Advertisement

Molecular and Cellular Biochemistry

, Volume 452, Issue 1–2, pp 29–39 | Cite as

The utrophin–beta 2 syntrophin complex regulates adipocyte lipid droplet size independent of adipogenesis

  • Sabrina Krautbauer
  • Markus Neumeier
  • Elisabeth M. Haberl
  • Rebekka Pohl
  • Susanne Feder
  • Kristina Eisinger
  • Lisa Rein-Fischboeck
  • Christa BuechlerEmail author
Article
  • 101 Downloads

Abstract

Utrophin is a widely expressed cytoskeleton protein and is associated with lipid droplets (LDs) in adipocytes. The scaffold protein beta 2 syntrophin (SNTB2) controls signaling events by recruiting distinct membrane and cytoskeletal proteins, and binds to utrophin. Here we show that SNTB2 forms a complex with utrophin in adipocytes. SNTB2 protein is strongly diminished when utrophin is low. Of note, knock-down of utrophin or SNTB2 enhances LD growth during adipogenesis. SNTB2 reduction has no effect on basal and induced lipolysis, and insulin-stimulated phosphorylation of Akt is normal. The antilipolytic activity of insulin is enhanced in adipocytes with low SNTB2, while knock-down of utrophin has no effect. Uptake of exogenously supplied oleate and linoleate is comparable in scrambled and SNTB2 siRNA-treated cells. In the fibroblasts, diminished SNTB2 is associated with lower proliferation. CCAAT/enhancer-binding protein alpha and sterol regulatory element-binding proteins which are critical transcription factors for adipogenesis are normally expressed. Consequently, maturation of cells with SNTB2 knock-down is not grossly impaired. In fibroblasts, SNTB2 is localized to filamentous and vesicular structures which are distinct from beta actin, alpha tubulin, endoplasmic reticulum, early endosomes, lysosomes and mitochondria. Collectively, our data provide evidence that the utrophin–SNTB2 complex regulates LD size without affecting adipogenesis.

Keywords

Adipogenesis Lipolysis Triglycerides Insulin 

Notes

Acknowledgements

The authors thank Prof. Marvin Adams for providing syntrophin-specific antibodies. The study was supported by a Grant from the German Research Foundation (BU 1141/8-1).

Compliance with ethical standards

Conflict of interest

The authors have no conflicts of interest to declare.

Supplementary material

11010_2018_3409_MOESM1_ESM.docx (31 kb)
Supplementary material 1 (DOCX 30 KB)

