Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1975 May;72(5):1955–1959. doi: 10.1073/pnas.72.5.1955

Formation of cholinergic synapses between dissociated sympathetic neurons and skeletal myotubes of the rat in cell culture.

C A Nurse, P H O'Lague
PMCID: PMC432667  PMID: 1057775

Abstract

Sympathetic principal neurons, dissociated from superior cervical ganglia of newborn rats, were plated into cultures containing rat skeletal myotubes formed from previously plated primary myoblasts. Electrophysiological evidence is presented that the neurons developed cholinergic synapses with the myotubes. In addition, the neurons developed cholinergic synapses with each other as previously reported [O'Lague et al. (1974) Proc. Nat. Acad. Sci. USA 71, 3602-3606]. The acetylcholine receptors of myotubes differed from those of the neurons in their sensitivities to curare and hexamethonium, in a manner expected of adult muscle and ganglionic receptors. alpha-Bungarotoxin blocked synaptic transmission from neuron to myotube, but not from neuron to neuron in the same culture.

Full text

PDF
1957

Images in this article

1956Image
on p.1956

Selected References

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

  1. BROWN J. O., SATINSKY V. P. Functional restoration of the paralyzed diaphragm following the cross-union of the vagus and phrenic nerves. Am J Med Sci. 1951 Dec;222(6):613–622. doi: 10.1097/00000441-195112000-00001. [DOI] [PubMed] [Google Scholar]
  2. Bagust J., Lewis D. M., Westerman R. A. Polyneuronal innervation of kitten skeletal muscle. J Physiol. 1973 Feb;229(1):241–255. doi: 10.1113/jphysiol.1973.sp010136. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Belmar J., Eyzaguirre C. Pacemaker site of fibrillation potentials in denervated mammmalian muscle. J Neurophysiol. 1966 May;29(3):425–441. doi: 10.1152/jn.1966.29.3.425. [DOI] [PubMed] [Google Scholar]
  4. Bennett M. R., McLachlan E. M., Taylor R. S. The formation of synapses in mammalian striated muscle reinnervated with autonomic preganglionic nerves. J Physiol. 1973 Sep;233(3):501–517. doi: 10.1113/jphysiol.1973.sp010320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bennett M. R., Pettigrew A. G. The formation of synapses in striated muscle during development. J Physiol. 1974 Sep;241(2):515–545. doi: 10.1113/jphysiol.1974.sp010670. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Beránek R., Vyskocil F. The action of tubocurarine and atropine on the normal and denervated rat diaphragm. J Physiol. 1967 Jan;188(1):53–66. doi: 10.1113/jphysiol.1967.sp008123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bowman W. C., Nott M. W. Actions of sympathomimetic amines and their antagonists on skeletal muscle. Pharmacol Rev. 1969 Mar;21(1):27–72. [PubMed] [Google Scholar]
  8. Bray D. Surface movements during the growth of single explanted neurons. Proc Natl Acad Sci U S A. 1970 Apr;65(4):905–910. doi: 10.1073/pnas.65.4.905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. CHANG C. C., LEE C. Y. ISOLATION OF NEUROTOXINS FROM THE VENOM OF BUNGARUS MULTICINCTUS AND THEIR MODES OF NEUROMUSCULAR BLOCKING ACTION. Arch Int Pharmacodyn Ther. 1963 Jul 1;144:241–257. [PubMed] [Google Scholar]
  10. Cohen M. W. The development of neuromuscular connexions in the presence of D-tubocurarine. Brain Res. 1972 Jun 22;41(2):457–463. doi: 10.1016/0006-8993(72)90515-x. [DOI] [PubMed] [Google Scholar]
  11. Crain S. M. Bioelectric interactions between cultured fetal rodent spinal cord and skeletal muscle after innervation in vitro. J Exp Zool. 1970 Apr;173(4):353–369. doi: 10.1002/jez.1401730403. [DOI] [PubMed] [Google Scholar]
