A New Strategy to Preserve and Assess Oxygen Consumption in Murine Tissues
Mitochondrial dysfunctions are implicated in several pathologies, such as metabolic, cardiovascular, respiratory, and neurological diseases, as well as in cancer and aging. These metabolic alterations are usually assessed in human or murine samples by mitochondrial respiratory chain enzymatic assays...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-12-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/1/109 |
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| author | Jerome Kluza Victoriane Peugnet Blanche Daunou William Laine Gwenola Kervoaze Gaëlle Rémy Anne Loyens Patrice Maboudou Quentin Fovez Corinne Grangette Isabelle Wolowczuk Philippe Gosset Guillaume Garçon Philippe Marchetti Florence Pinet Muriel Pichavant Emilie Dubois-Deruy |
| author_facet | Jerome Kluza Victoriane Peugnet Blanche Daunou William Laine Gwenola Kervoaze Gaëlle Rémy Anne Loyens Patrice Maboudou Quentin Fovez Corinne Grangette Isabelle Wolowczuk Philippe Gosset Guillaume Garçon Philippe Marchetti Florence Pinet Muriel Pichavant Emilie Dubois-Deruy |
| author_sort | Jerome Kluza |
| collection | DOAJ |
| description | Mitochondrial dysfunctions are implicated in several pathologies, such as metabolic, cardiovascular, respiratory, and neurological diseases, as well as in cancer and aging. These metabolic alterations are usually assessed in human or murine samples by mitochondrial respiratory chain enzymatic assays, by measuring the oxygen consumption of intact mitochondria isolated from tissues, or from cells obtained after physical or enzymatic disruption of the tissues. However, these methodologies do not maintain tissue multicellular organization and cell-cell interactions, known to influence mitochondrial metabolism. Here, we develop an optimal model to measure mitochondrial oxygen consumption in heart and lung tissue samples using the XF24 Extracellular Flux Analyzer (Seahorse) and discuss the advantages and limitations of this technological approach. Our results demonstrate that tissue organization, as well as mitochondrial ultrastructure and respiratory function, are preserved in heart and lung tissues freshly processed or after overnight conservation at 4 °C. Using this method, we confirmed the repeatedly reported obesity-associated mitochondrial dysfunction in the heart and extended it to the lungs. We set up and validated a new strategy to optimally assess mitochondrial function in murine tissues. As such, this method is of great potential interest for monitoring mitochondrial function in cohort samples. |
| first_indexed | 2024-03-10T03:40:10Z |
| format | Article |
| id | doaj.art-06aa0e3e0202474ea956e1aa38403525 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T03:40:10Z |
| publishDate | 2021-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-06aa0e3e0202474ea956e1aa384035252023-11-23T11:34:52ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-12-0123110910.3390/ijms23010109A New Strategy to Preserve and Assess Oxygen Consumption in Murine TissuesJerome Kluza0Victoriane Peugnet1Blanche Daunou2William Laine3Gwenola Kervoaze4Gaëlle Rémy5Anne Loyens6Patrice Maboudou7Quentin Fovez8Corinne Grangette9Isabelle Wolowczuk10Philippe Gosset11Guillaume Garçon12Philippe Marchetti13Florence Pinet14Muriel Pichavant15Emilie Dubois-Deruy16Univ. Lille, CNRS, Inserm, CHU Lille, Institut pour la Recherche sur le Cancer de Lille, UMR9020—UMR-S 1277—Canther—Cancer Heterogeneity, Plasticity and Resistance to Therapies, 59000 Lille, FranceUniv. Lille, Inserm, CHU Lille, Institut Pasteur de Lille, U1167—RID-AGE—Facteurs de Risque et Déterminants Moléculaires des Maladies Liées au Vieillissement, 59000 Lille, FranceUniv. Lille, CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, U1019—UMR9017—CIIL—Center for Infection and Immunity of Lille, 59000 Lille, FranceUniv. Lille, CNRS, Inserm, CHU Lille, Institut pour la Recherche sur le Cancer de Lille, UMR9020—UMR-S 1277—Canther—Cancer Heterogeneity, Plasticity and Resistance to Therapies, 59000 Lille, FranceUniv. Lille, CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, U1019—UMR9017—CIIL—Center for Infection and Immunity of Lille, 59000 Lille, FranceUniv. Lille, CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, U1019—UMR9017—CIIL—Center for Infection and Immunity of Lille, 59000 Lille, FranceUniv. Lille, CNRS, Inserm, CHU Lille, Institut pour