Advertisement

Histochemistry and Cell Biology

, Volume 130, Issue 1, pp 91–103 | Cite as

New aspects of obscurin in human striated muscles

  • Lena Carlsson
  • Ji-Guo Yu
  • Lars-Eric Thornell
Original Paper

Abstract

Obscurin is a giant protein (700–800 kDa) present in both skeletal muscles and myocardium. According to animal studies, obscurin interacts with myofibrillar Z-discs during early muscle development, but is translocalised to be predominantly associated with the M-bands in mature muscles. The proposed function for obscurin is in the assembly and organisation of myosin into regular A-bands during formation of new sarcomeres. In the present study, the precise localisation of obscurin in developing and mature normal human striated muscle is presented for the first time. We show that obscurin surrounded myofibrils at the M-band level in both developing and mature human skeletal and heart muscles, which is partly at variance with that observed in animals. At maturity, obscurin also formed links between the peripheral myofibrils and the sarcolemma, and was a distinct component of the neuromuscular junctions. Obscurin should therefore be regarded as an additional component of the extrasarcomeric cytoskeleton. To test this function of obscurin, biopsies from subjects with exercise-induced delayed onset muscle soreness (DOMS) were examined. In these subjects, myofibrillar alterations related to sarcomerogenesis are observed. Our immunohistochemical analysis revealed that obscurin was never lacking in myofibrillar alterations, but was either preserved at the M-band level or diffusely spread over the sarcomeres. As myosin was absent in such areas but later reincorporated in the newly formed sarcomeres, our results support that obscurin also might play an important role in the formation and maintenance of A-bands.

Keywords

Cytoskeleton Heart Skeletal muscle Development Sarcomerogenesis DOMS 

Notes

Acknowledgments

We wish to thank Mrs. Margaretha Enerstedt for technical assistance and Prof. M Gautel, Cardiovascular Division and Randall Division for Cell and Molecular Biophysics, New Hunt´s House, King’s College London, UK for the gift of obscurin antibodies. Supported by Grants from the Swedish Research Council (12x 3934), the Swedish National Centre for Research in Sports (98/04, 108/05, 109/06) and the Medical Faculty of Umeå University, Sweden.

