Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1988 Nov 1;107(5):1825–1833. doi: 10.1083/jcb.107.5.1825

Motor neurons contain agrin-like molecules

PMCID: PMC2115317  PMID: 2846587

Abstract

Molecules antigenically similar to agrin, a protein extracted from the electric organ of Torpedo californica, are highly concentrated in the synaptic basal lamina of neuromuscular junctions in vertebrate skeletal muscle. On the basis of several lines of evidence it has been proposed that agrin-like molecules mediate the nerve-induced formation of acetylcholine receptor (AChR) and acetylcholinesterase (AChE) aggregates on the surface of muscle fibers at developing and regenerating neuromuscular junctions and that they help maintain these postsynaptic specializations in the adult. Here we show that anti-agrin monoclonal antibodies selectively stain the cell bodies of motor neurons in embryos and adults, and that the stain is concentrated in the Golgi apparatus. We also present evidence that motor neurons in both embryos and adults contain molecules that cause the formation of AChR and AChE aggregates on cultured myotubes and that these AChR/AChE- aggregating molecules are antigenically similar to agrin. These findings are consistent with the hypothesis that agrin-like molecules are synthesized by motor neurons, and are released from their axon terminals to become incorporated into the synaptic basal lamina where they direct the formation of synapses during development and regeneration.

Full Text

The Full Text of this article is available as a PDF (2.8 MB).

