Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1991 Oct 2;115(2):411–421. doi: 10.1083/jcb.115.2.411

The subcellular distribution of dystrophin in mouse skeletal, cardiac, and smooth muscle

PMCID: PMC2289158  PMID: 1918148

Abstract

We use a highly specific and sensitive antibody to further characterize the distribution of dystrophin in skeletal, cardiac, and smooth muscle. No evidence for localization other than at the cell surface is apparent in skeletal muscle and no 427-kD dystrophin labeling was detected in sciatic nerve. An elevated concentration of dystrophin appears at the myotendinous junction and the neuromuscular junction, labeling in the latter being more intense specifically in the troughs of the synaptic folds. In cardiac muscle the distribution of dystrophin is limited to the surface plasma membrane but is notably absent from the membrane that overlays adherens junctions of the intercalated disks. In smooth muscle, the plasma membrane labeling is considerably less abundant than in cardiac or skeletal muscle and is found in areas of membrane underlain by membranous vesicles. As in cardiac muscle, smooth muscle dystrophin seems to be excluded from membrane above densities that mark adherens junctions. Dystrophin appears as a doublet on Western blots of skeletal and cardiac muscle, and as a single band of lower abundance in smooth muscle that corresponds most closely in molecular weight to the upper band of the striated muscle doublet. The lower band of the doublet in striated muscle appears to lack a portion of the carboxyl terminus and may represent a dystrophin isoform. Isoform differences and the presence of dystrophin on different specialized membrane surfaces imply multiple functional roles for the dystrophin protein.

Full Text

The Full Text of this article is available as a PDF (1.6 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arahata K., Ishiura S., Ishiguro T., Tsukahara T., Suhara Y., Eguchi C., Ishihara T., Nonaka I., Ozawa E., Sugita H. Immunostaining of skeletal and cardiac muscle surface membrane with antibody against Duchenne muscular dystrophy peptide. Nature. 1988 Jun 30;333(6176):861–863. doi: 10.1038/333861a0. [DOI] [PubMed] [Google Scholar]
  2. Bar S., Barnea E., Levy Z., Neuman S., Yaffe D., Nudel U. A novel product of the Duchenne muscular dystrophy gene which greatly differs from the known isoforms in its structure and tissue distribution. Biochem J. 1990 Dec 1;272(2):557–560. doi: 10.1042/bj2720557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bennett V. Spectrin-based membrane skeleton: a multipotential adaptor between plasma membrane and cytoplasm. Physiol Rev. 1990 Oct;70(4):1029–1065. doi: 10.1152/physrev.1990.70.4.1029. [DOI] [PubMed] [Google Scholar]
  4. Bonilla E., Samitt C. E., Miranda A. F., Hays A. P., Salviati G., DiMauro S., Kunkel L. M., Hoffman E. P., Rowland L. P. Duchenne muscular dystrophy: deficiency of dystrophin at the muscle cell surface. Cell. 1988 Aug 12;54(4):447–452. doi: 10.1016/0092-8674(88)90065-7. [DOI] [PubMed] [Google Scholar]
  5. Brown R. H., Jr, Hoffman E. P. Molecular biology of Duchenne muscular dystrophy. Trends Neurosci. 1988 Nov;11(11):480–484. doi: 10.1016/0166-2236(88)90006-9. [DOI] [PubMed] [Google Scholar]
