of oxidative stress. In summary, we introduce high-throughput mechanical phenotyping into the field of organelle biology with potential applications for understanding sub-cellular dynamics that have not been accessible before.
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spelling pubmed-96743882022-11-19 Mechanical characterization of isolated mitochondria under conditions of oxidative stress Komaragiri, Yesaswini Panhwar, Muzaffar H. Fregin, Bob Jagirdar, Gayatri Wolke, Carmen Spiegler, Stefanie Otto, Oliver Biomicrofluidics Regular Articles Mechanical properties have been proven to be a pivotal parameter to enhance our understanding of living systems. While research during the last decades focused on cells and tissues, little is known about the role of organelle mechanics in cell function. Here, mitochondria are of specific interest due to their involvement in numerous physiological and pathological processes, e.g., in the production and homeostasis of reactive oxygen species (ROS). Using real-time fluorescence and deformability cytometry, we present a microfluidic technology that is capable to determine the mechanical properties of individual mitochondria at a throughput exceeding 100 organelles per second. Our data on several thousands of viable mitochondria isolated from rat C6 glial cells yield a homogenous population with a median deformation that scales with the applied hydrodynamic stress. In two proof-of-principle studies, we investigated the impact of exogenously and endogenously produced ROS on mitochondria mechanics. Exposing C6 cells to hydrogen peroxide (H(2)O(2)) triggers superoxide production and leads to a reduction in mitochondria size while deformation is increased. In a second study, we focused on the knockout of tafazzin, which has been associated with impaired remodeling of the mitochondrial membrane and elevated levels of ROS. Interestingly, our results reveal the same mechanical alterations as observed after the exposure to H(2)O(2), which points to a unified biophysical mechanism of how mitochondria respond to the presence of oxidative stress. In summary, we introduce high-throughput mechanical phenotyping into the field of organelle biology with potential applications for understanding sub-cellular dynamics that have not been accessible before. AIP Publishing LLC 2022-11-17 /pmc/articles/PMC9674388/ /pubmed/36406339 http://dx.doi.org/10.1063/5.0111581 Text en © 2022 Author(s). https://creativecommons.org/licenses/by/4.0/All article content, except where otherwise noted, is licensed under a Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ).
spellingShingle Regular Articles
Komaragiri, Yesaswini
Panhwar, Muzaffar H.
Fregin, Bob
Jagirdar, Gayatri
Wolke, Carmen
Spiegler, Stefanie
Otto, Oliver
Mechanical characterization of isolated mitochondria under conditions of oxidative stress
title Mechanical characterization of isolated mitochondria under conditions of oxidative stress
title_full Mechanical characterization of isolated mitochondria under conditions of oxidative stress
title_fullStr Mechanical characterization of isolated mitochondria under conditions of oxidative stress
title_full_unstemmed Mechanical characterization of isolated mitochondria under conditions of oxidative stress
title_short Mechanical characterization of isolated mitochondria under conditions of oxidative stress
title_sort mechanical characterization of isolated mitochondria under conditions of oxidative stress
topic Regular Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9674388/
https://www.ncbi.nlm.nih.gov/pubmed/36406339
http://dx.doi.org/10.1063/5.0111581
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