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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.
Afiliación
  • Allen CH; Department of Physics, Carleton University, 1125 Colonel By Drive, Ottawa, ON, K1S 5B6, Canada.
  • Ahmed D; Department of Health Sciences, Carleton University, 1125 Colonel By Drive, Ottawa, ON, K1S 5B6, Canada.
  • Raiche-Tanner O; Department of Physics, Carleton University, 1125 Colonel By Drive, Ottawa, ON, K1S 5B6, Canada.
  • Chauhan V; Consumer and Clinical Radiation Protection Bureau, Healthy Environments and Consumer Safety Branch, Health Canada, Ottawa, K1A 0K9, Canada.
  • Mostaço-Guidolin L; Department of Systems and Computer Engineering, Carleton University, 1125 Colonel By Drive, Ottawa, ON, K1S 5B6, Canada.
  • Cassol E; Department of Health Sciences, Carleton University, 1125 Colonel By Drive, Ottawa, ON, K1S 5B6, Canada. edana.cassol@carleton.ca.
  • Murugkar S; Centre for Infection, Immunity and Inflammation, University of Ottawa, 451 Smyth Road, Ottawa, ON, K1H 8M5, Canada. edana.cassol@carleton.ca.
Sci Rep ; 11(1): 14081, 2021 07 07.
Article en En | MEDLINE | ID: mdl-34234166
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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Imagen Molecular / Macrófagos / Mitocondrias Tipo de estudio: Prognostic_studies Límite: Animals Idioma: En Revista: Sci Rep Año: 2021 Tipo del documento: Article País de afiliación: Canadá Pais de publicación: Reino Unido
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Imagen Molecular / Macrófagos / Mitocondrias Tipo de estudio: Prognostic_studies Límite: Animals Idioma: En Revista: Sci Rep Año: 2021 Tipo del documento: Article País de afiliación: Canadá Pais de publicación: Reino Unido