  6. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  7. Byers T. J., Husain-Chishti A., Dubreuil R. R., Branton D., Goldstein L. S. Sequence similarity of the amino-terminal domain of Drosophila beta spectrin to alpha actinin and dystrophin. J Cell Biol. 1989 Oct;109(4 Pt 1):1633–1641. doi: 10.1083/jcb.109.4.1633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Campbell K. P., Kahl S. D. Association of dystrophin and an integral membrane glycoprotein. Nature. 1989 Mar 16;338(6212):259–262. doi: 10.1038/338259a0. [DOI] [PubMed] [Google Scholar]
  9. Carpenter S., Karpati G., Zubrzycka-Gaarn E., Bulman D. E., Ray P. N., Worton R. G. Dystrophin is localized to the plasma membrane of human skeletal muscle fibers by electron-microscopic cytochemical study. Muscle Nerve. 1990 May;13(5):376–380. doi: 10.1002/mus.880130503. [DOI] [PubMed] [Google Scholar]
  10. Chelly J., Kaplan J. C., Maire P., Gautron S., Kahn A. Transcription of the dystrophin gene in human muscle and non-muscle tissue. Nature. 1988 Jun 30;333(6176):858–860. doi: 10.1038/333858a0. [DOI] [PubMed] [Google Scholar]
  11. Cullen M. J., Walsh J., Nicholson L. V., Harris J. B. Ultrastructural localization of dystrophin in human muscle by using gold immunolabelling. Proc R Soc Lond B Biol Sci. 1990 May 22;240(1297):197–210. doi: 10.1098/rspb.1990.0034. [DOI] [PubMed] [Google Scholar]
  12. Devine C. E., Simpson F. O., Bertaud W. S. Surface features of smooth muscle cells from the mesenteric artery and vas deferens. J Cell Sci. 1971 Mar;8(2):427–443. doi: 10.1242/jcs.8.2.427. [DOI] [PubMed] [Google Scholar]
  13. Drenckhahn D., Beckerle M., Burridge K., Otto J. Identification and subcellular location of talin in various cell types and tissues by means of [125I]vinculin overlay, immunoblotting and immunocytochemistry. Eur J Cell Biol. 1988 Aug;46(3):513–522. [PubMed] [Google Scholar]
  14. Fardeau M., Tomé F. M., Collin H., Augier N., Pons F., Léger J., Léger J. Présence d'une protéine de type dystrophine au niveau de la jonction neuromusculaire dans la dystrophie musculaire de Duchenne et la souris mutante "mdx". C R Acad Sci III. 1990;311(5):197–204. [PubMed] [Google Scholar]
  15. Feener C. A., Koenig M., Kunkel L. M. Alternative splicing of human dystrophin mRNA generates isoforms at the carboxy terminus. Nature. 1989 Apr 6;338(6215):509–511. doi: 10.1038/338509a0. [DOI] [PubMed] [Google Scholar]
  16. Flucher B. E., Daniels M. P. Distribution of Na+ channels and ankyrin in neuromuscular junctions is complementary to that of acetylcholine receptors and the 43 kd protein. Neuron. 1989 Aug;3(2):163–175. doi: 10.1016/0896-6273(89)90029-9. [DOI] [PubMed] [Google Scholar]
  17. Geiger B., Tokuyasu K. T., Dutton A. H., Singer S. J. Vinculin, an intracellular protein localized at specialized sites where microfilament bundles terminate at cell membranes. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4127–4131. doi: 10.1073/pnas.77.7.4127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hammonds R. G., Jr Protein sequence of DMD gene is related to actin-binding domain of alpha-actinin. Cell. 1987 Oct 9;51(1):1–1. doi: 10.1016/0092-8674(87)90002-x. [DOI] [PubMed] [Google Scholar]
  19. Harris A. S., Morrow J. S. Calmodulin and calcium-dependent protease I coordinately regulate the interaction of fodrin with actin. Proc Natl Acad Sci U S A. 1990 Apr;87(8):3009–3013. doi: 10.1073/pnas.87.8.3009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hoffman E. P., Brown R. H., Jr, Kunkel L. M. Dystrophin: the protein product of the Duchenne muscular dystrophy locus. Cell. 1987 Dec 24;51(6):919–928. doi: 10.1016/0092-8674(87)90579-4. [DOI] [PubMed] [Google Scholar]
