Journal of Muscle Research and Cell Motility

, Volume 38, Issue 2, pp 163–173 | Cite as

Mouse soleus (slow) muscle shows greater intramyocellular lipid droplet accumulation than EDL (fast) muscle: fiber type-specific analysis

  • Yusuke Komiya
  • Shoko Sawano
  • Daisuke Mashima
  • Riho Ichitsubo
  • Mako Nakamura
  • Ryuichi Tatsumi
  • Yoshihide Ikeuchi
  • Wataru Mizunoya
Original Article

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.

Keywords

Skeletal muscle Intramyocellular lipid (IMCL) Muscle fiber type Slow-twitch fiber Fast-twitch fiber 

Notes

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).

Supplementary material

10974_2017_9468_MOESM1_ESM.pdf (11.3 mb)
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)

References

  1. Behnke BJ, McDonough P, Padilla DJ, Musch TI, Poole DC (2003) Oxygen exchange profile in rat muscles of contrasting fibre types. J Physiol 549:597–605. doi: 10.1113/jphysiol.2002.035915 CrossRefPubMedPubMedCentralGoogle Scholar
  2. Bergouignan A, Trudel G, Simon C, Chopard A, Schoeller DA, Momken I, Votruba SB, Desage M, Burdge GC, Gauquelin-Koch G, Normand S, Blanc S (2009) Physical inactivity differentially alters dietary oleate and palmitate trafficking. Diabetes 58:367–376. doi: 10.2337/db08-0263 CrossRefPubMedPubMedCentralGoogle Scholar
  3. Bloemberg D, Quadrilatero J (2012) Rapid determination of myosin heavy chain expression in rat, mouse, and human skeletal muscle using multicolor immunofluorescence analysis. PLoS ONE 7:e35273. doi: 10.1371/journal.pone.0035273 CrossRefPubMedPubMedCentralGoogle Scholar
  4. Bosma M (2016) Lipid droplet dynamics in skeletal muscle. Exp Cell Res 340:180–186. doi: 10.1016/j.yexcr.2015.10.023 CrossRefPubMedGoogle Scholar
  5. Cree MG, Paddon-Jones D, Newcomer BR, Ronsen O, Aarsland A, Wolfe RR, Ferrando A (2010) Twenty-eight-day bed rest with hypercortisolemia induces peripheral insulin resistance and increases intramuscular triglycerides. Metabolism 59:703–710. doi: 10.1016/j.metabol.2009.09.014 CrossRefPubMedGoogle Scholar
  6. Dickinson JM, Lee JD, Sullivan BE, Harber MP, Trappe SW, Trappe TA (2010) A new method to study in vivo protein synthesis in slow- and fast-twitch muscle fibers and initial measurements in humans. J Appl Physiol (1985) 108:1410–1416 doi: 10.1152/japplphysiol.00905.2009 CrossRefPubMedCentralGoogle Scholar
  7. Essen-Gustavsson B, Karlsson A, Lundstrom K, Enfalt AC (1994) Intramuscular fat and muscle fibre lipid contents in halothane-gene-free pigs fed high or low protein diets and its relation to meat quality. Meat Sci 38:269–277. doi: 10.1016/0309-1740(94)90116-3 CrossRefPubMedGoogle Scholar
  8. Folch J, Lees M, Sloane Stanley GH (1957) A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem 226:497–509PubMedGoogle Scholar
  9. Gauthier GF (1979) Ultrastructural identification of muscle fiber types by immunocytochemistry. J Cell Biol 82:391–400CrossRefPubMedGoogle Scholar
  10. Gueugneau M, Coudy-Gandilhon C, Theron L, Meunier B, Barboiron C, Combaret L, Taillandier D, Polge C, Attaix D, Picard B, Verney J, Roche F, Feasson L, Barthelemy JC, Bechet D (2015) Skeletal muscle lipid content and oxidative activity in relation to muscle fiber type in aging and metabolic syndrome. J Gerontol A Biol Sci Med Sci 70:566–576. doi: 10.1093/gerona/glu086 CrossRefPubMedGoogle Scholar
  11. Guo Z, Burguera B, Jensen MD (2000) Kinetics of intramuscular triglyceride fatty acids in exercising humans. J Appl Physiol (1985) 89:2057–2064Google Scholar
  12. Komiya Y, Anderson JE, Akahoshi M, Nakamura M, Tatsumi R, Ikeuchi Y, Mizunoya W (2015) Protocol for rat single muscle fiber isolation and culture. Anal Biochem 482:22–24. doi: 10.1016/j.ab.2015.03.034 CrossRefPubMedGoogle Scholar
  13. Louzada RA, Santos MC, Cavalcanti-de-Albuquerque JP, Rangel IF, Ferreira AC, Galina A, Werneck-de-Castro JP, Carvalho DP (2014) Type 2 iodothyronine deiodinase is upregulated in rat slow- and fast-twitch skeletal muscle during cold exposure. Am J Physiol Endocrinol Metab 307:E1020–E1029. doi: 10.1152/ajpendo.00637.2013 CrossRefPubMedGoogle Scholar
  14. Meex RC, Schrauwen P, Hesselink MK (2009) Modulation of myocellular fat stores: lipid droplet dynamics in health and disease. Am J Physiol Regul Integr Comp Physiol 297:R913–R924. doi: 10.1152/ajpregu.91053.2008 CrossRefPubMedGoogle Scholar
