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Label-free two-photon imaging of mitochondrial activity in murine macrophages stimulated with bacterial and viral ligands
Mitochondria are the metabolic hub of the cell, playing a central role in regulating immune responses. Dysfunction of mitochondrial reprogramming can occur during bacterial and viral infections compromising hosts’ immune signaling. Comparative evaluation of these alterations in response to bacterial...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group UK
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8263786/ https://www.ncbi.nlm.nih.gov/pubmed/34234166 http://dx.doi.org/10.1038/s41598-021-93043-9 |
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author | Allen, Christian Harry Ahmed, Duale Raiche-Tanner, Olivia Chauhan, Vinita Mostaço-Guidolin, Leila Cassol, Edana Murugkar, Sangeeta |
author_facet | Allen, Christian Harry Ahmed, Duale Raiche-Tanner, Olivia Chauhan, Vinita Mostaço-Guidolin, Leila Cassol, Edana Murugkar, Sangeeta |
author_sort | Allen, Christian Harry |
collection | PubMed |
description | Mitochondria are the metabolic hub of the cell, playing a central role in regulating immune responses. Dysfunction of mitochondrial reprogramming can occur during bacterial and viral infections compromising hosts’ immune signaling. Comparative evaluation of these alterations in response to bacterial and viral ligands can provide insights into a cell’s ability to mount pathogen-specific responses. In this study, we used two-photon excitation fluorescence (TPEF) imaging to quantify reduced nicotinamide adenine dinucleotide phosphate (NAD(P)H) and flavin adenine dinucleotide (FAD) levels in the cell and to calculate the optical redox ratio (ORR), an indicator of mitochondrial dysfunction. Analyses were performed on RAW264.7 cells and murine bone marrow derived macrophages (BMM) stimulated with bacterial (LPS) and viral (Poly(I:C)) ligands. Responses were cell type dependent, with primary cells having significantly higher levels of FAD and higher oxygen consumption rates suggesting BMM may be more dependent on mitochondrial metabolism. Our findings also suggest that FAD-TPEF intensity may be a better predictor of mitochondrial activity and localization since it demonstrates unique mitochondrial clustering patterns in LPS vs. Poly(I:C) stimulated macrophages. Collectively, we demonstrate that TPEF imaging is a powerful label-free approach for quantifying changes in mitochondrial function and organization in macrophages following bacterial and viral stimuli. |
format | Online Article Text |
id | pubmed-8263786 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-82637862021-07-09 Label-free two-photon imaging of mitochondrial activity in murine macrophages stimulated with bacterial and viral ligands Allen, Christian Harry Ahmed, Duale Raiche-Tanner, Olivia Chauhan, Vinita Mostaço-Guidolin, Leila Cassol, Edana Murugkar, Sangeeta Sci Rep Article Mitochondria are the metabolic hub of the cell, playing a central role in regulating immune responses. Dysfunction of mitochondrial reprogramming can occur during bacterial and viral infections compromising hosts’ immune signaling. Comparative evaluation of these alterations in response to bacterial and viral ligands can provide insights into a cell’s ability to mount pathogen-specific responses. In this study, we used two-photon excitation fluorescence (TPEF) imaging to quantify reduced nicotinamide adenine dinucleotide phosphate (NAD(P)H) and flavin adenine dinucleotide (FAD) levels in the cell and to calculate the optical redox ratio (ORR), an indicator of mitochondrial dysfunction. Analyses were performed on RAW264.7 cells and murine bone marrow derived macrophages (BMM) stimulated with bacterial (LPS) and viral (Poly(I:C)) ligands. Responses were cell type dependent, with primary cells having significantly higher levels of FAD and higher oxygen consumption rates suggesting BMM may be more dependent on mitochondrial metabolism. Our findings also suggest that FAD-TPEF intensity may be a better predictor of mitochondrial activity and localization since it demonstrates unique mitochondrial clustering patterns in LPS vs. Poly(I:C) stimulated macrophages. Collectively, we demonstrate that TPEF imaging is a powerful label-free approach for quantifying changes in mitochondrial function and organization in macrophages following bacterial and viral stimuli. Nature Publishing Group UK 2021-07-07 /pmc/articles/PMC8263786/ /pubmed/34234166 http://dx.doi.org/10.1038/s41598-021-93043-9 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Allen, Christian Harry Ahmed, Duale Raiche-Tanner, Olivia Chauhan, Vinita Mostaço-Guidolin, Leila Cassol, Edana Murugkar, Sangeeta Label-free two-photon imaging of mitochondrial activity in murine macrophages stimulated with bacterial and viral ligands |
title | Label-free two-photon imaging of mitochondrial activity in murine macrophages stimulated with bacterial and viral ligands |
title_full | Label-free two-photon imaging of mitochondrial activity in murine macrophages stimulated with bacterial and viral ligands |
title_fullStr | Label-free two-photon imaging of mitochondrial activity in murine macrophages stimulated with bacterial and viral ligands |
title_full_unstemmed | Label-free two-photon imaging of mitochondrial activity in murine macrophages stimulated with bacterial and viral ligands |
title_short | Label-free two-photon imaging of mitochondrial activity in murine macrophages stimulated with bacterial and viral ligands |
title_sort | label-free two-photon imaging of mitochondrial activity in murine macrophages stimulated with bacterial and viral ligands |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8263786/ https://www.ncbi.nlm.nih.gov/pubmed/34234166 http://dx.doi.org/10.1038/s41598-021-93043-9 |
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