Imaging mitochondrial flux in single cells with a FRET sensor for pyruvate
| Autor: | Karin Alegría, Felipe Baeza-Lehnert, Yasna Contreras-Baeza, Rocío Valdebenito, Sebastián Ceballo, Rodrigo Lerchundi, L. Felipe Barros, Alejandro San Martín |
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| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Male
Anatomy and Physiology Transcription Genetic Mouse Pyruvate transport Biosensing Techniques Mitochondrion Biochemistry Energy-Producing Processes chemistry.chemical_compound Mice Cytosol Pyruvic Acid Fluorescence Resonance Energy Transfer Glycolysis Energy-Producing Organelles Mammals Multidisciplinary Escherichia coli Proteins Brain Animal Models Cell biology Mitochondria Molecular Imaging Medicine Single-Cell Analysis Research Article Cell Physiology Science Green Fluorescent Proteins chemistry.chemical_element Calcium Biology Bioenergetics Model Organisms Bacterial Proteins Organelle Animals Humans Mice Inbred C57BL Repressor Proteins Luminescent Proteins HEK293 Cells Metabolism chemistry Pyruvic acid Physiological Processes Energy Metabolism Flux (metabolism) |
| Zdroj: | PLoS ONE, Vol 9, Iss 1, p e85780 (2014) PLoS ONE Plos One Artículos CONICYT CONICYT Chile instacron:CONICYT |
| ISSN: | 1932-6203 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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