Abstract
Skeletal muscle is the main tissue of lipid metabolism and accordingly is critical for homeostasis and energy production; however, the determinants of lipid accumulation in skeletal muscle are unknown. Here, we examined whether the soleus muscle (predominantly slow-twitch fibers) has a higher lipid accumulation capacity than that of the extensor digitorum longus (EDL, predominantly fast-twitch fibers) muscle in mice. Soleus and EDL muscles were harvested from male C57BL/6J mice. The mRNA levels of genes involved in fatty acid import and triglyceride synthesis and accumulation were examined in soleus and EDL muscles. The intramyocellular lipid (IMCL) droplets of muscle cross sections and isolated single fibers were visualized by staining with BODIPY493/503, and fiber types were determined by immunofluorescent detection of myosin heavy chain (MyHC) isoforms. We detected higher mRNA expression of genes related to lipid accumulation in the soleus than the EDL. We also observed a marked increase of IMCL in single fibers from the soleus, but not the EDL, after treatment with a high-fat diet plus denervation. Interestingly, greater accumulation of IMCL droplets was observed in type 2A and 2X fibers (MyHC2A- and MyHC2X-positive fibers) than type 1 fibers (MyHC1-positive fibers) in soleus muscles. These results suggest that the soleus contains more IMCL owing to the higher population of type 2A fibers, and the difference in lipid accumulation between the soleus and EDL could depend on fiber type composition.
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Acknowledgements
We are grateful to the Center for Advanced Instrumental and Educational Supports, Faculty of Agriculture, Kyushu University for the provision of expertise and technical support in the use of the confocal laser microscope Leica TCS SP8. Many thanks to Ms. Akiko Sato and Mr. Shuichi Kitaura (Kyushu University, Japan) for animal care and for help with handling chemical reagents. This work was funded by the Japan Society for the Promotion of Science (JSPS) KAKENHI #26712023 to WM, #15J07972 to YK, and #11J40017 to SS. The cost of English proofreading service was supported by Kyushu University (the program for promoting the enhancement of research universities).
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10974_2017_9468_MOESM1_ESM.pdf
ESM 1—IMCL droplets stained with BODIPY493/503 in single muscle fibers isolated from untreated mice, mice treated with either denervation or a 24-h high-fat diet alone, or mice treated with a 24-h high-fat diet plus denervation (HF + Den). Representative images from the mouse soleus (left panels) and the EDL (right panels) are shown. The images of non-treatment and HF+Den are reproduced from Fig. 4 to compare all treatments. Obvious increases of IMCL were only observed in single fibers from the soleus of HF + Den treatment (lower left panel). These obvious increases in IMCL were not found in mice treated with a HF diet or sciatic nerve denervation alone. The prevalence is shown in Table 2. The bars indicate 25 μm (PDF 11536 KB)
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Komiya, Y., Sawano, S., Mashima, D. et al. Mouse soleus (slow) muscle shows greater intramyocellular lipid droplet accumulation than EDL (fast) muscle: fiber type-specific analysis. J Muscle Res Cell Motil 38, 163–173 (2017). https://doi.org/10.1007/s10974-017-9468-6
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DOI: https://doi.org/10.1007/s10974-017-9468-6