Quantitative Proteome Analysis in Response to Glucose Concentration in C2C12 Myotubes

Ayano Kato,Kosuke Sugiura,Iori Sakakibara,Takayuki Uchida,Takeshi Nikawa,Tomoya Fukawa,Taku Fukushima

doi:10.3390/app12031553

Abstract

Glucose is important for the maintenance of muscle function; however, it is still unclear how changes in glucose concentration affect muscle. Here, we analyzed the effect of glucose concentration on protein expression under different glucose concentration media in C2C12 myotubes. First, we performed proteome analysis in C2C12 myotubes cultured in Low (1.0 g/L), Medium (2.0 g/L), and High (4.5 g/L) glucose media. Proteome analysis revealed 113 proteins were significantly changed in group cultured in Low or Medium glucose media compared to group cultured in High glucose media. Furthermore, glycolysis, oxidative phosphorylation, and fatty acid metabolism were increased in the Medium and Low groups. Among these pathways, HK2, PFKP, NDUFA11, and FABP3 were especially upregulated proteins in Low and Medium groups. In this context, ATP production in C2C12 myotubes cultured in Low and Medium glucose media was increased. There was no significant change in myotubes morphology and myogenic differentiation factors in all groups. Finally, we examined the effect on glucose concentration in culture media on myosin isoforms expression by qRT-PCR. As a result, Myh2 and Myh4 were significantly increased in Low and Medium conditions. Altogether, Low and Medium glucose conditions induced Myh expression probably via enhancement glucose utilization.

Highlights

Muscle fibers that make up skeletal muscle are composed of slow muscle fibers, which are rich in mitochondria and predominate in aerobic energy production pathways, and fast muscle fibers, which predominate in anaerobic energy production pathways [1]
At 5 days post‐differentiation, proteome analysis was performed to examine the intracellular dynamics of the C2C12 myotubes at different glucose concentrations in the culture media (Table S1)
Consistent with the proteome observation, ATP contents were increased in myotubes under Low or Medium glucose media

Summary

Introduction

Muscle fibers that make up skeletal muscle are composed of slow muscle fibers, which are rich in mitochondria and predominate in aerobic energy production pathways, and fast muscle fibers (or intermediate types), which predominate in anaerobic energy production pathways [1]. Energy metabolism and muscle fiber type are closely related. It is well known that skeletal muscle changes its metabolic properties in response to changes in the systemic environment [2,3]. The mechanism that coordinates metabolic changes with muscle status remains unclear. Glucose is one of the essential nutrients for living organisms, and it plays a important role in energy production. Glucose is metabolized via glycolytic pathway to ATP and pyruvate, a substrate of the TCA cycle for more efficient ATP production.

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Results

Conclusion