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Quantitative Biology

ISSN 2095-4689

ISSN 2095-4697(Online)

CN 10-1028/TM

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Quant. Biol.    2020, Vol. 8 Issue (3) : 228-237    https://doi.org/10.1007/s40484-020-0211-8
RESEARCH ARTICLE
Monitoring and mathematical modeling of mitochondrial ATP in myotubes at single-cell level reveals two distinct population with different kinetics
Naoki Matsuda1, Ken-ichi Hironaka1, Masashi Fujii1,2,3, Takumi Wada1, Katsuyuki Kunida1,4, Haruki Inoue5, Miki Eto1, Daisuke Hoshino1,6, Yasuro Furuichi7, Yasuko Manabe7, Nobuharu L. Fujii7, Hiroyuki Noji8, Hiromi Imamura9, Shinya Kuroda1,2,10()
1. Department of Biological Sciences, Graduate School of Science, University of Tokyo, Tokyo 113-0033, Japan
2. Molecular Genetics Research Laboratory, Graduate School of Science, University of Tokyo, Tokyo 113-0033, Japan
3. Department of Mathematical and Life Sciences, Graduate School of Integrated Sciences for Life, Hiroshima University, Hiroshima 739-8526, Japan
4. Laboratory of Computational Biology, Graduate School of Biological Sciences, Nara Institute of Science and Technology, Nara 630-0192, Japan
5. Department of Computational Biology and Medical Sciences, Graduate School of Frontier Sciences, University of Tokyo, Tokyo 113-0033, Japan
6. Department of Engineering Science, Graduate School of Informatics and Engineering, University of Electro-Communications, Tokyo 182-8585, Japan
7. Department of Health Promotion Sciences, Graduate School of Human Health Sciences, Tokyo Metropolitan University, Tokyo 192-0397, Japan
8. Department of Applied Chemistry, Graduate School of Engineering, University of Tokyo, Tokyo 113-8656, Japan
9. Department of Functional Biology, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan
10. CREST, Japan Science and Technology Corporation, Tokyo 113-0033, Japan
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Abstract

Background: ATP is the major energy source for myotube contraction, and is quickly produced to compensate ATP consumption and to maintain sufficient ATP level. ATP is consumed mainly in cytoplasm and produced in mitochondria during myotube contraction. To understand the mechanism of ATP homeostasis during myotube contraction, it is essential to monitor mitochondrial ATP at single-cell level, and examine how ATP is produced and consumed in mitochondria.

Methods: We established C2C12 cell line stably expressing fluorescent probe of mitochondrial ATP, and induced differentiation into myotubes. We gave electric pulse stimulation to the differentiated myotubes, and measured mitochondrial ATP. We constructed mathematical model of mitochondrial ATP at single-cell level, and analyzed kinetic parameters of ATP production and consumption.

Results: We performed hierarchical clustering analysis of time course of mitochondrial ATP, which resulted in two clusters. Cluster 1 showed strong transient increase, whereas cluster 2 showed weak transient increase. Mathematical modeling at single-cell level revealed that the ATP production rate of cluster 1 was larger than that of cluster 2, and that both regulatory pathways of ATP production and consumption of cluster 1 were faster than those of cluster 2. Cluster 1 showed larger mitochondrial mass than cluster 2, suggesting that cluster 1 shows the similar property of slow muscle fibers, and cluster 2 shows the similar property of fast muscle fibers.

Conclusion: Cluster 1 showed the stronger mitochondrial ATP increase by larger ATP production rate, but not smaller consumption. Cluster 1 might reflect the larger oxidative capacity of slow muscle fiber.
Keywords mitocondrial ATP production      ATP consumption      single-cell analysis      mathematical modeling     
Corresponding Author(s): Shinya Kuroda   
Online First Date: 24 July 2020    Issue Date: 25 September 2020
 Cite this article:   
Naoki Matsuda,Ken-ichi Hironaka,Masashi Fujii, et al. Monitoring and mathematical modeling of mitochondrial ATP in myotubes at single-cell level reveals two distinct population with different kinetics[J]. Quant. Biol., 2020, 8(3): 228-237.
 URL:  
https://academic.hep.com.cn/qb/EN/10.1007/s40484-020-0211-8
https://academic.hep.com.cn/qb/EN/Y2020/V8/I3/228
Fig.1  Measurement of mitochondrial ATP level during electrical pulse stimulation in individual myotubes and analysis using mathematical model.
Fig.2  Mitochondrial ATP level in individual differentiated C2C12 myotubes during EPS.
Fig.3  Construction of a mathematical model of mitochondrial ATP in individual myotubes.
Description Cluster 1 Cluster 2
Median Q1 Q3 Median Q1 Q3
k1 (min−1) Rate constant of ATP production 7.9e–2 3.5e–2 5.2e–1 4.2e–2 1.9e–2 1.3e–1
k2 (min−1) Rate constant of ATP consumption 8.4e–3 4.0e–3 9.9e–2 1.3e–2 5.0e–3 4.7e–2
t1 (min−1) Time constant of Y 5.7e–1 2.8e–1 1.6e+0 1.2e+0 4.0e–1 2.8e+0
t2 (min−1) Time constant of Z 4.3e+0 2.6e+0 7.0e+0 5.4e+0 3.3e+0 1.5e+1
Kd1 (1) Apparent dissociation constant of Y 7.7e–1 2.1e–1 3.8e+0 1.7e+0 2.2e–1 8.9e+0
Kd2 (1) Apparent dissociation constant of Z 1.8e+0 2.1e–1 4.4e+1 2.6e+0 2.9e–1 2.5e+1
n1 (1) Hill coefficient of Y 9.5e–1 5.9e–1 1.3e+0 1.1e+0 5.0e–1 2.2e+0
n2 (1) Hill coefficient of Z 1.5e+0 8.7e–1 4.1e+0 7.9e–1 2.9e–1 1.5e+0
Ybasal (1) Initial value of Y 9.9e–2 2.6e–2 5.1e–1 2.5e–1 3.6e–2 1.5e+0
Zbasal (1) Initial value of Z 8.5e–1 2.4e–1 3.5e+0 9.3e–1 1.6e–1 4.3e+0
Tab.1  Parameters in the mathematical model
Fig.4  Characterization of model parameters.
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