References

  1. 1.
    Buechler C, Krautbauer S, Eisinger K (2015) Adipose tissue fibrosis. World J Diabetes 6:548–553.  https://doi.org/10.4239/wjd.v6.i4.548 CrossRefGoogle Scholar
  2. 2.
    Mathieu PS, Loboa EG (2012) Cytoskeletal and focal adhesion influences on mesenchymal stem cell shape, mechanical properties, and differentiation down osteogenic, adipogenic, and chondrogenic pathways. Tissue Eng B 18:436–444.  https://doi.org/10.1089/ten.TEB.2012.0014 CrossRefGoogle Scholar
  3. 3.
    Sun K, Tordjman J, Clement K, Scherer PE (2013) Fibrosis and adipose tissue dysfunction. Cell Metab 18:470–477.  https://doi.org/10.1016/j.cmet.2013.06.016 CrossRefGoogle Scholar
  4. 4.
    Bhat HF, Adams ME, Khanday FA (2013) Syntrophin proteins as Santa Claus: role(s) in cell signal transduction. Cell Mol Life Sci 70:2533–2554.  https://doi.org/10.1007/s00018-012-1233-9 CrossRefGoogle Scholar
  5. 5.
    Iwata Y, Sampaolesi M, Shigekawa M, Wakabayashi S (2004) Syntrophin is an actin-binding protein the cellular localization of which is regulated through cytoskeletal reorganization in skeletal muscle cells. Eur J Cell Biol 83:555–565CrossRefGoogle Scholar
  6. 6.
    Squire S, Raymackers JM, Vandebrouck C, Potter A, Tinsley J, Fisher R, Gillis JM, Davies KE (2002) Prevention of pathology in mdx mice by expression of utrophin: analysis using an inducible transgenic expression system. Hum Mol Genet 11:3333–3344CrossRefGoogle Scholar
  7. 7.
    Sheng R, Chen Y, Yung Gee H, Stec E, Melowic HR, Blatner NR, Tun MP, Kim Y, Kallberg M, Fujiwara TK, Hye Hong J, Pyo Kim K, Lu H, Kusumi A, Goo Lee M, Cho W (2012) Cholesterol modulates cell signaling and protein networking by specifically interacting with PDZ domain-containing scaffold proteins. Nat Commun 3:1249.  https://doi.org/10.1038/ncomms2221 CrossRefGoogle Scholar
  8. 8.
    Buechler C, Bauer S (2012) ATP binding cassette transporter A1 (ABCA1) associated proteins: potential drug targets in the metabolic syndrome and atherosclerotic disease? Curr Pharm Biotechnol 13:319–330CrossRefGoogle Scholar
  9. 9.
    Ort T, Voronov S, Guo J, Zawalich K, Froehner SC, Zawalich W, Solimena M (2001) Dephosphorylation of beta2-syntrophin and Ca2+/mu-calpain-mediated cleavage of ICA512 upon stimulation of insulin secretion. Embo J 20:4013–4023CrossRefGoogle Scholar
  10. 10.
    Hebel T, Eisinger K, Neumeier M, Rein-Fischboeck L, Pohl R, Meier EM, Boettcher A, Froehner SC, Adams ME, Liebisch G, Krautbauer S, Buechler C (2015) Lipid abnormalities in alpha/beta2-syntrophin null mice are independent from ABCA1. Biochim Biophys Acta 1851:527–536.  https://doi.org/10.1016/j.bbalip.2015.01.012 CrossRefGoogle Scholar
  11. 11.
    Romo-Yanez J, Montanez C, Salazar-Olivo LA (2011) Dystrophins and DAPs are expressed in adipose tissue and are regulated by adipogenesis and extracellular matrix. Biochem Biophys Res Commun 404:717–722.  https://doi.org/10.1016/j.bbrc.2010.12.049 CrossRefGoogle Scholar
  12. 12.
    Haenggi T, Schaub MC, Fritschy JM (2005) Molecular heterogeneity of the dystrophin-associated protein complex in the mouse kidney nephron: differential alterations in the absence of utrophin and dystrophin. Cell Tissue Res 319:299–313.  https://doi.org/10.1007/s00441-004-0999-y CrossRefGoogle Scholar
