The Accumulation of Acetylcholine Receptors at Nerve—Muscle Synapses in Culture

  • Gerald D. Fischbach
  • Lorna W. Role
  • Richard I. Hume

Abstract

We are studying the formation of synapses between embryonic chick ciliary-ganglion neurons and muscle fibers maintained in cell culture. Our assumption is that certain rules that govern synapse formation can be best understood by observing the behavior of individual cells in a controlled environment over periods of time that range from seconds to days. This observation is difficult if not impossible to achieve in intact embryos. The ability to visualize and manipulate growing neurons in vitro is a major advantage in this regard. In addition, cell cultures provide convenient and relevant bioassays that might lead to an understanding of the phenomenology of synapse formation on a molecular level. In this summary report, we describe some recent experiements in our laboratory concerning the early release of acetylcholine (ACh) from presynaptic terminals and the initial accumulation of ACh receptors (AChR) in the postsynaptic muscle membrane.

Keywords

Migration Ketone Trypsin Choline Diethyl 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Anderson, M.J., and Cohen, M.W., 1977 Nerve-induced and spontaneous redistribution of acetylcholine receptors on cultured muscle cells, J. Physiol. 268: 757 – 773.PubMedGoogle Scholar
  2. Anderson, M.J., Cohen, M.W., and Zorychta, E., 1977 Effects of innervation on the distribution of acetylcholine receptors on cultured muscle cells, J. Physiol. 268: 731 – 756.PubMedGoogle Scholar
  3. Bennett, H.P.J., Hudson, A.M., McMartin, C., and Purdon, G.E., 1977, Use of ODS-S for the extraction and purification of peptides in biological samples. Application to the identification of circulating metabolites of corticotropin-(l-24)-tetracospeptide and hematostatin in vivo, Biochem. J. 168: 9 – 13.PubMedGoogle Scholar
  4. Bennett, H.P.J., Hudson, A.M., Kelly, L., McMartin, C., and Purdon, G.E., 1978, A rapid method, using octadecasilyl-silica, for the extraction of certain peptides from tissues, Biochem. J. 175: 1139 – 1141.PubMedGoogle Scholar
  5. Buc-Caron, M.-H., Nystrom, P., and Fischbach, G.D., 1983, Induction of acetylcholine receptor synthesis and aggregation: Partial purification of low-molecular-weight activity, Dev. Biol. 95: 378 – 386.PubMedCrossRefGoogle Scholar
  6. Buc-Caron, M.-H., Nystrom, P., and Fischbach, G.D., 1983, Induction of acetylcholine receptor synthesis and aggregation: Partial purification of low-molecular-weight activity, Dev. Biol. 95: 378 – 386.Google Scholar
  7. Christian, C.N., Daniels, M.P., Sugiyama, H., Vogel, Z., Jacques, L., and Nelson, P.G., 1978 A factor from neurons that increases the number of acetylcholine receptor aggregates on cultured muscle cells, Proc. Natl. Acad. Sci. U.S.A. 75: 4011 – 4015.Google Scholar
  8. Cohen, S.A., 1980 Early nerve-muscle synapses in vitrorelease transmitter over postsynaptic membrane having low ACh sensitivity, Proc. Natl. Acad. Sci. U.S.A.77: 644 - 648.PubMedCrossRefGoogle Scholar
  9. Devreotes, P.N., and Fambrough, D.M., 1975, Acetylcholine receptor turnover in membranes of developing muscle fibers, J. Cell Biol. 65: 335 – 358.PubMedCrossRefGoogle Scholar
  10. Devreotes, P.N., Gardner, J.M., and Fambrough, D.M., 1977 Kinetics of biosynthesis of acetylcholine receptor and subsequent incorporation into plasma membrane of cultured chick skeletal muscle, Cell 10:365–373.PubMedCrossRefGoogle Scholar
  11. Frank, E., and Fischbach, G.D., 1979 Early events in neuromuscular junction formation in vitro: Induction of acetylcholine receptor clusters in the postsynaptic membrane and the morphology of newly formed synapses, J. Cell Biol. 83: 143 – 158.PubMedCrossRefGoogle Scholar
  12. Hammil, O.P., Marty, A., Neher, E., Sackmann, B., and Sigworth, F.J., 1981 Improved patch-clamp techniques for high resolution current recording from cells and cell-free membrane patches, Pfluegers Arch .391:85–100.Google Scholar
  13. Hume, R.I., Role, L.W., and Fischbach, G.D., 1983 ACh release from growth cones detected with patches of ACh receptor-rich membranes, (London) Nature (in press).Google Scholar
  14. Jessell, T.M., Siegel, R.E., and Fischbach, G.D., 1979 Induction of acetylcholine receptors on cultured skeletal muscle by a factor extracted from brain and spinal cord, Proc. Natl. Acad. Sci. U.S.A.76: 5397 – 5401.PubMedCrossRefGoogle Scholar
  15. Kidokoro, Y., and Yeh, E., 1982 Initial synaptic transmission at the growth cone in Xenopusnerve-muscle cultures, Proc. Natl. Acad. Sci. U.S.A. 79: 6727 – 6731.Google Scholar
  16. Role, L.W., Hume, R.E., and Fischbach, G.D., 1983 Transmitter release and receptor aggregation at ciliary neuron-muscle synapses, Soc. Neurosci. Abstr. 9: 129.Google Scholar
  17. Schuetze, S. M., Frank, E. F., and Fischbach, G. D., 1978 Channel open time and metabolic stability of synaptic and extrasynaptic acetylcholine receptors on cultured chick myotubes, Proc. Natl. Acad. Sci. U.S.A. 75:520—523.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1984

Authors and Affiliations

  • Gerald D. Fischbach
    • 1
  • Lorna W. Role
    • 1
  • Richard I. Hume
    • 1
  1. 1.Department of Anatomy and NeurobiologyWashington University School of MedicineSt. LouisUSA

Personalised recommendations