References

  1. Agarkova I, Schoenauer R, Ehler E, Carlsson L, Carlsson E, Thornell LE, Perriard JC (2004) The molecular composition of the sarcomeric M-band correlates with muscle fiber type. Eur J Cell Biol 83:193–204PubMedCrossRefGoogle Scholar
  2. Arimura T, Matsumoto Y, Okazaki O, Hayashi T, Takahashi M, Inagaki N, Hinohara K, Ashizawa N, Yano K, Kimura A (2007) Structural analysis of obscurin gene in hypertrophic cardiomyopathy. Biochem Biophys Res Commun 362:281–287PubMedCrossRefGoogle Scholar
  3. Armani A, Galli S, Giacomello E, Bagnato P, Barone V, Rossi D, Sorrentino V (2006) Molecular interactions with obscurin are involved in the localization of muscle-specific small ankyrin1 isoforms to subcompartments of the sarcoplasmic reticulum. Exp Cell Res 312:3546–3558PubMedCrossRefGoogle Scholar
  4. Askanas V, Bornemann A, Engel WK (1990) Immunocytochemical localization of desmin at human neuromuscular junctions. Neurology 40:949–953PubMedGoogle Scholar
  5. Bagnato P, Barone V, Giacomello E, Rossi D, Sorrentino V (2003) Binding of an ankyrin-1 isoform to obscurin suggests a molecular link between the sarcoplasmic reticulum and myofibrils in striated muscles. J Cell Biol 160:245–253PubMedCrossRefGoogle Scholar
  6. Bang ML, Centner T, Fornoff F, Geach AJ, Gotthardt M, McNabb M, Witt CC, Labeit D, Gregorio CC, Granzier H, Labeit S (2001) The complete gene sequence of titin, expression of an unusual approximately 700-kDa titin isoform, and its interaction with obscurin identify a novel Zline to I-band linking system. Circ Res 89:1065–1072PubMedCrossRefGoogle Scholar
  7. Berthier C, Blaineau S (1997) Supramolecular organization of the subsarcolemmal cytoskeleton of adult skeletal muscle fibers. Biol Cell 89:413–434PubMedCrossRefGoogle Scholar
  8. Bonnemann CG, Laing NG (2004) Myopathies resulting from mutations in sarcomeric proteins. Curr Opin Neurol 17:529–537PubMedCrossRefGoogle Scholar
  9. Borisov AB, Raeker MO, Kontrogianni-Konstantopoulos A, Yang K, Kurnit DM, Bloch RJ, Russell MW (2003) Rapid response of cardiac obscurin gene cluster to aortic stenosis: differential activation of Rho-GEF and MLCK and involvement in hypertrophic growth. Biochem Biophys Res Commun 310:910–918PubMedCrossRefGoogle Scholar
  10. Borisov AB, Kontrogianni-Konstantopoulos A, Bloch RJ, Westfall MV, Russell MW (2004) Dynamics of obscurin localization during differentiation and remodeling of cardiac myocytes: obscurin as an integrator of myofibrillar structure. J Histochem Cytochem 52:1117–1127PubMedCrossRefGoogle Scholar
  11. Borisov AB, Sutter SB, Kontrogianni-Konstantopoulos A, Bloch RJ, Westfall MV, Russell MW (2006) Essential role of obscurin in cardiac myofibrillogenesis and hypertrophic response: evidence from small interfering RNA-mediated gene silencing. Histochem Cell Biol 123:227–238CrossRefGoogle Scholar
  12. Borisov AB, Raeker MÖ, Russell MW (2007) Developmental expression and differential cellular localization of obscurin and obscurin-associated kinase in cardiac muscle cells. J Cell Biochem (published on line)Google Scholar
  13. Borisov AB, Martynova MG, Russell MW (2008) Early incorporation of obscurin into nascent sarcomeres: implication for myofibrillar assembly during cardiac myogenesis. Histochem Cell Biol (published on line)Google Scholar
  14. Bowman AL, Kontrogianni-Konstantopoulos A, Hirsch SS, Geisler SB, Gonzalez-Serratos H, Russell MW, Bloch RJ (2007) Different obscurin isoforms localize to distinct sites at sarcomeres. FEBS Lett 581:1549–1554PubMedCrossRefGoogle Scholar
  15. Capetanaki Y, Bloch RJ, Kouloumenta A, Mavroidis M, Psarras S (2007) Muscle intermediate filaments and their links to membranes and membranous organelles. Exp Cell Res 313:2063–2076PubMedCrossRefGoogle Scholar
  16. Carlsson L, Thornell L-E (2001) Desmin-related myopathies in mice and man. Acta Physiol Scand 171:341–348PubMedCrossRefGoogle Scholar
  17. Carlsson L, Li Z, Paulin D, Thornell L-E (1999) Nestin is expressed during development and in myotendinous and neuromuscular junctions in wild type and desmin knock-out mice. Exp Cell Res 251:213–223PubMedCrossRefGoogle Scholar
