Abstract
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.
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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 6:3256–3265
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 89:463–478
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 57:2037–2045
Bergstrom J (1975) Percutaneous needle biopsy of skeletal muscle in physiological and clinical research. Scand J Clin Lab Invest 35:609–616
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 38:2249–2263
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 279:46835–46842
Ducharme NA, Bickel PE (2008) Lipid droplets in lipogenesis and lipolysis. Endocrinology 149:942–949
Frayn KN, Arner P, Yki-Järvinen H (2006) Fatty acid metabolism in adipose tissue, muscle and liver in health and disease. Essays Biochem 42:89–103
Fujimoto T, Ohsaki Y, Cheng J, Suzuki M, Shinohara (2008) Lipid droplets: a classic organelle with new outfits. Histochem Cell Biol 130:263–279
Goodman JM (2008) The gregarious lipid droplet. J Biol Chem 283:28005–28009
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 9:367–377
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 403:369–376
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–E783
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 129:535–538
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 137:253–263
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 116:63–68
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 96:525–534
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 340:459–465
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 23:201–229
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 48:2751–2761
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 26:1566–1571
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 25:1316–1321
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 13:1321–1329
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 47:2392–2399
Shaw CS, Jones DA, Wagenmakers AJM (2008) Network distribution of mitochondria and lipid droplets in human muscle fibres. Histochem Cell Biol 129:65–72
Shulman GI (2000) Cellular mechanisms of insulin resistance. J Clin Invest 106:171–176
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–R1278
Thiele C, Spandl J (2008) Cell biology of lipid droplets. Curr Opin Cell Biol 20:1–8
van Loon LJC (2004) Use of intramuscular triacylglycerol as a substrate source during exercise in humans. J Appl Physiol 97:1170–1187
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 553:611–625
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–E565
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 280:19146–19155
Wolins NE, Brasaemle DL, Bickel PE (2006) A proposed model of fat packaging by exchangeable lipid droplet proteins. FEBS Lett 580:5484–5491
Acknowledgments
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.
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Shaw, C.S., Sherlock, M., Stewart, P.M. et al. Adipophilin distribution and colocalisation with lipid droplets in skeletal muscle. Histochem Cell Biol 131, 575–581 (2009). https://doi.org/10.1007/s00418-009-0558-4
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DOI: https://doi.org/10.1007/s00418-009-0558-4