Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1986 Mar 1;102(3):716–730. doi: 10.1083/jcb.102.3.716

Distribution of N-CAM in synaptic and extrasynaptic portions of developing and adult skeletal muscle

PMCID: PMC2114104  PMID: 3512581

Abstract

Previous studies of denervated and cultured muscle have shown that the expression of the neural cell adhesion molecule (N-CAM) in muscle is regulated by the muscle's state of innervation and that N-CAM might mediate some developmentally important nerve-muscle interactions. As a first step in learning whether N-CAM might regulate or be regulated by nerve-muscle interactions during normal development, we have used light and electron microscopic immunohistochemical methods to study its distribution in embryonic, perinatal, and adult rat muscle. In embryonic muscle, N-CAM is uniformly present on the surface of myotubes and in intramuscular nerves; N-CAM is also present on myoblasts, both in vivo and in cultures of embryonic muscle. N-CAM is lost from the nerves as myelination proceeds, and from myotubes as they mature. The loss of N-CAM from extrasynaptic portions of the myotube is a complex process, comprising a rapid rearrangement as secondary myotubes form, a phase of decline late in embryogenesis, a transient reappearance perinatally, and a more gradual disappearance during the first two postnatal weeks. Throughout embryonic and perinatal life, N-CAM is present at similar levels in synaptic and extrasynaptic regions of the myotube surface. However, N-CAM becomes concentrated in synaptic regions postnatally: it is present in postsynaptic and perisynaptic areas of the muscle fiber, both on the surface and intracellularly (in T-tubules), but undetectable in portions of muscle fibers distant from synapses. In addition, N-CAM is present on the surfaces of motor nerve terminals and of Schwann cells that cap nerve terminals, but absent from myelinated portions of motor axons and from myelinating Schwann cells. Thus, in the adult, N-CAM is present in synaptic but not extrasynaptic portions of all three cell types that comprise the neuromuscular junction. The times and places at which N-CAM appears are consistent with its playing several distinct roles in myogenesis, synaptogenesis, and synaptic maintenance, including alignment of secondary along primary myotubes, early interactions of axons with myotubes, and adhesion of Schwann cells to nerve terminals.

Full Text

The Full Text of this article is available as a PDF (2.2 MB).

