Imaging mitochondrial flux in single cells with a FRET sensor for pyruvate.
Mitochondrial flux is currently accessible at low resolution. Here we introduce a genetically-encoded FRET sensor for pyruvate, and methods for quantitative measurement of pyruvate transport, pyruvate production and mitochondrial pyruvate consumption in intact individual cells at high temporal resol...
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| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2014-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0085780 |
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| _version_ | 1849330098093686784 |
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| author | Alejandro San Martín Sebastián Ceballo Felipe Baeza-Lehnert Rodrigo Lerchundi Rocío Valdebenito Yasna Contreras-Baeza Karin Alegría L Felipe Barros |
| author_facet | Alejandro San Martín Sebastián Ceballo Felipe Baeza-Lehnert Rodrigo Lerchundi Rocío Valdebenito Yasna Contreras-Baeza Karin Alegría L Felipe Barros |
| author_sort | Alejandro San Martín |
| collection | DOAJ |
| description | Mitochondrial flux is currently accessible at low resolution. Here we introduce a genetically-encoded FRET sensor for pyruvate, and methods for quantitative measurement of pyruvate transport, pyruvate production and mitochondrial pyruvate consumption in intact individual cells at high temporal resolution. In HEK293 cells, neurons and astrocytes, mitochondrial pyruvate uptake was saturated at physiological levels, showing that the metabolic rate is determined by intrinsic properties of the organelle and not by substrate availability. The potential of the sensor was further demonstrated in neurons, where mitochondrial flux was found to rise by 300% within seconds of a calcium transient triggered by a short theta burst, while glucose levels remained unaltered. In contrast, astrocytic mitochondria were insensitive to a similar calcium transient elicited by extracellular ATP. We expect the improved resolution provided by the pyruvate sensor will be of practical interest for basic and applied researchers interested in mitochondrial function. |
| format | Article |
| id | doaj-art-ebd763472cc34e4aa49c19ddf4e6e2bb |
| institution | Kabale University |
| issn | 1932-6203 |
| language | English |
| publishDate | 2014-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-ebd763472cc34e4aa49c19ddf4e6e2bb2025-08-20T03:47:05ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0191e8578010.1371/journal.pone.0085780Imaging mitochondrial flux in single cells with a FRET sensor for pyruvate.Alejandro San MartínSebastián CeballoFelipe Baeza-LehnertRodrigo LerchundiRocío ValdebenitoYasna Contreras-BaezaKarin AlegríaL Felipe BarrosMitochondrial flux is currently accessible at low resolution. Here we introduce a genetically-encoded FRET sensor for pyruvate, and methods for quantitative measurement of pyruvate transport, pyruvate production and mitochondrial pyruvate consumption in intact individual cells at high temporal resolution. In HEK293 cells, neurons and astrocytes, mitochondrial pyruvate uptake was saturated at physiological levels, showing that the metabolic rate is determined by intrinsic properties of the organelle and not by substrate availability. The potential of the sensor was further demonstrated in neurons, where mitochondrial flux was found to rise by 300% within seconds of a calcium transient triggered by a short theta burst, while glucose levels remained unaltered. In contrast, astrocytic mitochondria were insensitive to a similar calcium transient elicited by extracellular ATP. We expect the improved resolution provided by the pyruvate sensor will be of practical interest for basic and applied researchers interested in mitochondrial function.https://doi.org/10.1371/journal.pone.0085780 |
| spellingShingle | Alejandro San Martín Sebastián Ceballo Felipe Baeza-Lehnert Rodrigo Lerchundi Rocío Valdebenito Yasna Contreras-Baeza Karin Alegría L Felipe Barros Imaging mitochondrial flux in single cells with a FRET sensor for pyruvate. PLoS ONE |
| title | Imaging mitochondrial flux in single cells with a FRET sensor for pyruvate. |
| title_full | Imaging mitochondrial flux in single cells with a FRET sensor for pyruvate. |
| title_fullStr | Imaging mitochondrial flux in single cells with a FRET sensor for pyruvate. |
| title_full_unstemmed | Imaging mitochondrial flux in single cells with a FRET sensor for pyruvate. |
| title_short | Imaging mitochondrial flux in single cells with a FRET sensor for pyruvate. |
| title_sort | imaging mitochondrial flux in single cells with a fret sensor for pyruvate |
| url | https://doi.org/10.1371/journal.pone.0085780 |
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