Book chapter
Measuring Mitochondrial Pyruvate Oxidation
Techniques to Investigate Mitochondrial Function in Neurons, pp.321-338
Neuromethods, Springer New York
03/18/2017
DOI: 10.1007/978-1-4939-6890-9_16
Abstract
Pyruvate is a central metabolic intermediate and plays a prominent role in nervous system function. Neurons are highly reliant on pyruvate oxidation for maintenance of cellular energetics. Disorders in pyruvate metabolism may result in severe neurological defects. Pyruvate is imported into the mitochondrial matrix by the mitochondrial pyruvate carrier (MPC) for oxidation by the TCA cycle to support oxidative phosphorylation. Understanding the function of the MPC requires specialized methods for investigating mitochondrial pyruvate metabolism. Multiple instruments are available for measuring respiration of animal tissues, intact cells, permeabilized cells and tissue, and isolated mitochondria. Compared to other platforms, the comparative advantage of the Seahorse extracellular flux analyzer is the ability to sequentially administer treatment compounds and observe the effects on cellular respiration in a multiplexed 96-well format. Here we describe the methods and procedures for: (1) assessing mitochondrial pyruvate oxidation by intact cultured neuronal cells; (2) by mitochondria isolated from HEK293T cells, and (3) by mitochondria isolated from mouse liver, which are useful as a general model of mitochondrial function.
Details
- Title: Subtitle
- Measuring Mitochondrial Pyruvate Oxidation
- Creators
- Lawrence R Gray - Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, USAAlix A. J Rouault - Department of Physiology and Molecular Biophysics, Carver College of Medicine, University of Iowa, Iowa City, USALalita Oonthonpan - Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, USAAdam J Rauckhorst - Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, USAJulien A Sebag - Pappajohn Biomedical Institute, University of Iowa Carver College of Medicine, Iowa City, USAEric B Taylor - Pappajohn Biomedical Institute, University of Iowa Carver College of Medicine, Iowa City, USA
- Resource Type
- Book chapter
- Publication Details
- Techniques to Investigate Mitochondrial Function in Neurons, pp.321-338
- Publisher
- Springer New York; New York, NY
- Series
- Neuromethods
- DOI
- 10.1007/978-1-4939-6890-9_16
- eISSN
- 1940-6045
- ISSN
- 0893-2336
- Language
- English
- Date published
- 03/18/2017
- Academic Unit
- Iowa Neuroscience Institute; Biochemistry and Molecular Biology; Molecular Physiology and Biophysics; Fraternal Order of Eagles Diabetes Research Center
- Record Identifier
- 9984070551702771
Metrics
12 Record Views