la Recherche sur le Cancer de Lille, UMR9020—UMR-S 1277—Canther—Cancer Heterogeneity, Plasticity and Resistance to Therapies, 59000 Lille, FranceCentre de Bio-Pathologie, CHRU Lille, 59000 Lille, FranceUniv. Lille, CNRS, Inserm, CHU Lille, Institut pour la Recherche sur le Cancer de Lille, UMR9020—UMR-S 1277—Canther—Cancer Heterogeneity, Plasticity and Resistance to Therapies, 59000 Lille, FranceUniv. Lille, CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, U1019—UMR9017—CIIL—Center for Infection and Immunity of Lille, 59000 Lille, FranceUniv. Lille, CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, U1019—UMR9017—CIIL—Center for Infection and Immunity of Lille, 59000 Lille, FranceUniv. Lille, CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, U1019—UMR9017—CIIL—Center for Infection and Immunity of Lille, 59000 Lille, FranceUniv. Lille, CHU Lille, Institut Pasteur de Lille, ULR 4483-IMPacts de l’Environnement Chimique sur la Santé (IMPECS), 59000 Lille, FranceUniv. Lille, CNRS, Inserm, CHU Lille, Institut pour la Recherche sur le Cancer de Lille, UMR9020—UMR-S 1277—Canther—Cancer Heterogeneity, Plasticity and Resistance to Therapies, 59000 Lille, FranceUniv. Lille, Inserm, CHU Lille, Institut Pasteur de Lille, U1167—RID-AGE—Facteurs de Risque et Déterminants Moléculaires des Maladies Liées au Vieillissement, 59000 Lille, FranceUniv. Lille, CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, U1019—UMR9017—CIIL—Center for Infection and Immunity of Lille, 59000 Lille, FranceUniv. Lille, Inserm, CHU Lille, Institut Pasteur de Lille, U1167—RID-AGE—Facteurs de Risque et Déterminants Moléculaires des Maladies Liées au Vieillissement, 59000 Lille, FranceMitochondrial dysfunctions are implicated in several pathologies, such as metabolic, cardiovascular, respiratory, and neurological diseases, as well as in cancer and aging. These metabolic alterations are usually assessed in human or murine samples by mitochondrial respiratory chain enzymatic assays, by measuring the oxygen consumption of intact mitochondria isolated from tissues, or from cells obtained after physical or enzymatic disruption of the tissues. However, these methodologies do not maintain tissue multicellular organization and cell-cell interactions, known to influence mitochondrial metabolism. Here, we develop an optimal model to measure mitochondrial oxygen consumption in heart and lung tissue samples using the XF24 Extracellular Flux Analyzer (Seahorse) and discuss the advantages and limitations of this technological approach. Our results demonstrate that tissue organization, as well as mitochondrial ultrastructure and respiratory function, are preserved in heart and lung tissues freshly processed or after overnight conservation at 4 °C. Using this method, we confirmed the repeatedly reported obesity-associated mitochondrial dysfunction in the heart and extended it to the lungs. We set up and validated a new strategy to optimally assess mitochondrial function in murine tissues. As such, this method is of great potential interest for monitoring mitochondrial function in cohort samples.https://www.mdpi.com/1422-0067/23/1/109oximetryoxidative phosphorylationenergy metabolismhigh fat diet |
| spellingShingle | Jerome Kluza Victoriane Peugnet Blanche Daunou William Laine Gwenola Kervoaze Gaëlle Rémy Anne Loyens Patrice Maboudou Quentin Fovez Corinne Grangette Isabelle Wolowczuk Philippe Gosset Guillaume Garçon Philippe Marchetti Florence Pinet Muriel Pichavant Emilie Dubois-Deruy A New Strategy to Preserve and Assess Oxygen Consumption in Murine Tissues International Journal of Molecular Sciences oximetry oxidative phosphorylation energy metabolism high fat diet |
| title | A New Strategy to Preserve and Assess Oxygen Consumption in Murine Tissues |
| title_full | A New Strategy to Preserve and Assess Oxygen Consumption in Murine Tissues |
| title_fullStr | A New Strategy to Preserve and Assess Oxygen Consumption in Murine Tissues |
| title_full_unstemmed | A New Strategy to Preserve and Assess Oxygen Consumption in Murine Tissues |
| title_short | A New Strategy to Preserve and Assess Oxygen Consumption in Murine Tissues |
| title_sort | new strategy to preserve and assess oxygen consumption in murine tissues |
| topic | oximetry oxidative phosphorylation energy metabolism high fat diet |
| url | https://www.mdpi.com/1422-0067/23/1/109 |
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