  18. Carlsson L, Li Z, Paulin D, Price MG, Breckler J, Robson RM, Wiche G, Thornell L-E (2000) Differences in the distribution of synemin, paranemin and plectin in skeletal muscles of wild type and desmin knock-out mice. Histochem Cell Biol 114:39–47PubMedGoogle Scholar
  19. Carlsson L, Yu JG, Moza M, Carpen O, Thornell LE (2006) Myotilin—a prominent marker of myofibrillar remodelling. Neuromuscul Disord 17:61–68PubMedCrossRefGoogle Scholar
  20. Carpenter S, Karpati G (2001) Pathology of skeletal muscle. 2nd edn. Oxford University Press, New YorkGoogle Scholar
  21. Clark KA, McElhinny AS, Beckerle MC, Gregorio CC (2002) Striated muscle cytoarchitecture: an intricate web of form and function. Annu Rev Cell Dev Biol 18:637–706PubMedCrossRefGoogle Scholar
  22. Deshmukh L, Tyukhtenko S, Liu J, Fox JE, Qin J, Vinogradova O (2007) Structural insight into the interaction between platelet integrin alphaIIbbeta3 and cytoskeletal protein skelemin. J Biol Chem 282:32349–32356PubMedCrossRefGoogle Scholar
  23. Fridén J, Lieber RL (2001) Eccentric exercise-induced injuries to contractile and cytoskeletal muscle fibre components. Acta Physiol Scand 171:321–326PubMedCrossRefGoogle Scholar
  24. Fukuzawa A, Idowu S, Gautel M (2005) Complete human gene structure of obscurin: implications for isoform generation by differential splicing. J Muscle Res Cell Motil 26:427–434PubMedCrossRefGoogle Scholar
  25. Funatsu T, Kono E, Higuchi H, Kimura S, Ishiwata S, Yoshioka T, Maruyama K, Tsukita S (1993) Elastic filaments in situ in cardiac muscle: deep-etch replica analysis in combination with selective removal of actin and myosin filaments. J Cell Biol 120:711–724PubMedCrossRefGoogle Scholar
  26. Geisler SB, Robinson D, Hauringa M, Raeker MO, Borisov AB, Westfall MV, Russell MW (2007) Obscurin-like 1, OBSL1, is a novel cytoskeletal protein related to obscurin. Genomics 89:521–531PubMedCrossRefGoogle Scholar
  27. Granger BL, Lazarides E (1978) The existence of an insoluble Z disc scaffold in chicken skeletal muscle. Cell 15:1253–1268PubMedCrossRefGoogle Scholar
  28. Grove BK, Kurer V, Lehner C, Doetschman TC, Perriard JC, Eppenberger HM (1984) A new 185,000-dalton skeletal muscle protein detected by monoclonal antibodies. J Cell Biol 98:518–524PubMedCrossRefGoogle Scholar
  29. Hornemann T, Kempa S, Himmel M, Hayess K, Furst DO, Wallimann T (2003) Muscle-type creatine kinase interacts with central domains of the M-band proteins myomesin and M-protein. J Mol Biol 332:877–887PubMedCrossRefGoogle Scholar
  30. Kontrogianni-Konstantopoulos A, Jones EM, Van Rossum DB, Bloch RJ (2003) Obscurin is a ligand for small ankyrin 1 in skeletal muscle. Mol Biol Cell 14:1138–1148PubMedCrossRefGoogle Scholar
  31. Kontrogianni-Konstantopoulos A, Catino DH, Strong JC, Randall WR, Bloch RJ (2004) Obscurin regulates the organization of myosin into A bands. Am J Physiol Cell Physiol 287:C209–C217PubMedCrossRefGoogle Scholar
  32. Kontrogianni-Konstantopoulos A, Catino DH, Strong JC, Bloch RJ (2006a) De Novo Myofibrillogenesis in C2C12 Cells: Evidence for the Independent Assembly of M-lines and Z-disks. Am J Physiol Cell Physiol 290:C626–637PubMedCrossRefGoogle Scholar
  33. Kontrogianni-Konstantopoulos A, Bloch RJ (2006b) Obscurin: a multitasking muscle giant. J Muscle Res Cell Motil 26:419–426CrossRefGoogle Scholar
  34. Lange S, Agarkova I, Perriard JC, Ehler E (2005a) The sarcomeric M-band during development and in disease. J Muscle Res Cell Motil 26:375–379PubMedCrossRefGoogle Scholar
  35. Lange S, Xiang F, Yakovenko A, Vihola A, Hackman P, Rostkova E, Kristensen J, Brandmeier B, Franzen G, Hedberg B, Gunnarsson LG, Hughes SM, Marchand S, Sejersen T, Richard I, Edstrom L, Ehler E, Udd B, Gautel M (2005b) The kinase domain of titin controls muscle gene expression and protein turnover. Science 308:1599–1603PubMedCrossRefGoogle Scholar
  36. Lazarides E, Burridge K (1975) Alpha-actinin: immunofluorescent localization of a muscle structural protein in nonmuscle cells. Cell 6:289–298PubMedCrossRefGoogle Scholar
  37. Lieber RL, Shah S, Fridén J (2002) Cytoskeletal disruption after eccentric contraction-induced muscle injury. Clin Orthop:S90–S99Google Scholar
