Histochemistry and Cell Biology

, Volume 131, Issue 5, pp 575–581 | Cite as

Adipophilin distribution and colocalisation with lipid droplets in skeletal muscle

  • Christopher S. ShawEmail author
  • Mark Sherlock
  • Paul M. Stewart
  • Anton J. M. Wagenmakers
Original Paper


Intramyocellular lipids (IMCL) are stored as discrete lipid droplets which are associated with a number of proteins. The lipid droplet-associated protein adipophilin (the human orthologue of adipose differentiation-related protein) is ubiquitously expressed and is one of the predominant lipid droplet-proteins in skeletal muscle. The aim of this study was to investigate the subcellular distribution of adipophilin in human muscle fibres and to measure the colocalisation of adipophilin with IMCL. Muscle biopsies from six lean male cyclists (BMI 23.4 ± 0.4, aged 31 ± 2 years, W max 346 ± 8) were stained for myosin heavy chain type 1, IMCL, adipophilin and mitochondria using immunofluorescence and viewed with widefield and confocal fluorescence microscopy. The present study shows that like IMCL, the adipophilin content is ~twofold greater in type I skeletal muscle fibres and is situated in the areas between the mitochondrial network. Colocalisation analysis demonstrated that 61 ± 2% of IMCL contain adipophilin. Although the majority of adipophilin is contained within IMCL, 36 ± 4% of adipophilin is not associated with IMCL. In conclusion, this study indicates that the IMCL pool is heterogenous, as the majority but not all IMCL contain adipophilin.


Adipose-differentiation related protein (ADRP) Intramyocellular lipids (IMCL) PAT-proteins Fluorescence microscopy Skeletal muscle 



The study was performed at the Wellcome Trust Clinical Research Facility, Queen Elizabeth Hospital, and we thank Heather Jones and Joanna Finney for nursing support during the study. The antibody against myosin (human slow fibres, A4.840) used in the study was developed by Dr. Blau and was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by The University of Iowa, Department of Biological Sciences, Iowa City, IA 52242.