Selected References

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

  1. Barnstable C. J., Dräger U. C. Thy-1 antigen: a ganglion cell specific marker in rodent retina. Neuroscience. 1984 Apr;11(4):847–855. doi: 10.1016/0306-4522(84)90195-7. [DOI] [PubMed] [Google Scholar]
  2. Bennett M. R. Development of neuromuscular synapses. Physiol Rev. 1983 Jul;63(3):915–1048. doi: 10.1152/physrev.1983.63.3.915. [DOI] [PubMed] [Google Scholar]
  3. Bevan S., Steinbach J. H. The distribution of alpha-bungarotoxin binding sites of mammalian skeletal muscle developing in vivo. J Physiol. 1977 May;267(1):195–213. doi: 10.1113/jphysiol.1977.sp011808. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burden S. Development of the neuromuscular junction in the chick embryo: the number, distribution, and stability of acetylcholine receptors. Dev Biol. 1977 Jun;57(2):317–329. doi: 10.1016/0012-1606(77)90218-4. [DOI] [PubMed] [Google Scholar]
  5. Carlson B. M., Faulkner J. A. The regeneration of skeletal muscle fibers following injury: a review. Med Sci Sports Exerc. 1983;15(3):187–198. [PubMed] [Google Scholar]
  6. Chiu A. Y., Sanes J. R. Development of basal lamina in synaptic and extrasynaptic portions of embryonic rat muscle. Dev Biol. 1984 Jun;103(2):456–467. doi: 10.1016/0012-1606(84)90333-6. [DOI] [PubMed] [Google Scholar]
  7. Covault J., Merlie J. P., Goridis C., Sanes J. R. Molecular forms of N-CAM and its RNA in developing and denervated skeletal muscle. J Cell Biol. 1986 Mar;102(3):731–739. doi: 10.1083/jcb.102.3.731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Covault J., Sanes J. R. Neural cell adhesion molecule (N-CAM) accumulates in denervated and paralyzed skeletal muscles. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4544–4548. doi: 10.1073/pnas.82.13.4544. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dennis M. J., Ziskind-Conhaim L., Harris A. J. Development of neuromuscular junctions in rat embryos. Dev Biol. 1981 Jan 30;81(2):266–279. doi: 10.1016/0012-1606(81)90290-6. [DOI] [PubMed] [Google Scholar]
  10. Edelman G. M. Cell adhesion molecules. Science. 1983 Feb 4;219(4584):450–457. doi: 10.1126/science.6823544. [DOI] [PubMed] [Google Scholar]
  11. Fambrough D. M. Control of acetylcholine receptors in skeletal muscle. Physiol Rev. 1979 Jan;59(1):165–227. doi: 10.1152/physrev.1979.59.1.165. [DOI] [PubMed] [Google Scholar]
  12. Fertuck H. C., Salpeter M. M. Quantitation of junctional and extrajunctional acetylcholine receptors by electron microscope autoradiography after 125I-alpha-bungarotoxin binding at mouse neuromuscular junctions. J Cell Biol. 1976 Apr;69(1):144–158. doi: 10.1083/jcb.69.1.144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gambke B., Rubinstein N. A. A monoclonal antibody to the embryonic myosin heavy chain of rat skeletal muscle. J Biol Chem. 1984 Oct 10;259(19):12092–12100. [PubMed] [Google Scholar]
  14. Gennarini G., Rougon G., Deagostini-Bazin H., Hirn M., Goridis C. Studies on the transmembrane disposition of the neural cell adhesion molecule N-CAM. A monoclonal antibody recognizing a cytoplasmic domain and evidence for the presence of phosphoserine residues. Eur J Biochem. 1984 Jul 2;142(1):57–64. doi: 10.1111/j.1432-1033.1984.tb08250.x. [DOI] [PubMed] [Google Scholar]
  15. Glicksman M. A., Willard M. Differential expression of the three neurofilament polypeptides. Ann N Y Acad Sci. 1985;455:479–491. doi: 10.1111/j.1749-6632.1985.tb50430.x. [DOI] [PubMed] [Google Scholar]
  16. Grumet M., Rutishauser U., Edelman G. M. Neural cell adhesion molecule is on embryonic muscle cells and mediates adhesion to nerve cells in vitro. Nature. 1982 Feb 25;295(5851):693–695. doi: 10.1038/295693a0. [DOI] [PubMed] [Google Scholar]
  17. Johnson G. D., Nogueira Araujo G. M. A simple method of reducing the fading of immunofluorescence during microscopy. J Immunol Methods. 1981;43(3):349–350. doi: 10.1016/0022-1759(81)90183-6. [DOI] [PubMed] [Google Scholar]
  18. Kelly A. M., Zacks S. I. The fine structure of motor endplate morphogenesis. J Cell Biol. 1969 Jul;42(1):154–169. doi: 10.1083/jcb.42.1.154. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kelly A. M., Zacks S. I. The histogenesis of rat intercostal muscle. J Cell Biol. 1969 Jul;42(1):135–153. doi: 10.1083/jcb.42.1.135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kohn J., Wilchek M. A new approach (cyano-transfer) for cyanogen bromide activation of Sepharose at neutral pH, which yields activated resins, free of interfering nitrogen derivatives. Biochem Biophys Res Commun. 1982 Aug;107(3):878–884. doi: 10.1016/0006-291x(82)90604-0. [DOI] [PubMed] [Google Scholar]
  21. Lemmon V., Staros E. B., Perry H. E., Gottlieb D. I. A monoclonal antibody which binds to the surface of chick brain cells and myotubes: cell selectivity and properties of the antigen. Brain Res. 1982 Mar;255(3):349–360. doi: 10.1016/0165-3806(82)90003-7. [DOI] [PubMed] [Google Scholar]