  21. Hoffman E. P., Fischbeck K. H., Brown R. H., Johnson M., Medori R., Loike J. D., Harris J. B., Waterston R., Brooke M., Specht L. Characterization of dystrophin in muscle-biopsy specimens from patients with Duchenne's or Becker's muscular dystrophy. N Engl J Med. 1988 May 26;318(21):1363–1368. doi: 10.1056/NEJM198805263182104. [DOI] [PubMed] [Google Scholar]
  22. Hoffman E. P., Hudecki M. S., Rosenberg P. A., Pollina C. M., Kunkel L. M. Cell and fiber-type distribution of dystrophin. Neuron. 1988 Jul;1(5):411–420. doi: 10.1016/0896-6273(88)90191-2. [DOI] [PubMed] [Google Scholar]
  23. Hoffman E. P., Morgan J. E., Watkins S. C., Partridge T. A. Somatic reversion/suppression of the mouse mdx phenotype in vivo. J Neurol Sci. 1990 Oct;99(1):9–25. doi: 10.1016/0022-510x(90)90195-s. [DOI] [PubMed] [Google Scholar]
  24. Huard J., Fortier L. P., Labrecque C., Dansereau G., Tremblay J. P. Is dystrophin present in the nerve terminal at the neuromuscular junction? An immunohistochemical study of the heterozygote dystrophic (mdx) mouse. Synapse. 1991 Feb;7(2):135–140. doi: 10.1002/syn.890070207. [DOI] [PubMed] [Google Scholar]
  25. Jerusalem F., Engel A. G., Gomez M. R. Duchenne dystrophy. II. Morphometric study of motor end-plate fine structure. Brain. 1974 Mar;97(1):123–130. doi: 10.1093/brain/97.1.123. [DOI] [PubMed] [Google Scholar]
  26. Khurana T. S., Hoffman E. P., Kunkel L. M. Identification of a chromosome 6-encoded dystrophin-related protein. J Biol Chem. 1990 Oct 5;265(28):16717–16720. [PubMed] [Google Scholar]
  27. Knudson C. M., Hoffman E. P., Kahl S. D., Kunkel L. M., Campbell K. P. Evidence for the association of dystrophin with the transverse tubular system in skeletal muscle. J Biol Chem. 1988 Jun 15;263(17):8480–8484. [PubMed] [Google Scholar]
  28. Koenig M., Kunkel L. M. Detailed analysis of the repeat domain of dystrophin reveals four potential hinge segments that may confer flexibility. J Biol Chem. 1990 Mar 15;265(8):4560–4566. [PubMed] [Google Scholar]
  29. Koenig M., Monaco A. P., Kunkel L. M. The complete sequence of dystrophin predicts a rod-shaped cytoskeletal protein. Cell. 1988 Apr 22;53(2):219–228. doi: 10.1016/0092-8674(88)90383-2. [DOI] [PubMed] [Google Scholar]
  30. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  31. Lidov H. G., Byers T. J., Watkins S. C., Kunkel L. M. Localization of dystrophin to postsynaptic regions of central nervous system cortical neurons. Nature. 1990 Dec 20;348(6303):725–728. doi: 10.1038/348725a0. [DOI] [PubMed] [Google Scholar]
  32. Love D. R., Hill D. F., Dickson G., Spurr N. K., Byth B. C., Marsden R. F., Walsh F. S., Edwards Y. H., Davies K. E. An autosomal transcript in skeletal muscle with homology to dystrophin. Nature. 1989 May 4;339(6219):55–58. doi: 10.1038/339055a0. [DOI] [PubMed] [Google Scholar]
  33. Miike T., Miyatake M., Zhao J., Yoshioka K., Uchino M. Immunohistochemical dystrophin reaction in synaptic regions. Brain Dev. 1989;11(5):344–346. doi: 10.1016/s0387-7604(89)80067-1. [DOI] [PubMed] [Google Scholar]
  34. Nakamura K., Tanaka T., Kuwahara A., Takeo K. Microassay for proteins on nitrocellulose filter using protein dye-staining procedure. Anal Biochem. 1985 Aug 1;148(2):311–319. doi: 10.1016/0003-2697(85)90234-9. [DOI] [PubMed] [Google Scholar]
  35. Nudel U., Zuk D., Einat P., Zeelon E., Levy Z., Neuman S., Yaffe D. Duchenne muscular dystrophy gene product is not identical in muscle and brain. Nature. 1989 Jan 5;337(6202):76–78. doi: 10.1038/337076a0. [DOI] [PubMed] [Google Scholar]
