Advertisement

Histochemistry and Cell Biology

, Volume 131, Issue 3, pp 371–382 | Cite as

Localization of ank1.5 in the sarcoplasmic reticulum precedes that of SERCA and RyR: relationship with the organization of obscurin in developing sarcomeres

  • Emiliana Giacomello
  • Vincenzo SorrentinoEmail author
Original Paper

Abstract

Ank1.5 is a muscle-specific isoform of ankyrin1 localized on the sarcoplasmic reticulum (SR) membrane that has been shown to interact with obscurin, a sarcomeric protein. We report here studies on the localization of obscurin and ank1.5 in embryonic and postnatal rodent skeletal muscles. Using two antibodies against epitopes in the N- and C-terminus of obscurin, two distinct patterns of localization were observed. Before birth, the antibodies against the N- and the C-terminus of obscurin stained the Z-disk and M-band, respectively. At the same time, ank1.5 was detected at the Z-disk, rising the possibility that obscurin molecules at M-band may not be able to interact with ank1.5. Localization of ank1.5 at Z-disks in E14 muscle fibers revealed that ank1.5 is among the earliest SR proteins to assemble, since its organization preceded that of other SR proteins, like SERCA and RyR. After birth, the antibody against the N-terminus of obscurin stained the M-band while that against the C-terminus stained both M-bands and the Z-disks. Starting from postnatal day 1, ank1.5 was found at the level of both M-bands and Z-disks. Altogether, from these results we infer that exposure of some obscurin epitopes changes during skeletal muscle development, resulting in distinct, antibody-specific, localization pattern. Why this occurs is not clear, yet these data indicate that the organization of obscurin at different locations in the sarcomere changes during muscle development and that this might affect the interaction with ank1.5.

Keywords

Obscurin Small ankyrins Sarcoplasmic reticulum Skeletal muscle development 

Notes

Acknowledgments

We are grateful to Drs. Aikaterini Kontrogianni-Konstantopoulos and Robert J. Bloch (University of Maryland School of Medicine) for their generous gift of antibodies, useful discussions and for comments on the draft. We are also grateful to Dr Emilia Assenza for her contribution to the initial part of these experiments. This work was supported in part by grants from Agenzia Spaziale Italiana (ASI), the University of Siena (PAR 2006 and PAR 2007) and by Telethon (GGP08153).

