Label-free two-photon imaging of mitochondrial activity in murine macrophages stimulated with bacterial and viral ligands

Abstract 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...

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Autores principales:	Christian Harry Allen, Duale Ahmed, Olivia Raiche-Tanner, Vinita Chauhan, Leila Mostaço-Guidolin, Edana Cassol, Sangeeta Murugkar
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/69b38b62b31f43db85623dc9ed3fd838
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spelling	oai:doaj.org-article:69b38b62b31f43db85623dc9ed3fd8382021-12-02T18:33:57ZLabel-free two-photon imaging of mitochondrial activity in murine macrophages stimulated with bacterial and viral ligands10.1038/s41598-021-93043-92045-2322https://doaj.org/article/69b38b62b31f43db85623dc9ed3fd8382021-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-93043-9https://doaj.org/toc/2045-2322Abstract 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.Christian Harry AllenDuale AhmedOlivia Raiche-TannerVinita ChauhanLeila Mostaço-GuidolinEdana CassolSangeeta MurugkarNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Christian Harry Allen Duale Ahmed Olivia Raiche-Tanner Vinita Chauhan Leila Mostaço-Guidolin Edana Cassol Sangeeta Murugkar Label-free two-photon imaging of mitochondrial activity in murine macrophages stimulated with bacterial and viral ligands
description	Abstract 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	article
author	Christian Harry Allen Duale Ahmed Olivia Raiche-Tanner Vinita Chauhan Leila Mostaço-Guidolin Edana Cassol Sangeeta Murugkar
author_facet	Christian Harry Allen Duale Ahmed Olivia Raiche-Tanner Vinita Chauhan Leila Mostaço-Guidolin Edana Cassol Sangeeta Murugkar
author_sort	Christian Harry Allen
title	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_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_sort	label-free two-photon imaging of mitochondrial activity in murine macrophages stimulated with bacterial and viral ligands
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/69b38b62b31f43db85623dc9ed3fd838
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Label-free two-photon imaging of mitochondrial activity in murine macrophages stimulated with bacterial and viral ligands

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