  36. Ohlendieck K., Ervasti J. M., Snook J. B., Campbell K. P. Dystrophin-glycoprotein complex is highly enriched in isolated skeletal muscle sarcolemma. J Cell Biol. 1991 Jan;112(1):135–148. doi: 10.1083/jcb.112.1.135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Rowland L. P. Biochemistry of muscle membranes in Duchenne muscular dystrophy. Muscle Nerve. 1980 Jan-Feb;3(1):3–20. doi: 10.1002/mus.880030103. [DOI] [PubMed] [Google Scholar]
  38. Sakakibara H., Engel A. G., Lambert E. H. Duchenne dystrophy: ultrastructural localization of the acetylcholine receptor and intracellular microelectrode studies of neuromuscular transmission. Neurology. 1977 Aug;27(8):741–745. doi: 10.1212/wnl.27.8.741. [DOI] [PubMed] [Google Scholar]
  39. Salviati G., Betto R., Ceoldo S., Biasia E., Bonilla E., Miranda A. F., Dimauro S. Cell fractionation studies indicate that dystrophin is a protein of surface membranes of skeletal muscle. Biochem J. 1989 Mar 15;258(3):837–841. doi: 10.1042/bj2580837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Samitt C. E., Bonilla E. Immunocytochemical study of dystrophin at the myotendinous junction. Muscle Nerve. 1990 Jun;13(6):493–500. doi: 10.1002/mus.880130605. [DOI] [PubMed] [Google Scholar]
  41. Sealock R., Butler M. H., Kramarcy N. R., Gao K. X., Murnane A. A., Douville K., Froehner S. C. Localization of dystrophin relative to acetylcholine receptor domains in electric tissue and adult and cultured skeletal muscle. J Cell Biol. 1991 Jun;113(5):1133–1144. doi: 10.1083/jcb.113.5.1133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sicinski P., Geng Y., Ryder-Cook A. S., Barnard E. A., Darlison M. G., Barnard P. J. The molecular basis of muscular dystrophy in the mdx mouse: a point mutation. Science. 1989 Jun 30;244(4912):1578–1580. doi: 10.1126/science.2662404. [DOI] [PubMed] [Google Scholar]
  43. Small J. V. Geometry of actin-membrane attachments in the smooth muscle cell: the localisations of vinculin and alpha-actinin. EMBO J. 1985 Jan;4(1):45–49. doi: 10.1002/j.1460-2075.1985.tb02315.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Srinivasan Y., Elmer L., Davis J., Bennett V., Angelides K. Ankyrin and spectrin associate with voltage-dependent sodium channels in brain. Nature. 1988 May 12;333(6169):177–180. doi: 10.1038/333177a0. [DOI] [PubMed] [Google Scholar]
  45. Tidball J. G., Law D. J. Dystrophin is required for normal thin filament-membrane associations at myotendinous junctions. Am J Pathol. 1991 Jan;138(1):17–21. [PMC free article] [PubMed] [Google Scholar]
  46. Torres L. F., Duchen L. W. The mutant mdx: inherited myopathy in the mouse. Morphological studies of nerves, muscles and end-plates. Brain. 1987 Apr;110(Pt 2):269–299. doi: 10.1093/brain/110.2.269. [DOI] [PubMed] [Google Scholar]
  47. Watkins S. C., Hoffman E. P., Slayter H. S., Kunkel L. M. Immunoelectron microscopic localization of dystrophin in myofibres. Nature. 1988 Jun 30;333(6176):863–866. doi: 10.1038/333863a0. [DOI] [PubMed] [Google Scholar]
  48. Zubrzycka-Gaarn E. E., Bulman D. E., Karpati G., Burghes A. H., Belfall B., Klamut H. J., Talbot J., Hodges R. S., Ray P. N., Worton R. G. The Duchenne muscular dystrophy gene product is localized in sarcolemma of human skeletal muscle. Nature. 1988 Jun 2;333(6172):466–469. doi: 10.1038/333466a0. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES