A highly responsive pyruvate sensor reveals pathway-regulatory role of the mitochondrial pyruvate carrier MPC
- Center for Scientific Studies - CECs, Chile
- University of Münster, Germany
Abstract
Mitochondria generate ATP and building blocks for cell growth and regeneration, using pyruvate as the main substrate. Here we introduce PyronicSF, a user-friendly GFP-based sensor of improved dynamic range that enables real-time subcellular quantitation of mitochondrial pyruvate transport, concentration and flux. We report that cultured mouse astrocytes maintain mitochondrial pyruvate in the low micromolar range, below cytosolic pyruvate, which means that the mitochondrial pyruvate carrier MPC is poised to exert ultrasensitive control on the balance between respiration and anaplerosis/gluconeogenesis. The functionality of the sensor in living tissue is demonstrated in the brain of Drosophila melanogaster larvae. Mitochondrial subpopulations are known to coexist within a given cell, which differ in their morphology, mobility, membrane potential, and vicinity to other organelles. The present tool can be used to investigate how mitochondrial diversity relates to metabolism, to study the role of MPC in disease, and to screen for small-molecule MPC modulators.
Data availability
All data generated or analysed during this study are included in the manuscript and supporting files. Source data files have been provided for all figures.
Article and author information
Author details
Alejandro San Martín
Funding
Fondo Nacional de Desarrollo Científico y Tecnológico (11150930)
- Alejandro San Martín
Fondo Nacional de Desarrollo Científico y Tecnológico (1160317)
- L Felipe Barros
CONICYT CHILE (PB-01)
- L Felipe Barros
Deutsche Forschungsgemeinschaft (SFB1009)
- Stefanie Schirmeier
Deutsche Forschungsgemeinschaft (SCHI 1380/2)
- Stefanie Schirmeier
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. Procedures were approved by the Centro de Estudios Científicos Animal Care and Use Committee, project 1160317.
Copyright
© 2020, Arce-Molina et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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A highly responsive pyruvate sensor reveals pathway-regulatory role of the mitochondrial pyruvate carrier MPC
eLife 9:e53917.
https://doi.org/10.7554/eLife.53917