  38. Pierobon-Bormioli S (1981) Transverse sarcomere filamentous systems: “Z and M-cables.” J Muscle Res Cell Motil 2:401–413CrossRefGoogle Scholar
  39. Prasad V, Semwogerere D, Weeks ER (2007) Confocal microscopy of colloids. J Phys Condens Matter 19:1–25CrossRefGoogle Scholar
  40. Price MG (1984) Molecular analysis of intermediate filament cytoskeleton—putative load-bearing structure. Am J Physiol 246:H566–H572PubMedGoogle Scholar
  41. Price MG (1987) Skelemins: cytoskeletal proteins located at the periphery of M-discs in mammalian striated muscle. J Cell Biol 104:1325–1336PubMedCrossRefGoogle Scholar
  42. Price MG (1991) Striated muscle endosarcomeric and exosarcomeric lattices. In: Advances in structural biology. JAI Press Inc, Greenwich, pp 175–207Google Scholar
  43. Price MG, Gomer RH (1993) Skelemin, a cytoskeletal M-disc periphery protein, contains motifs of adhesion/recognition and intermediate filament proteins. J Biol Chem 268:21800–21810PubMedGoogle Scholar
  44. Raeker MO, Su F, Geisler SB, Borisov AB, Kontrogianni-Konstantopoulos A, Lyons SE, Russell MW (2006) Obscurin is required for the lateral alignment of striated myofibrils in zebrafish. Dev Dyn 235:2018–2029PubMedCrossRefGoogle Scholar
  45. Small JV, Fürst DO, Thornell L-E (1992) The cytoskeletal lattice of muscle cells. Eur J Biochem 208:559–572PubMedCrossRefGoogle Scholar
  46. Steiner F, Weber K, Fürst DO (1999) M band proteins myomesin and skelemin are encoded by the same gene: analysis of its organization and expression. Genomics 56:78–89PubMedCrossRefGoogle Scholar
  47. Street SF (1983) Lateral transmission of tension in frog myofibers: a myofibrillar network and transverse cytoskeletal connections are possible transmitters. J Cell Physiol 114:346–364PubMedCrossRefGoogle Scholar
  48. Thornell L-E, Eriksson A (1981) Filament system in the Purkinje fibers of the heart. Am J Physiol 241:H291–H305PubMedGoogle Scholar
  49. Thornell L-E, Eriksson A, Johansson B, Kjörell U, Franke WW, Virtanen I, Lehto VP (1985) Intermediate filament and associated proteins in heart Purkinje fibers: a membrane-myofibril anchored cytoskeletal system. Ann NY Acad Sci 455:213–240PubMedCrossRefGoogle Scholar
  50. Thornell LE, Yu JG, Carlsson L (2004) Z-disc streaming in muscle pathology —myofibrillar disruption or myofibrillar remodelling. Neuromuscul Disord 14:562–563Google Scholar
  51. Tidball JG (1992) Desmin at myotendinous junctions. Exp Cell Res 199:206–212PubMedCrossRefGoogle Scholar
  52. Wang K, Ramirez-Mitchell R (1983) A network of transverse and longitudinal intermediate filaments is associated with sarcomeres of adult vertebrate skeletal muscle. J Cell Biol 96:562–570PubMedCrossRefGoogle Scholar
  53. Young P, Ehler E, Gautel M, Sanger JW, Sanger JM (2001) Obscurin, a giant sarcomeric Rho guanine nucleotide exchange factor protein involved in sarcomere assembly. J Cell Biol 154:123–136PubMedCrossRefGoogle Scholar
  54. Yu JG, Malm C, Thornell LE (2002a) Eccentric contractions leading to DOMS do not cause loss of desmin nor fibre necrosis in human muscle. Histochem Cell Biol 118:29–34PubMedGoogle Scholar
  55. Yu JG, Thornell LE, Malm C (2002b) Desmin and actin alterations in human muscles affected by delayed onset muscle soreness: a high resolution immunocytochemical study. Histochem Cell Biol 118:171–179PubMedGoogle Scholar
  56. Yu JG, Fürst DO, Thornell LE (2003) The mode of myofibril remodelling in human skeletal muscle affected by DOMS induced by eccentric contractions. Histochem Cell Biol 119:383–393PubMedGoogle Scholar
  57. Yu JG, Carlsson L, Thornell LE (2004) Evidence for myofibril remodeling as opposed to myofibril damage in human muscles with DOMS: an ultrastructural and immunoelectron microscopic study. Histochem Cell Biol 121:219–227PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  1. 1.Department of Integrative Medical Biology, Section for AnatomyUmeå UniversityUmeåSweden
  2. 2.Department of Surgical and Perioperative Sciences, Sports MedicineUmeå UniversityUmeåSweden

Personalised recommendations