  13. 13.
    Ding Y, Wu Y, Zeng R, Liao K (2012) Proteomic profiling of lipid droplet-associated proteins in primary adipocytes of normal and obese mouse. Acta Biochim Biophys Sin 44:394–406.  https://doi.org/10.1093/abbs/gms008 CrossRefGoogle Scholar
  14. 14.
    Eisinger K, Rein-Fischboeck L, Pohl R, Meier EM, Krautbauer S, Buechler C (2016) The adaptor protein alpha-syntrophin regulates adipocyte lipid droplet growth. Exp Cell Res 345:100–107.  https://doi.org/10.1016/j.yexcr.2016.05.020 CrossRefGoogle Scholar
  15. 15.
    Krautbauer S, Eisinger K, Hader Y, Buechler C (2014) Free fatty acids and IL-6 induce adipocyte galectin-3 which is increased in white and brown adipose tissues of obese mice. Cytokine 69:263–271.  https://doi.org/10.1016/j.cyto.2014.06.016 CrossRefGoogle Scholar
  16. 16.
    Krautbauer S, Eisinger K, Neumeier M, Hader Y, Buettner R, Schmid PM, Aslanidis C, Buechler C (2014) Free fatty acids, lipopolysaccharide and IL-1alpha induce adipocyte manganese superoxide dismutase which is increased in visceral adipose tissues of obese rodents. PLoS ONE 9:e86866.  https://doi.org/10.1371/journal.pone.0086866 CrossRefGoogle Scholar
  17. 17.
    Bauer S, Wanninger J, Schmidhofer S, Weigert J, Neumeier M, Dorn C, Hellerbrand C, Zimara N, Schaffler A, Aslanidis C, Buechler C (2011) Sterol regulatory element-binding protein 2 (SREBP2) activation after excess triglyceride storage induces chemerin in hypertrophic adipocytes. Endocrinology 152:26–35.  https://doi.org/10.1210/en.2010-1157 CrossRefGoogle Scholar
  18. 18.
    Aitchison AJ, Arsenault DJ, Ridgway ND (2015) Nuclear-localized CTP:phosphocholine cytidylyltransferase alpha regulates phosphatidylcholine synthesis required for lipid droplet biogenesis. Mol Biol Cell 26:2927–2938.  https://doi.org/10.1091/mbc.E15-03-0159 CrossRefGoogle Scholar
  19. 19.
    Keller P, Petrie JT, De Rose P, Gerin I, Wright WS, Chiang SH, Nielsen AR, Fischer CP, Pedersen BK, MacDougald OA (2008) Fat-specific protein 27 regulates storage of triacylglycerol. J Biol Chem 283:14355–14365.  https://doi.org/10.1074/jbc.M708323200 CrossRefGoogle Scholar
  20. 20.
    Qi Y, Sun L, Yang H (2017) Lipid droplet growth and adipocyte development: mechanistically distinct processes connected by phospholipids. Biochim Biophys Acta 1862:1273–1283.  https://doi.org/10.1016/j.bbalip.2017.06.016 CrossRefGoogle Scholar
  21. 21.
    Dalen KT, Schoonjans K, Ulven SM, Weedon-Fekjaer MS, Bentzen TG, Koutnikova H, Auwerx J, Nebb HI (2004) Adipose tissue expression of the lipid droplet-associating proteins S3-12 and perilipin is controlled by peroxisome proliferator-activated receptor-gamma. Diabetes 53:1243–1252CrossRefGoogle Scholar
  22. 22.
    Greenberg AS, Coleman RA, Kraemer FB, McManaman JL, Obin MS, Puri V, Yan QW, Miyoshi H, Mashek DG (2011) The role of lipid droplets in metabolic disease in rodents and humans. J Clin Invest 121:2102–2110.  https://doi.org/10.1172/JCI46069 CrossRefGoogle Scholar
  23. 23.
    Ducharme NA, Bickel PE (2008) Lipid droplets in lipogenesis and lipolysis. Endocrinology 149:942–949.  https://doi.org/10.1210/en.2007-1713 CrossRefGoogle Scholar
  24. 24.
