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. 1987 Dec 1;105(6):2457–2469. doi: 10.1083/jcb.105.6.2457

Agrin-like molecules at synaptic sites in normal, denervated, and damaged skeletal muscles

PMCID: PMC2114733  PMID: 2826488

Abstract

Several lines of evidence have led to the hypothesis that agrin, a protein extracted from the electric organ of Torpedo, is similar to the molecules in the synaptic cleft basal lamina at the neuromuscular junction that direct the formation of acetylcholine receptor and acetylcholinesterase aggregates on regenerating myofibers. One such finding is that monoclonal antibodies against agrin stain molecules concentrated in the synaptic cleft of neuromuscular junctions in rays. In the studies described here we made additional monoclonal antibodies against agrin and used them to extend our knowledge of agrin-like molecules at the neuromuscular junction. We found that anti-agrin antibodies intensely stained the synaptic cleft of frog and chicken as well as that of rays, that denervation of frog muscle resulted in a reduction in staining at the neuromuscular junction, and that the synaptic basal lamina in frog could be stained weeks after degeneration of all cellular components of the neuromuscular junction. We also describe anti-agrin staining in nonjunctional regions of muscle. We conclude the following: (a) agrin-like molecules are likely to be common to all vertebrate neuromuscular junctions; (b) the long-term maintenance of such molecules at the junction is nerve dependent; (c) the molecules are, indeed, a component of the synaptic basal lamina; and (d) they, like the molecules that direct the formation of receptor and esterase aggregates on regenerating myofibers, remain associated with the synaptic basal lamina after muscle damage.

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Selected References

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  1. Anglister L., McMahan U. J. Basal lamina directs acetylcholinesterase accumulation at synaptic sites in regenerating muscle. J Cell Biol. 1985 Sep;101(3):735–743. doi: 10.1083/jcb.101.3.735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brandan E., Maldonado M., Garrido J., Inestrosa N. C. Anchorage of collagen-tailed acetylcholinesterase to the extracellular matrix is mediated by heparan sulfate proteoglycans. J Cell Biol. 1985 Sep;101(3):985–992. doi: 10.1083/jcb.101.3.985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Burden S. J., Sargent P. B., McMahan U. J. Acetylcholine receptors in regenerating muscle accumulate at original synaptic sites in the absence of the nerve. J Cell Biol. 1979 Aug;82(2):412–425. doi: 10.1083/jcb.82.2.412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Carlson S. S., Caroni P., Kelly R. B. A nerve terminal anchorage protein from electric organ. J Cell Biol. 1986 Aug;103(2):509–520. doi: 10.1083/jcb.103.2.509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fallon J. R., Nitkin R. M., Reist N. E., Wallace B. G., McMahan U. J. Acetylcholine receptor-aggregating factor is similar to molecules concentrated at neuromuscular junctions. Nature. 1985 Jun 13;315(6020):571–574. doi: 10.1038/315571a0. [DOI] [PubMed] [Google Scholar]
  6. GINSBORG B. L. Spontaneous activity in muscle fibres of the chick. J Physiol. 1960 Mar;150:707–717. doi: 10.1113/jphysiol.1960.sp006413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Godfrey E. W., Nitkin R. M., Wallace B. G., Rubin L. L., McMahan U. J. Components of Torpedo electric organ and muscle that cause aggregation of acetylcholine receptors on cultured muscle cells. J Cell Biol. 1984 Aug;99(2):615–627. doi: 10.1083/jcb.99.2.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. LILEY A. W. An investigation of spontaneous activity at the neuromuscular junction of the rat. J Physiol. 1956 Jun 28;132(3):650–666. doi: 10.1113/jphysiol.1956.sp005555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Letinsky M. S., Fischbeck K. H., McMahan U. J. Precision of reinnervation of original postsynaptic sites in frog muscle after a nerve crush. J Neurocytol. 1976 Dec;5(6):691–718. doi: 10.1007/BF01181582. [DOI] [PubMed] [Google Scholar]
  10. MAURO A. Satellite cell of skeletal muscle fibers. J Biophys Biochem Cytol. 1961 Feb;9:493–495. doi: 10.1083/jcb.9.2.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Magill C., Reist N. E., Fallon J. R., Nitkin R. M., Wallace B. G., McMahan U. J. Agrin. Prog Brain Res. 1987;71:391–396. [PubMed] [Google Scholar]
  12. Marshall L. M., Sanes J. R., McMahan U. J. Reinnervation of original synaptic sites on muscle fiber basement membrane after disruption of the muscle cells. Proc Natl Acad Sci U S A. 1977 Jul;74(7):3073–3077. doi: 10.1073/pnas.74.7.3073. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. McLean I. W., Nakane P. K. Periodate-lysine-paraformaldehyde fixative. A new fixation for immunoelectron microscopy. J Histochem Cytochem. 1974 Dec;22(12):1077–1083. doi: 10.1177/22.12.1077. [DOI] [PubMed] [Google Scholar]
  14. McMahan U. J., Slater C. R. The influence of basal lamina on the accumulation of acetylcholine receptors at synaptic sites in regenerating muscle. J Cell Biol. 1984 Apr;98(4):1453–1473. doi: 10.1083/jcb.98.4.1453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nitkin R. M., Smith M. A., Magill C., Fallon J. R., Yao Y. M., Wallace B. G., McMahan U. J. Identification of agrin, a synaptic organizing protein from Torpedo electric organ. J Cell Biol. 1987 Dec;105(6 Pt 1):2471–2478. doi: 10.1083/jcb.105.6.2471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nitkin R. M., Wallace B. G., Spira M. E., Godfrey E. W., McMahan U. J. Molecular components of the synaptic basal lamina that direct differentiation of regenerating neuromuscular junctions. Cold Spring Harb Symp Quant Biol. 1983;48(Pt 2):653–665. doi: 10.1101/sqb.1983.048.01.069. [DOI] [PubMed] [Google Scholar]
  17. Page S. G. Structure and some contractile properties of fast and slow muscles of the chicken. J Physiol. 1969 Nov;205(1):131–145. doi: 10.1113/jphysiol.1969.sp008956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rieger F., Daniloff J. K., Pincon-Raymond M., Crossin K. L., Grumet M., Edelman G. M. Neuronal cell adhesion molecules and cytotactin are colocalized at the node of Ranvier. J Cell Biol. 1986 Aug;103(2):379–391. doi: 10.1083/jcb.103.2.379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Sealock R., Kavookjian A. Postsynaptic distribution of acetylcholine receptors in electroplax of the torpedine ray, Narcine brasiliensis. Brain Res. 1980 May 19;190(1):81–93. doi: 10.1016/0006-8993(80)91161-0. [DOI] [PubMed] [Google Scholar]
  21. Wallace B. G. Aggregating factor from Torpedo electric organ induces patches containing acetylcholine receptors, acetylcholinesterase, and butyrylcholinesterase on cultured myotubes. J Cell Biol. 1986 Mar;102(3):783–794. doi: 10.1083/jcb.102.3.783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wallace B. G., Nitkin R. M., Reist N. E., Fallon J. R., Moayeri N. N., McMahan U. J. Aggregates of acetylcholinesterase induced by acetylcholine receptor-aggregating factor. Nature. 1985 Jun 13;315(6020):574–577. doi: 10.1038/315574a0. [DOI] [PubMed] [Google Scholar]

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