  1. Bartz R, Zehmer JK, Zhu M, Chen Y, Serrero G, Zhao Y, Liu P (2007) Dynamic activity of lipid droplets: protein phosphorylation and GTP-mediated protein translocation. J Proteome Res 6:3256–3265PubMedCrossRefGoogle Scholar
  2. Bays H, Mandarino L, DeFronzo RA (2004) Mechanisms of endocrine disease. Role of the adipocyte, free fatty acids, and ectopic fat in pathogenesis of type 2 diabetes mellitus: peroxisomal proliferator-activates receptor agonists provide a rational therapeutic approach. J Clin Endocrinol Metab 89:463–478PubMedCrossRefGoogle Scholar
  3. Bell M, Wang H, Chen H, McLenithan JC, Gong DW, Yang RZ et al (2008) Consequences of lipid droplet coat proteins down-regulation in liver cells: abnormal lipid droplet metabolism and induction of insulin resistance. Diabetes 57:2037–2045PubMedCrossRefGoogle Scholar
  4. Bergstrom J (1975) Percutaneous needle biopsy of skeletal muscle in physiological and clinical research. Scand J Clin Lab Invest 35:609–616PubMedCrossRefGoogle Scholar
  5. Brasaemle DL, Barber T, Wolins NE, Serrero G, Blanchette-Mackie EJ, Londos C (1997) Adipose differentiation-related protein is an ubiquitously expressed lipid storage droplet-associated protein. J Lipid Res 38:2249–2263PubMedGoogle Scholar
  6. Brasaemle DL, Dolios G, Shapiro L, Wang R (2004) Proteomic analysis of proteins associated with lipid droplets of basal and lipolytically stimulated 3T3-L1 adipocytes. J Biol Chem 279:46835–46842PubMedCrossRefGoogle Scholar
  7. Ducharme NA, Bickel PE (2008) Lipid droplets in lipogenesis and lipolysis. Endocrinology 149:942–949PubMedCrossRefGoogle Scholar
  8. Frayn KN, Arner P, Yki-Järvinen H (2006) Fatty acid metabolism in adipose tissue, muscle and liver in health and disease. Essays Biochem 42:89–103PubMedCrossRefGoogle Scholar
  9. Fujimoto T, Ohsaki Y, Cheng J, Suzuki M, Shinohara (2008) Lipid droplets: a classic organelle with new outfits. Histochem Cell Biol 130:263–279PubMedCrossRefGoogle Scholar
  10. Goodman JM (2008) The gregarious lipid droplet. J Biol Chem 283:28005–28009PubMedCrossRefGoogle Scholar
  11. Guilherme A, Virbasius JV, Puri V, Czech MP (2008) Adipocyte dysfunctions linking obesity to insulin resistance and type 2 diabetes. Nat Rev Mol Cell Biol 9:367–377PubMedCrossRefGoogle Scholar
  12. Howald H, Hoppeler H, Claassen H, Mathieu O, Straub R (1985) Influences of endurance training on the ultrastructural composition of the different muscle fiber types in humans. Pflugers Arch 403:369–376PubMedCrossRefGoogle Scholar
  13. Imamura M, Inoguchi T, Ikuyama S, Taniguchi S, Kobayashi K, Nakashima N, Nawata H (2002) ADRP stimulates lipid accumulation and lipid droplet formation in murine fibroblasts. Am J Physiol Endocrinol Metab 283:E775–E783PubMedGoogle Scholar
  14. Jocken JW, Smit E, Goossens GH, Essers YP, van Baak MA, Mensink M, Saris WH, Blaak EE (2008) Adipose triglyceride lipase (ATGL) expression in human skeletal muscle is type I (oxidative) fiber specific. Histochem Cell Biol 129:535–538PubMedCrossRefGoogle Scholar
  15. Kayar SR, Hoppeler H, Essen-Gustavsson B, Schwerzmann K (1988) The similarity of mitochondrial distribution in equine skeletal muscles of differing oxidative capacity. J Exp Biol 137:253–263PubMedGoogle Scholar
  16. Koopman R, Schaart G, Hesselink MKC (2001) Optimisation of oil red O staining permits combination with immunofluorescence and automated quantification of lipids. Histochem Cell Biol 116:63–68PubMedGoogle Scholar
  17. Koopman R, Manders RJF, Jonkers RAM, Hul GBJ, Kuipers H, van Loon LJC (2005) Intramyocellular lipid and glycogen content are reduced following resistance exercise in untrained healthy males. Eur J Appl Physiol 96:525–534PubMedCrossRefGoogle Scholar
  18. Langfort J, Ploug T, Ihlemann J, Saldo M, Holm C, Galbo H (1999) Expression of hormone-sensitive lipase and its regulation by adrenaline in skeletal muscle. Biochem J 340:459–465PubMedCrossRefGoogle Scholar
  19. Lewis GF, Carpentier A, Adeli K, Giacca A (2002) Disordered fat storage and mobilization in the pathogenesis of insulin resistance and type 2 diabetes. Endocr Rev 23:201–229PubMedCrossRefGoogle Scholar
  20. Listenberger LL, Ostemeyer-Fay AG, Goldberg EB, Brown WJ, Brown DA (2007) Adipocyte differentiation-related protein reduces the lipid droplet association of adipose triglyceride lipase and slows triacylglycerol turnover. J Lipid Res 48:2751–2761PubMedCrossRefGoogle Scholar
  21. Magnusson B, Asp L, Boström P, Ruiz M, Stillemark-Billton P, Lindén D, Borén J, Olofsson SO (2006) Adipocyte differentiation-related protein promotes fatty acid storage in cytosolic triglycerides and inhibits secretion of very low-density lipoproteins. Arterioscler Thromb Vasc Biol 26:1566–1571PubMedCrossRefGoogle Scholar
  22. Malenfant P, Joanisse DR, Theriault R, Goodpaster BH, Kelley DE, Simoneau JA (2001) Fat content in individual muscle fibers of lean and obese subjects. Int J Obes Relat Metab Disord 25:1316–1321PubMedCrossRefGoogle Scholar
  23. Phillips SA, Choe CC, Ciaraldi TP, Greenberg AS, Kong AP, Baxi SC, Christiansen L, Mudaliar SR, Henry RR (2005) Adipocyte differentiation-related protein in human skeletal muscle: relationship to insulin sensitivity. Obesity Res 13:1321–1329CrossRefGoogle Scholar
  24. Prats C, Donsmark M, Qvortrup K, Londos C, Sztalryd C, Holm C, Galbo H, Ploug T (2006) Decrease in intramuscular lipid droplets and translocation of HSL in response to muscle contraction and epinephrine. J Lipid Res 47:2392–2399PubMedCrossRefGoogle Scholar
  25. Shaw CS, Jones DA, Wagenmakers AJM (2008) Network distribution of mitochondria and lipid droplets in human muscle fibres. Histochem Cell Biol 129:65–72PubMedCrossRefGoogle Scholar
  26. Shulman GI (2000) Cellular mechanisms of insulin resistance. J Clin Invest 106:171–176PubMedCrossRefGoogle Scholar
  27. Tarnopolsky MA, Rennie CD, Robertshaw HA, Fedak-Tarnopolsky SN, Devries MC, Hamadeh MJ (2006) The influence of endurance exercise training and sex on intramyocellular lipid and mitochondrial ultrastructure. Am J Physiol Regul Integr Comp Physiol 292:R1271–R1278PubMedGoogle Scholar
  28. Thiele C, Spandl J (2008) Cell biology of lipid droplets. Curr Opin Cell Biol 20:1–8CrossRefGoogle Scholar
  29. van Loon LJC (2004) Use of intramuscular triacylglycerol as a substrate source during exercise in humans. J Appl Physiol 97:1170–1187PubMedCrossRefGoogle Scholar
  30. van Loon LJC, Koopman R, Stegen JHCH, Wagenmakers AJM, Keizer HA, Saris WHM (2003) Intramyocellular lipids form an important substrate source during moderate intensity exercise in endurance-trained males in a fasted state. J Physiol 553:611–625PubMedCrossRefGoogle Scholar
  31. van Loon LJC, Koopman R, Manders R, van der Weegen W, van Kranenburg GP, Keizer HA (2004) Intramyocellular lipid content in type 2 diabetes patients compared with overweight sedentary men and highly trained endurance athletes. Am J Physiol Endocrinol Metab 287:E558–E565PubMedCrossRefGoogle Scholar
  32. Wolins NE, Quaynor BK, Skinner JR, Schoenfish MJ, Tzekov A, Bickel PE (2005) S3-12, adipophilin, and TIP47 package lipid in adipocytes. J Biol Chem 280:19146–19155PubMedCrossRefGoogle Scholar
  33. Wolins NE, Brasaemle DL, Bickel PE (2006) A proposed model of fat packaging by exchangeable lipid droplet proteins. FEBS Lett 580:5484–5491PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Christopher S. Shaw
    • 1
    Email author
  • Mark Sherlock
    • 2
  • Paul M. Stewart
    • 2
  • Anton J. M. Wagenmakers
    • 1
  1. 1.Exercise Metabolism Research Group, School of Sport and Exercise SciencesUniversity of BirminghamBirminghamUK
  2. 2.School of Clinical and Experimental Medicine, College of Medicine and Dental SciencesUniversity of BirminghamBirminghamUK

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