References

  1. 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
  2. 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
  3. 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 Z-line to I-band linking system. Circ Res 89:1065–1072PubMedCrossRefGoogle Scholar
  4. Bennett V, Baines AJ (2001) Spectrin and ankyrin-based pathways: metazoan inventions for integrating cells into tissues. Physiol Rev 81:1353–1392PubMedGoogle Scholar
  5. 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
  6. 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 125:227–238PubMedCrossRefGoogle Scholar
  7. Borisov AB, Raeker MO, Russell MW (2008a) Developmental expression and differential cellular localization of obscurin and obscurin-associated kinase in cardiac muscle cells. J Cell Biochem 103:1621–1635PubMedCrossRefGoogle Scholar
  8. Borisov AB, Raeker MO, Russell MW (2008b) Early incorporation of obscurin into nascent sarcomeres: implication for myofibril assembly during cardiac myogenesis. Histochem Cell Biol 129:463–478PubMedCrossRefGoogle Scholar
  9. 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
  10. Carlsson L, Yu G, Thornell LE (2008) New aspects of obscurin in human striated muscles. Histochem Cell Biol 130:91–103PubMedCrossRefGoogle Scholar
  11. 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
  12. Dabiri GA, Turnacioglu KK, Sanger JM, Sanger JW (1997) Myofibrillogenesis visualized in living embryonic cardiomyocytes. Proc Natl Acad Sci USA 94:9493–9498PubMedCrossRefGoogle Scholar
  13. Flucher BE, Takekura H, Franzini-Armstrong C (1993) Development of the excitation–contraction coupling apparatus in skeletal muscle: association of sarcoplasmic reticulum and transverse tubules with myofibrils. Dev Biol 160:135–147PubMedCrossRefGoogle Scholar
  14. Franzini-Armstrong C (1994) The sarcoplasmic reticulum and the transverse tubules. In: Myology. Mc Graw-Hill, New York, pp 176–199Google Scholar
  15. Franzini-Armstrong C (2004) The membrane systems of muscle cells. In: Myology. Mc Graw-Hill, New York, pp 232–256Google Scholar
  16. Fukuzawa A, Idowu S, Gautel M (2006) Complete human gene structure of obscurin: implications for isoform generation by differential splicing. J Muscle Res Cell Motil 26:427–434CrossRefGoogle Scholar
  17. Fukuzawa A, Lange S, Holt M, Vihola A, Carmignac V, Ferreiro A, Udd B, Gautel M (2008) Interactions with titin and myomesin target obscurin and obscurin-like 1 to the M-band—implications for hereditary myopathies. J Cell Sci 121:1841–1851PubMedCrossRefGoogle Scholar
  18. Giannini G, Conti A, Mammarella S, Scrobogna M, Sorrentino V (1995) The ryanodine receptor/calcium channel genes are widely and differentially expressed in murine brain and peripheral tissues. J Cell Biol 128:893–904PubMedCrossRefGoogle Scholar
  19. Jorgensen AO, Kalnins VI, Zubrzycka E, MacLennan DH (1977) Assembly of the sarcoplasmic reticulum. Localization by immunofluorescence of sarcoplasmic reticulum proteins in differentiating rat skeletal muscle cell cultures. J Cell Biol 74:287–298PubMedCrossRefGoogle Scholar
  20. Kontrogianni-Konstantopoulos A, Bloch RJ (2005) Obscurin: a multitasking muscle giant. J Muscle Res Cell Motil 26:419–426PubMedCrossRefGoogle Scholar
  21. 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
  22. 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
  23. Kontrogianni-Konstantopoulos A, Catino DH, Strong JC, Sutter S, Borisov AB, Pumplin DW, Russell MW, Bloch RJ (2006) Obscurin modulates the assembly and organization of sarcomeres and the sarcoplasmic reticulum. FASEB J 20:2102–2111PubMedCrossRefGoogle Scholar
  24. Porter KR, Palade GE (1957) Studies on the endoplasmic reticulum. III. Its form and distribution in striated muscle cells. J Biophys Biochem Cytol 3:269–300PubMedCrossRefGoogle Scholar
  25. Porter NC, Resneck WG, O’Neill A, Van Rossum DB, Stone MR, Bloch RJ (2005) Association of small ankyrin 1 with the sarcoplasmic reticulum. Mol Membr Biol 22:421–432PubMedCrossRefGoogle Scholar
  26. Protasi F, Sun XH, Franzini-Armstrong C (1996) Formation and maturation of the calcium release apparatus in developing and adult avian myocardium. Dev Biol 173:265–278PubMedCrossRefGoogle Scholar
  27. Protasi F, Franzini-Armstrong C, Flucher BE (1997) Coordinated incorporation of skeletal muscle dihydropyridine receptors and ryanodine receptors in peripheral couplings of BC3H1 cells. J Cell Biol 137:859–870PubMedCrossRefGoogle Scholar
  28. Rossi D, Barone V, Giacomello E, Cusimano V, Sorrentino V (2008) The sarcoplasmic reticulum: an organized patchwork of specialized domains. Traffic 9:1044–1049PubMedCrossRefGoogle Scholar
  29. Russel MW, Raeker MO, Korytkowski KA, Sonneman KJ (2002) Identification, tissue expression and chromosomal localization of human Obscurin-MLCK, a member of the titin and dbl families of myosin light chain kinases. Gene 282:237–246CrossRefGoogle Scholar
  30. Sanger JM, Dome JS, Mittal B, Somlyo AV, Sanger JW (1989) Dynamics of the endoplasmic reticulum in living non-muscle and muscle cells. Cell Motil Cytoskeleton 13:301–319PubMedCrossRefGoogle Scholar
  31. Sanger JW, Chowrashi P, Shaner NC, Spalthoff S, Wang J, Freeman NL, Sanger JM (2002) Myofibrillogenesis in skeletal muscle cells. Clin Orthop Relat Res 403(Suppl):153–162CrossRefGoogle Scholar
  32. Schiaffino S, Margreth A (1969) Coordinated development of the sarcoplasmic reticulum and T system during postnatal differentiation of rat skeletal muscle. J Cell Biol 41:855–875PubMedCrossRefGoogle Scholar
  33. Schiaffino S, Reggiani C (1994) Myosin isoforms in mammalian skeletal muscle. J Appl Physiol 77:493–501PubMedGoogle Scholar
  34. Takekura H, Shuman H, Franzini-Armstrong C (1993) Differentiation of membrane systems during development of slow and fast skeletal muscle fibres in chicken. J Muscle Res Cell Motil 14:633–645PubMedCrossRefGoogle Scholar
  35. Takekura H, Flucher BE, Franzini-Armstrong C (2001) Sequential docking, molecular differentiation, and positioning of T-Tubule/SR junctions in developing mouse skeletal muscle. Dev Biol 239:204–214PubMedCrossRefGoogle Scholar
  36. Walker SM, Schrodt GR, Bingham M (1968) Electron microscope study of the sarcoplasmic reticulum at the Z line level in skeletal muscle fibers of fetal and newborn rats. J Cell Biol 39:469–475PubMedCrossRefGoogle Scholar
  37. Young P, Ehler E, Gautel M (2001) Obscurin, a giant sarcomeric Rho guanine nucleotide exchange factor protein involved in sarcomere assembly. J Cell Biol 154:123–136PubMedCrossRefGoogle Scholar
  38. Zhou D, Birkenmeier C-S, Williams M-W, Sharp J-J, Barker J-E, Bloch R-J (1997) Small, membrane-bound, alternatively spliced forms of ankyrin 1 associated with the sarcoplasmic reticulum of mammalian skeletal muscle. J Cell Biol 136:621–631PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  1. 1.Molecular Medicine Section, Department of Neuroscience and Interuniversity Institute of MyologyUniversity of SienaSienaItaly

Personalised recommendations