  15. Moro C, Bajpeyi S, Smith SR (2008) Determinants of intramyocellular triglyceride turnover: implications for insulin sensitivity. Am J Physiol Endocrinol Metab 294:E203–E213. doi: 10.1152/ajpendo.00624.2007 CrossRefPubMedGoogle Scholar
  16. Pette D, Staron RS (2000) Myosin isoforms, muscle fiber types, and transitions. Microsc Res Tech 50:500–509. doi: 10.1002/1097-0029(20000915) CrossRefPubMedGoogle Scholar
  17. Rakus D, Gizak A, Deshmukh A, Wisniewski JR (2015) Absolute quantitative profiling of the key metabolic pathways in slow and fast skeletal muscle. J Proteome Res 14:1400–1411. doi: 10.1021/pr5010357 CrossRefPubMedGoogle Scholar
  18. Rinnankoski-Tuikka R, Hulmi JJ, Torvinen S, Silvennoinen M, Lehti M, Kivela R, Reunanen H, Kujala UM, Kainulainen H (2014) Lipid droplet-associated proteins in high-fat fed mice with the effects of voluntary running and diet change. Metabolism 63:1031–1040. doi: 10.1016/j.metabol.2014.05.010 CrossRefPubMedGoogle Scholar
  19. Sacchetti M, Saltin B, Osada T, van Hall G (2002) Intramuscular fatty acid metabolism in contracting and non-contracting human skeletal muscle. J Physiol 540:387–395CrossRefPubMedPubMedCentralGoogle Scholar
  20. Sawano S, Komiya Y, Ichitsubo R, Ohkawa Y, Nakamura M, Tatsumi R, Ikeuchi Y, Mizunoya W (2016) A one-step immunostaining method to visualize rodent muscle fiber type within a single specimen. PLoS ONE 11:e0166080. doi: 10.1371/journal.pone.0166080 CrossRefPubMedPubMedCentralGoogle Scholar
  21. Schiaffino S (2010) Fibre types in skeletal muscle: a personal account. Acta Physiol (Oxf) 199:451–463. doi: 10.1111/j.1748-1716.2010.02130.x CrossRefGoogle Scholar
  22. Schiaffino S, Reggiani C (2011) Fiber types in mammalian skeletal muscles. Physiol Rev 91:1447–1531. doi: 10.1152/physrev.00031.2010 CrossRefPubMedGoogle Scholar
  23. Schiaffino S, Hanzlikova V, Pierobon S (1970) Relations between structure and function in rat skeletal muscle fibers. J Cell Biol 47:107–119CrossRefPubMedPubMedCentralGoogle Scholar
  24. Shaw CS, Jones DA, Wagenmakers AJ (2008) Network distribution of mitochondria and lipid droplets in human muscle fibres. Histochem Cell Biol 129:65–72. doi: 10.1007/s00418-007-0349-8 CrossRefPubMedGoogle Scholar
  25. Spangenburg EE, Pratt SJ, Wohlers LM, Lovering RM (2011) Use of BODIPY (493/503) to visualize intramuscular lipid droplets in skeletal muscle. J Biomed Biotechnol 2011:598358. doi: 10.1155/2011/598358 CrossRefPubMedPubMedCentralGoogle Scholar
  26. Suzuki M, Shinohara Y, Ohsaki Y, Fujimoto T (2011) Lipid droplets: size matters. J Electron Microsc (Tokyo) 60(Suppl 1):S101–116. doi: 10.1093/jmicro/dfr016 CrossRefGoogle Scholar
  27. Takahashi Y, Shinoda A, Furuya N, Harada E, Arimura N, Ichi I, Fujiwara Y, Inoue J, Sato R (2013) Perilipin-mediated lipid droplet formation in adipocytes promotes sterol regulatory element-binding protein-1 processing and triacylglyceride accumulation. PLoS ONE 8:e64605. doi: 10.1371/journal.pone.0064605 CrossRefPubMedPubMedCentralGoogle Scholar
  28. Tansey JT, Sztalryd C, Gruia-Gray J, Roush DL, Zee JV, Gavrilova O, Reitman ML, Deng CX, Li C, Kimmel AR, Londos C (2001) Perilipin ablation results in a lean mouse with aberrant adipocyte lipolysis, enhanced leptin production, and resistance to diet-induced obesity. Proc Natl Acad Sci U S A 98:6494–6499. doi: 10.1073/pnas.101042998 CrossRefPubMedPubMedCentralGoogle Scholar
  29. van Loon LJ (2004) Intramyocellular triacylglycerol as a substrate source during exercise. Proc Nutr Soc 63:301–307. doi: 10.1079/PNS2004347 CrossRefPubMedGoogle Scholar
  30. van Loon LJ, Goodpaster BH (2006) Increased intramuscular lipid storage in the insulin-resistant and endurance-trained state. Pflugers Arch 451:606–616. doi: 10.1007/s00424-005-1509-0 CrossRefPubMedGoogle Scholar
  31. van Loon LJ, Koopman R, Stegen JH, Wagenmakers AJ, Keizer HA, Saris WH (2003) Intramyocellular lipids form an important substrate source during moderate intensity exercise in endurance-trained males in a fasted state. J Physiol 553:611–625. doi: 10.1113/jphysiol.2003.052431 CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2017

Authors and Affiliations

  1. 1.Department of Bioresource Sciences, Faculty of AgricultureKyushu UniversityFukuokaJapan
  2. 2.Department of Animal Science, School of Veterinary MedicineKitasato UniversityTowadaJapan
  3. 3.Department of Food NutritionFukuoka Women’s Junior CollegeDazaifuJapan

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