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In vivo fluorescence lifetime imaging of macrophage intracellular metabolism during wound responses in zebrafish

The function of macrophages in vitro is linked to their metabolic rewiring. However, macrophage metabolism remains poorly characterized in situ. Here, we used two-photon intensity and lifetime imaging of autofluorescent metabolic coenzymes, nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) and...

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Autores principales: Miskolci, Veronika, Tweed, Kelsey E, Lasarev, Michael R, Britt, Emily C, Walsh, Alex J, Zimmerman, Landon J, McDougal, Courtney E, Cronan, Mark R, Fan, Jing, Sauer, John-Demian, Skala, Melissa C, Huttenlocher, Anna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8871371/
https://www.ncbi.nlm.nih.gov/pubmed/35200139
http://dx.doi.org/10.7554/eLife.66080
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author Miskolci, Veronika
Tweed, Kelsey E
Lasarev, Michael R
Britt, Emily C
Walsh, Alex J
Zimmerman, Landon J
McDougal, Courtney E
Cronan, Mark R
Fan, Jing
Sauer, John-Demian
Skala, Melissa C
Huttenlocher, Anna
author_facet Miskolci, Veronika
Tweed, Kelsey E
Lasarev, Michael R
Britt, Emily C
Walsh, Alex J
Zimmerman, Landon J
McDougal, Courtney E
Cronan, Mark R
Fan, Jing
Sauer, John-Demian
Skala, Melissa C
Huttenlocher, Anna
author_sort Miskolci, Veronika
collection PubMed
description The function of macrophages in vitro is linked to their metabolic rewiring. However, macrophage metabolism remains poorly characterized in situ. Here, we used two-photon intensity and lifetime imaging of autofluorescent metabolic coenzymes, nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) and flavin adenine dinucleotide (FAD), to assess the metabolism of macrophages in the wound microenvironment. Inhibiting glycolysis reduced NAD(P)H mean lifetime and made the intracellular redox state of macrophages more oxidized, as indicated by reduced optical redox ratio. We found that TNFα+ macrophages had lower NAD(P)H mean lifetime and were more oxidized compared to TNFα− macrophages. Both infection and thermal injury induced a macrophage population with a more oxidized redox state in wounded tissues. Kinetic analysis detected temporal changes in the optical redox ratio during tissue repair, revealing a shift toward a more reduced redox state over time. Metformin reduced TNFα+ wound macrophages, made intracellular redox state more reduced and improved tissue repair. By contrast, depletion of STAT6 increased TNFα+ wound macrophages, made redox state more oxidized and impaired regeneration. Our findings suggest that autofluorescence of NAD(P)H and FAD is sensitive to dynamic changes in intracellular metabolism in tissues and can be used to probe the temporal and spatial regulation of macrophage metabolism during tissue damage and repair.
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spelling pubmed-88713712022-02-25 In vivo fluorescence lifetime imaging of macrophage intracellular metabolism during wound responses in zebrafish Miskolci, Veronika Tweed, Kelsey E Lasarev, Michael R Britt, Emily C Walsh, Alex J Zimmerman, Landon J McDougal, Courtney E Cronan, Mark R Fan, Jing Sauer, John-Demian Skala, Melissa C Huttenlocher, Anna eLife Cell Biology The function of macrophages in vitro is linked to their metabolic rewiring. However, macrophage metabolism remains poorly characterized in situ. Here, we used two-photon intensity and lifetime imaging of autofluorescent metabolic coenzymes, nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) and flavin adenine dinucleotide (FAD), to assess the metabolism of macrophages in the wound microenvironment. Inhibiting glycolysis reduced NAD(P)H mean lifetime and made the intracellular redox state of macrophages more oxidized, as indicated by reduced optical redox ratio. We found that TNFα+ macrophages had lower NAD(P)H mean lifetime and were more oxidized compared to TNFα− macrophages. Both infection and thermal injury induced a macrophage population with a more oxidized redox state in wounded tissues. Kinetic analysis detected temporal changes in the optical redox ratio during tissue repair, revealing a shift toward a more reduced redox state over time. Metformin reduced TNFα+ wound macrophages, made intracellular redox state more reduced and improved tissue repair. By contrast, depletion of STAT6 increased TNFα+ wound macrophages, made redox state more oxidized and impaired regeneration. Our findings suggest that autofluorescence of NAD(P)H and FAD is sensitive to dynamic changes in intracellular metabolism in tissues and can be used to probe the temporal and spatial regulation of macrophage metabolism during tissue damage and repair. eLife Sciences Publications, Ltd 2022-02-24 /pmc/articles/PMC8871371/ /pubmed/35200139 http://dx.doi.org/10.7554/eLife.66080 Text en © 2022, Miskolci et al https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Cell Biology
Miskolci, Veronika
Tweed, Kelsey E
Lasarev, Michael R
Britt, Emily C
Walsh, Alex J
Zimmerman, Landon J
McDougal, Courtney E
Cronan, Mark R
Fan, Jing
Sauer, John-Demian
Skala, Melissa C
Huttenlocher, Anna
In vivo fluorescence lifetime imaging of macrophage intracellular metabolism during wound responses in zebrafish
title In vivo fluorescence lifetime imaging of macrophage intracellular metabolism during wound responses in zebrafish
title_full In vivo fluorescence lifetime imaging of macrophage intracellular metabolism during wound responses in zebrafish
title_fullStr In vivo fluorescence lifetime imaging of macrophage intracellular metabolism during wound responses in zebrafish
title_full_unstemmed In vivo fluorescence lifetime imaging of macrophage intracellular metabolism during wound responses in zebrafish
title_short In vivo fluorescence lifetime imaging of macrophage intracellular metabolism during wound responses in zebrafish
title_sort in vivo fluorescence lifetime imaging of macrophage intracellular metabolism during wound responses in zebrafish
topic Cell Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8871371/
https://www.ncbi.nlm.nih.gov/pubmed/35200139
http://dx.doi.org/10.7554/eLife.66080
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