    Fruhbeck G, Mendez-Gimenez L, Fernandez-Formoso JA, Fernandez S, Rodriguez A (2014) Regulation of adipocyte lipolysis. Nutr Res Rev 27:63–93.  https://doi.org/10.1017/S095442241400002X CrossRefGoogle Scholar
  25. 25.
    Krautbauer S, Eisinger K, Hader Y, Neumeier M, Buechler C (2014) Manganese superoxide dismutase knock-down in 3T3-L1 preadipocytes impairs subsequent adipogenesis. Mol Cell Biochem 393:69–76.  https://doi.org/10.1007/s11010-014-2047-x CrossRefGoogle Scholar
  26. 26.
    Ariotti N, Murphy S, Hamilton NA, Wu L, Green K, Schieber NL, Li P, Martin S, Parton RG (2012) Postlipolytic insulin-dependent remodeling of micro lipid droplets in adipocytes. Mol Biol Cell 23:1826–1837.  https://doi.org/10.1091/mbc.E11-10-0847 CrossRefGoogle Scholar
  27. 27.
    Ding S, Mersmann HJ (2001) Fatty acids modulate porcine adipocyte differentiation and transcripts for transcription factors and adipocyte-characteristic proteins* J Nutr Biochem 12:101–108CrossRefGoogle Scholar
  28. 28.
    Ouyang D, Ye Y, Guo D, Yu X, Chen J, Qi J, Tan X, Zhang Y, Ma Y, Li Y (2015) MicroRNA-125b-5p inhibits proliferation and promotes adipogenic differentiation in 3T3-L1 preadipocytes. Acta Biochim Biophys Sin 47:355–361.  https://doi.org/10.1093/abbs/gmv024 CrossRefGoogle Scholar
  29. 29.
    Tang HN, Man XF, Liu YQ, Guo Y, Tang AG, Liao EY, Zhou HD (2015) Dose-dependent effects of neuropeptide Y on the regulation of preadipocyte proliferation and adipocyte lipid synthesis via the PPARgamma pathways. Endocr J 62:835–846.  https://doi.org/10.1507/endocrj.EJ15-0133 CrossRefGoogle Scholar
  30. 30.
    Farmer SR (2006) Transcriptional control of adipocyte formation. Cell Metab 4:263–273.  https://doi.org/10.1016/j.cmet.2006.07.001 CrossRefGoogle Scholar
  31. 31.
    Rybakova IN, Patel JR, Davies KE, Yurchenco PD, Ervasti JM (2002) Utrophin binds laterally along actin filaments and can couple costameric actin with sarcolemma when overexpressed in dystrophin-deficient muscle. Mol Biol Cell 13:1512–1521.  https://doi.org/10.1091/mbc.01-09-0446 CrossRefGoogle Scholar
  32. 32.
    Tan JS, Seow CJ, Goh VJ, Silver DL (2014) Recent advances in understanding proteins involved in lipid droplet formation, growth and fusion. J Genet Genomics 41:251–259.  https://doi.org/10.1016/j.jgg.2014.03.003 CrossRefGoogle Scholar
  33. 33.
    Xu L, Zhou L, Li P (2012) CIDE proteins and lipid metabolism. Arterioscler Thromb Vasc Biol 32:1094–1098.  https://doi.org/10.1161/ATVBAHA.111.241489 CrossRefGoogle Scholar
  34. 34.
    Bostrom P, Rutberg M, Ericsson J, Holmdahl P, Andersson L, Frohman MA, Boren J, Olofsson SO (2005) Cytosolic lipid droplets increase in size by microtubule-dependent complex formation. Arterioscler Thromb Vasc Biol 25:1945–1951.  https://doi.org/10.1161/01.ATV.0000179676.41064.d4 CrossRefGoogle Scholar
  35. 35.
    Boschi F, Rizzatti V, Zamboni M, Sbarbati A (2015) Models of lipid droplets growth and fission in adipocyte cells. Exp Cell Res 336:253–262.  https://doi.org/10.1016/j.yexcr.2015.06.001 CrossRefGoogle Scholar
  36. 36.
    Capurso C, Capurso A (2012) From excess adiposity to insulin resistance: the role of free fatty acids. Vasc Pharmacol 57:91–97.  https://doi.org/10.1016/j.vph.2012.05.003 CrossRefGoogle Scholar
  37. 37.