Selected References

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

  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. Bandtlow C. E., Heumann R., Schwab M. E., Thoenen H. Cellular localization of nerve growth factor synthesis by in situ hybridization. EMBO J. 1987 Apr;6(4):891–899. doi: 10.1002/j.1460-2075.1987.tb04835.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bauer H. C., Daniels M. P., Pudimat P. A., Jacques L., Sugiyama H., Christian C. N. Characterization and partial purification of a neuronal factor which increases acetylcholine receptor aggregation on cultured muscle cells. Brain Res. 1981 Mar 30;209(2):395–404. doi: 10.1016/0006-8993(81)90161-x. [DOI] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  5. Brown W. J., Farquhar M. G. The mannose-6-phosphate receptor for lysosomal enzymes is concentrated in cis Golgi cisternae. Cell. 1984 Feb;36(2):295–307. doi: 10.1016/0092-8674(84)90223-x. [DOI] [PubMed] [Google Scholar]
  6. Claude P., Hawrot E., Dunis D. A., Campenot R. B. Binding, internalization, and retrograde transport of 125I-nerve growth factor in cultured rat sympathetic neurons. J Neurosci. 1982 Apr;2(4):431–442. doi: 10.1523/JNEUROSCI.02-04-00431.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dennis M. J. Development of the neuromuscular junction: inductive interactions between cells. Annu Rev Neurosci. 1981;4:43–68. doi: 10.1146/annurev.ne.04.030181.000355. [DOI] [PubMed] [Google Scholar]
  8. Dohrmann U., Edgar D., Sendtner M., Thoenen H. Muscle-derived factors that support survival and promote fiber outgrowth from embryonic chick spinal motor neurons in culture. Dev Biol. 1986 Nov;118(1):209–221. doi: 10.1016/0012-1606(86)90089-8. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Fontaine B., Klarsfeld A., Hökfelt T., Changeux J. P. Calcitonin gene-related peptide, a peptide present in spinal cord motoneurons, increases the number of acetylcholine receptors in primary cultures of chick embryo myotubes. Neurosci Lett. 1986 Oct 30;71(1):59–65. doi: 10.1016/0304-3940(86)90257-0. [DOI] [PubMed] [Google Scholar]
  11. Godfrey E. W., Dietz M. E., Morstad A. L., Wallskog P. A., Yorde D. E. Acetylcholine receptor-aggregating proteins are associated with the extracellular matrix of many tissues in Torpedo. J Cell Biol. 1988 Apr;106(4):1263–1272. doi: 10.1083/jcb.106.4.1263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Gonatas N. K., Steiber A., Kim S. U., Graham D. I., Avrameas S. Internalization of neuronal plasma membrane ricin receptors into the Golgi apparatus. Exp Cell Res. 1975 Sep;94(2):426–431. doi: 10.1016/0014-4827(75)90508-x. [DOI] [PubMed] [Google Scholar]
  14. Harris A. J., Kuffler S. W., Dennis M. J. Differential chemosensitivity of synaptic and extrasynaptic areas on the neuronal surface membrane in parasympathetic neurons of the frog, tested by microapplication of acetylcholine. Proc R Soc Lond B Biol Sci. 1971 Apr 27;177(1049):541–553. doi: 10.1098/rspb.1971.0046. [DOI] [PubMed] [Google Scholar]
  15. Jacob M. H., Berg D. K., Lindstrom J. M. Shared antigenic determinant between the Electrophorus acetylcholine receptor and a synaptic component on chicken ciliary ganglion neurons. Proc Natl Acad Sci U S A. 1984 May;81(10):3223–3227. doi: 10.1073/pnas.81.10.3223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jacob M. H., Lindstrom J. M., Berg D. K. Surface and intracellular distribution of a putative neuronal nicotinic acetylcholine receptor. J Cell Biol. 1986 Jul;103(1):205–214. doi: 10.1083/jcb.103.1.205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Landmesser L., Morris D. G. The development of functional innervation in the hind limb of the chick embryo. J Physiol. 1975 Jul;249(2):301–326. doi: 10.1113/jphysiol.1975.sp011017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Loring R. H., Zigmond R. E. Ultrastructural distribution of 125I-toxin F binding sites on chick ciliary neurons: synaptic localization of a toxin that blocks ganglionic nicotinic receptors. J Neurosci. 1987 Jul;7(7):2153–2162. doi: 10.1523/JNEUROSCI.07-07-02153.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Marshall L. M. Synaptic localization of alpha-bungarotoxin binding which blocks nicotinic transmission at frog sympathetic neurons. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1948–1952. doi: 10.1073/pnas.78.3.1948. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. New H. V., Mudge A. W. Calcitonin gene-related peptide regulates muscle acetylcholine receptor synthesis. 1986 Oct 30-Nov 5Nature. 323(6091):809–811. doi: 10.1038/323809a0. [DOI] [PubMed] [Google Scholar]
  22. Newman E. A. High potassium conductance in astrocyte endfeet. Science. 1986 Jul 25;233(4762):453–454. doi: 10.1126/science.3726539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Podleski T. R., Axelrod D., Ravdin P., Greenberg I., Johnson M. M., Salpeter M. M. Nerve extract induces increase and redistribution of acetylcholine receptors on cloned muscle cells. Proc Natl Acad Sci U S A. 1978 Apr;75(4):2035–2039. doi: 10.1073/pnas.75.4.2035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Reist N. E., Magill C., McMahan U. J. Agrin-like molecules at synaptic sites in normal, denervated, and damaged skeletal muscles. J Cell Biol. 1987 Dec;105(6 Pt 1):2457–2469. doi: 10.1083/jcb.105.6.2457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rotundo R. L. Purification and properties of the membrane-bound form of acetylcholinesterase from chicken brain. Evidence for two distinct polypeptide chains. J Biol Chem. 1984 Nov 10;259(21):13186–13194. [PubMed] [Google Scholar]
  27. 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]
  28. Schaffner A. E., Daniels M. P. Conditioned medium from cultures of embryonic neurons contains a high molecular weight factor which induces acetylcholine receptor aggregation on cultured myotubes. J Neurosci. 1982 May;2(5):623–632. doi: 10.1523/JNEUROSCI.02-05-00623.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Schwab M. E. Ultrastructural localization of a nerve growth factor-horseradish peroxidase (NGF-HRP) coupling product after retrograde axonal transport in adrenergic neurons. Brain Res. 1977 Jul 8;130(1):190–196. doi: 10.1016/0006-8993(77)90857-5. [DOI] [PubMed] [Google Scholar]
  30. Smith M. A., Yao Y. M., Reist N. E., Magill C., Wallace B. G., McMahan U. J. Identification of agrin in electric organ extracts and localization of agrin-like molecules in muscle and central nervous system. J Exp Biol. 1987 Sep;132:223–230. doi: 10.1242/jeb.132.1.223. [DOI] [PubMed] [Google Scholar]
  31. Usdin T. B., Fischbach G. D. Purification and characterization of a polypeptide from chick brain that promotes the accumulation of acetylcholine receptors in chick myotubes. J Cell Biol. 1986 Aug;103(2):493–507. doi: 10.1083/jcb.103.2.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. 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]
  34. van den Pol A. N., Gorcs T. Glycine and glycine receptor immunoreactivity in brain and spinal cord. J Neurosci. 1988 Feb;8(2):472–492. doi: 10.1523/JNEUROSCI.08-02-00472.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES