Monitoring and mathematical modeling of mitochondrial ATP in myotubes at single-cell level reveals two distinct populati
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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 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 * Correspondence: [email protected] 2
Received January 1, 2020; Revised April 27, 2020; Accepted May 2, 2020 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 showe
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