    Jo J, Gavrilova O, Pack S, Jou W, Mullen S, Sumner AE, Cushman SW, Periwal V (2009) Hypertrophy and/or hyperplasia: dynamics of adipose tissue growth. PLoS Comput Biol 5:e1000324.  https://doi.org/10.1371/journal.pcbi.1000324 CrossRefGoogle Scholar
  38. 38.
    Wueest S, Rapold RA, Rytka JM, Schoenle EJ, Konrad D (2009) Basal lipolysis, not the degree of insulin resistance, differentiates large from small isolated adipocytes in high-fat fed mice. Diabetologia 52:541–546.  https://doi.org/10.1007/s00125-008-1223-5 CrossRefGoogle Scholar
  39. 39.
    Qi Y, Kapterian TS, Du X, Ma Q, Fei W, Zhang Y, Huang X, Dawes IW, Yang H (2016) CDP-diacylglycerol synthases regulate the growth of lipid droplets and adipocyte development. J Lipid Res 57:767–780.  https://doi.org/10.1194/jlr.M060574 CrossRefGoogle Scholar
  40. 40.
    Belkin AM, Burridge K (1995) Localization of utrophin and aciculin at sites of cell-matrix and cell-cell adhesion in cultured cells. Exp Cell Res 221:132–140.  https://doi.org/10.1006/excr.1995.1360 CrossRefGoogle Scholar
  41. 41.
    Eisinger K, Rein-Fischboeck L, Neumeier M, Schmidhofer S, Pohl R, Haberl EM, Liebisch G, Kopp A, Schmid A, Krautbauer S, Buechler C (2018) Alpha-syntrophin deficient mice are protected from adipocyte hypertrophy and ectopic triglyceride deposition in obesity. Exp Mol Pathol 104:212–221.  https://doi.org/10.1016/j.yexmp.2018.04.003 CrossRefGoogle Scholar
  42. 42.
    Elizalde M, Ryden M, van Harmelen V, Eneroth P, Gyllenhammar H, Holm C, Ramel S, Olund A, Arner P, Andersson K (2000) Expression of nitric oxide synthases in subcutaneous adipose tissue of nonobese and obese humans. J Lipid Res 41:1244–1251Google Scholar
  43. 43.
    Verkman AS, Anderson MO, Papadopoulos MC (2014) Aquaporins: important but elusive drug targets. Nat Rev Drug Discov 13:259–277.  https://doi.org/10.1038/nrd4226 CrossRefGoogle Scholar
  44. 44.
    Buechler C, Boettcher A, Bared SM, Probst MC, Schmitz G (2002) The carboxyterminus of the ATP-binding cassette transporter A1 interacts with a beta2-syntrophin/utrophin complex. Biochem Biophys Res Commun 293:759–765.  https://doi.org/10.1016/S0006-291X(02)00303-0 CrossRefGoogle Scholar
  45. 45.
    Tamehiro N, Park MH, Hawxhurst V, Nagpal K, Adams ME, Zannis VI, Golenbock DT, Fitzgerald ML (2015) LXR agonism up-regulates the macrophage ABCA1/syntrophin protein complex which can bind apoA-I and stabilized ABCA1 protein, but complex loss does not inhibit lipid efflux. Biochemistry 54:6931–6941.  https://doi.org/10.1021/acs.biochem.5b00894 CrossRefGoogle Scholar
  46. 46.
    Lyssand JS, Lee KS, DeFino M, Adams ME, Hague C (2011) Syntrophin isoforms play specific functional roles in the alpha1D-adrenergic receptor/DAPC signalosome. Biochem Biophys Res Commun 412:596–601.  https://doi.org/10.1016/j.bbrc.2011.08.004 CrossRefGoogle Scholar
  47. 47.
    Neumeier M, Krautbauer S, Schmidhofer S, Hader Y, Eisinger K, Eggenhofer E, Froehner SC, Adams ME, Mages W, Buechler C (2013) Adiponectin receptor 1 C-terminus interacts with PDZ-domain proteins such as syntrophins. Exp Mol Pathol 95:180–186.  https://doi.org/10.1016/j.yexmp.2013.07.002 CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Internal Medicine IUniversity Hospital of RegensburgRegensburgGermany

Personalised recommendations