  22. Merlie J. P., Sanes J. R. Concentration of acetylcholine receptor mRNA in synaptic regions of adult muscle fibres. Nature. 1985 Sep 5;317(6032):66–68. doi: 10.1038/317066a0. [DOI] [PubMed] [Google Scholar]
  23. Miledi R., Slater C. R. Electrophysiology and electron-microscopy of rat neuromuscular junctions after nerve degeneration. Proc R Soc Lond B Biol Sci. 1968 Feb 27;169(1016):289–306. doi: 10.1098/rspb.1968.0012. [DOI] [PubMed] [Google Scholar]
  24. Moore S. E., Walsh F. S. Specific regulation of N-CAM/D2-CAM cell adhesion molecule during skeletal muscle development. EMBO J. 1985 Mar;4(3):623–630. doi: 10.1002/j.1460-2075.1985.tb03675.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Morris R. J., Barber P. C., Beech J., Raisman G. The distribution of Thy-1 antigen in the P.N.S. of the adult rat. J Neurocytol. 1983 Dec;12(6):1017–1039. doi: 10.1007/BF01153348. [DOI] [PubMed] [Google Scholar]
  26. Ontell M., Kozeka K. The organogenesis of murine striated muscle: a cytoarchitectural study. Am J Anat. 1984 Oct;171(2):133–148. doi: 10.1002/aja.1001710202. [DOI] [PubMed] [Google Scholar]
  27. Pestronk A. Intracellular acetylcholine receptors in skeletal muscles of the adult rat. J Neurosci. 1985 May;5(5):1111–1117. doi: 10.1523/JNEUROSCI.05-05-01111.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Ravdin P., Axelrod D. Fluorescent tetramethyl rhodamine derivatives of alpha-bungarotoxin: preparation, separation, and characterization. Anal Biochem. 1977 Jun;80(2):585–592. doi: 10.1016/0003-2697(77)90682-0. [DOI] [PubMed] [Google Scholar]
  29. Rieger F., Grumet M., Edelman G. M. N-CAM at the vertebrate neuromuscular junction. J Cell Biol. 1985 Jul;101(1):285–293. doi: 10.1083/jcb.101.1.285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rutishauser U. Developmental biology of a neural cell adhesion molecule. Nature. 1984 Aug 16;310(5978):549–554. doi: 10.1038/310549a0. [DOI] [PubMed] [Google Scholar]
  31. Rutishauser U., Grumet M., Edelman G. M. Neural cell adhesion molecule mediates initial interactions between spinal cord neurons and muscle cells in culture. J Cell Biol. 1983 Jul;97(1):145–152. doi: 10.1083/jcb.97.1.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rutishauser U., Hoffman S., Edelman G. M. Binding properties of a cell adhesion molecule from neural tissue. Proc Natl Acad Sci U S A. 1982 Jan;79(2):685–689. doi: 10.1073/pnas.79.2.685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sanes J. R., Cheney J. M. Lectin binding reveals a synapse-specific carbohydrate in skeletal muscle. Nature. 1982 Dec 16;300(5893):646–647. doi: 10.1038/300646a0. [DOI] [PubMed] [Google Scholar]
  34. Sanes J. R., Chiu A. Y. The basal lamina of the neuromuscular junction. Cold Spring Harb Symp Quant Biol. 1983;48(Pt 2):667–678. doi: 10.1101/sqb.1983.048.01.070. [DOI] [PubMed] [Google Scholar]
  35. Sanes J. R., Feldman D. H., Cheney J. M., Lawrence J. C., Jr Brain extract induces synaptic characteristics in the basal lamina of cultured myotubes. J Neurosci. 1984 Feb;4(2):464–473. doi: 10.1523/JNEUROSCI.04-02-00464.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sanes J. R., Hall Z. W. Antibodies that bind specifically to synaptic sites on muscle fiber basal lamina. J Cell Biol. 1979 Nov;83(2 Pt 1):357–370. doi: 10.1083/jcb.83.2.357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sanes J. R., Lawrence J. C., Jr Activity-dependent accumulation of basal lamina by cultured rat myotubes. Dev Biol. 1983 May;97(1):123–136. doi: 10.1016/0012-1606(83)90070-2. [DOI] [PubMed] [Google Scholar]
  38. Sanes J. R., Marshall L. M., McMahan U. J. Reinnervation of muscle fiber basal lamina after removal of myofibers. Differentiation of regenerating axons at original synaptic sites. J Cell Biol. 1978 Jul;78(1):176–198. doi: 10.1083/jcb.78.1.176. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Sanes J. R., Schachner M., Covault J. Expression of several adhesive macromolecules (N-CAM, L1, J1, NILE, uvomorulin, laminin, fibronectin, and a heparan sulfate proteoglycan) in embryonic, adult, and denervated adult skeletal muscle. J Cell Biol. 1986 Feb;102(2):420–431. doi: 10.1083/jcb.102.2.420. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sargent P. B., Hedges B. E., Tsavaler L., Clemmons L., Tzartos S., Lindstrom J. M. Structure and transmembrane nature of the acetylcholine receptor in amphibian skeletal muscle as revealed by cross-reacting monoclonal antibodies. J Cell Biol. 1984 Feb;98(2):609–618. doi: 10.1083/jcb.98.2.609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Schneider C., Newman R. A., Sutherland D. R., Asser U., Greaves M. F. A one-step purification of membrane proteins using a high efficiency immunomatrix. J Biol Chem. 1982 Sep 25;257(18):10766–10769. [PubMed] [Google Scholar]
  42. Wakelam M. J. The fusion of myoblasts. Biochem J. 1985 May 15;228(1):1–12. doi: 10.1042/bj2280001. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES