TY - JOUR
T1 - A highly responsive pyruvate sensor reveals pathway-regulatory role of the mitochondrial pyruvate carrier MPC
AU - Arce-Molina, Robinson
AU - Cortés-Molina, Francisca
AU - Sandoval, Pamela Y.
AU - Galaz, Alex
AU - Alegría, Karin
AU - Schirmeier, Stefanie
AU - Barros, L. Felipe
AU - Martín, Alejandro San
N1 - Funding Information:
We thank Karen Everett for critical reading of the manuscript. This work was partly supported by Fondecyt grants 11150930 to ASM and 1160317 to LFB, and Deutsche Forschungsgemeinshaft (DFG) grants SFB1009 and SCHI 1380/2 to SS. The Centro de Estudios Científicos (CECs) is funded by the Chilean Government through the Centers of Excellence Basal Financing Program of CONICYT. Fondo Nacional de Desarrollo 11150930 Alejandro San Martín Científico y Tecnológico Fondo Nacional de Desarrollo 1160317 L Felipe Barros Científico y Tecnológico Comisión Nacional de Investi-gación PB-01 L Felipe Barros Científica y Tecnológica Deutsche Forschungsgemeinschaft SFB1009 Stefanie Schirmeier Deutsche Forschungsge-meinschaft SCHI 1380/2 Stefanie Schirmeier.
Funding Information:
We thank Karen Everett for critical reading of the manuscript. This work was partly supported by Fondecyt grants 11150930 to ASM and 1160317 to LFB, and Deutsche Forschungsgemeinshaft (DFG) grants SFB1009 and SCHI 1380/2 to SS. The Centro de Estudios Científicos (CECs) is funded by the Chilean Government through the Centers of Excellence Basal Financing Program of CONICYT.
Publisher Copyright:
© Arce-Molina et al.
PY - 2020/3
Y1 - 2020/3
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85082098882&partnerID=8YFLogxK
U2 - 10.7554/eLife.53917
DO - 10.7554/eLife.53917
M3 - Article
C2 - 32142409
AN - SCOPUS:85082098882
SN - 2050-084X
VL - 9
JO - eLife
JF - eLife
M1 - e53917
ER -