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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Autores principales:	Alejandro San Martín, Sebastián Ceballo, Felipe Baeza-Lehnert, Rodrigo Lerchundi, Rocío Valdebenito, Yasna Contreras-Baeza, Karin Alegría, L Felipe Barros
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/ebd763472cc34e4aa49c19ddf4e6e2bb
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id	oai:doaj.org-article:ebd763472cc34e4aa49c19ddf4e6e2bb
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spelling	oai:doaj.org-article:ebd763472cc34e4aa49c19ddf4e6e2bb2021-11-18T08:36:58ZImaging mitochondrial flux in single cells with a FRET sensor for pyruvate.1932-620310.1371/journal.pone.0085780https://doaj.org/article/ebd763472cc34e4aa49c19ddf4e6e2bb2014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24465702/?tool=EBIhttps://doaj.org/toc/1932-6203Mitochondrial 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.Alejandro San MartínSebastián CeballoFelipe Baeza-LehnertRodrigo LerchundiRocío ValdebenitoYasna Contreras-BaezaKarin AlegríaL Felipe BarrosPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 1, p e85780 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q 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.
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
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
title	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_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_sort	imaging mitochondrial flux in single cells with a fret sensor for pyruvate.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/ebd763472cc34e4aa49c19ddf4e6e2bb
work_keys_str_mv	AT alejandrosanmartin imagingmitochondrialfluxinsinglecellswithafretsensorforpyruvate AT sebastianceballo imagingmitochondrialfluxinsinglecellswithafretsensorforpyruvate AT felipebaezalehnert imagingmitochondrialfluxinsinglecellswithafretsensorforpyruvate AT rodrigolerchundi imagingmitochondrialfluxinsinglecellswithafretsensorforpyruvate AT rociovaldebenito imagingmitochondrialfluxinsinglecellswithafretsensorforpyruvate AT yasnacontrerasbaeza imagingmitochondrialfluxinsinglecellswithafretsensorforpyruvate AT karinalegria imagingmitochondrialfluxinsinglecellswithafretsensorforpyruvate AT lfelipebarros imagingmitochondrialfluxinsinglecellswithafretsensorforpyruvate
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Imaging mitochondrial flux in single cells with a FRET sensor for pyruvate.

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