  12. Crain S. M., Peterson E. R. Development of neural connections in culture. Ann N Y Acad Sci. 1974 Mar 22;228(0):6–34. doi: 10.1111/j.1749-6632.1974.tb20499.x. [DOI] [PubMed] [Google Scholar]
  13. DEACOCK A. R., DAVIES T. D. The influence of certain ganglionic blocking agents on neuromuscular transmission. Br J Anaesth. 1958 May;30(5):217–225. doi: 10.1093/bja/30.5.217. [DOI] [PubMed] [Google Scholar]
  14. DEL CASTILLO J., KATZ B. On the localization of acetylcholine receptors. J Physiol. 1955 Apr 28;128(1):157–181. doi: 10.1113/jphysiol.1955.sp005297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. DIAMOND J., MILEDI R. A study of foetal and new-born rat muscle fibres. J Physiol. 1962 Aug;162:393–408. doi: 10.1113/jphysiol.1962.sp006941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Dale H. Pharmacology and Nerve-endings (Walter Ernest Dixon Memorial Lecture): (Section of Therapeutics and Pharmacology). Proc R Soc Med. 1935 Jan;28(3):319–332. [PMC free article] [PubMed] [Google Scholar]
  17. De Champlain J., Malmfors T., Olson L., Sachs C. Ontogenesis of peripheral adrenergic neurons in the rat: pre- and postnatal observations. Acta Physiol Scand. 1970 Oct;80(2):276–288. doi: 10.1111/j.1748-1716.1970.tb04791.x. [DOI] [PubMed] [Google Scholar]
  18. FATT P., KATZ B. Spontaneous subthreshold activity at motor nerve endings. J Physiol. 1952 May;117(1):109–128. [PMC free article] [PubMed] [Google Scholar]
  19. Fambrough D., Rash J. E. Development of acetylcholine sensitivity during myogenesis. Dev Biol. 1971 Sep;26(1):55–68. doi: 10.1016/0012-1606(71)90107-2. [DOI] [PubMed] [Google Scholar]
  20. Ferry C. B., Marshall A. R. An anti-curare effect of hexamethonium at the mammalian neuromuscular junction. Br J Pharmacol. 1973 Feb;47(2):353–362. doi: 10.1111/j.1476-5381.1973.tb08333.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Fischbach G. D., Cohen S. A. The distribution of acetylcholine sensitivity over uninnervated and innervated muscle fibers grown in cell culture. Dev Biol. 1973 Mar;31(1):147–162. doi: 10.1016/0012-1606(73)90326-6. [DOI] [PubMed] [Google Scholar]
  22. Fischbach G. D. Synapse formation between dissociated nerve and muscle cells in low density cell cultures. Dev Biol. 1972 Jun;28(2):407–429. doi: 10.1016/0012-1606(72)90023-1. [DOI] [PubMed] [Google Scholar]
  23. GUTH L., FRANK K. Restoration of diaphragmatic function following vagophrenic anastomosis in the rat. Exp Neurol. 1959 Apr;1(1):1–12. doi: 10.1016/0014-4886(59)90009-3. [DOI] [PubMed] [Google Scholar]
  24. Greene L. A., Sytkowski A. J., Vogel Z., Nirenberg M. W. -Bungarotoxin used as a probe for acetylcholine receptors of cultured neurones. Nature. 1973 May 18;243(5403):163–166. doi: 10.1038/243163a0. [DOI] [PubMed] [Google Scholar]
  25. Kano M., Shimada Y. Innervation and acetylcholine sensitivity of skeletal muscle cells differentiated in vitro from chick embryo. J Cell Physiol. 1971 Oct;78(2):233–242. doi: 10.1002/jcp.1040780210. [DOI] [PubMed] [Google Scholar]
  26. Kidokoro Y., Heinemann S. Synapse formation between clonal muscle cells and rat spinal cord explants. Nature. 1974 Dec 13;252(5484):593–594. doi: 10.1038/252593a0. [DOI] [PubMed] [Google Scholar]
  27. LI C. L., SHY G. M., WELLS J. Some properties of mammalian skeletal muscle fibres with particular reference to fibrillation potentials. J Physiol. 1957 Mar 11;135(3):522–535. doi: 10.1113/jphysiol.1957.sp005727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. Landmesser L. Contractile and electrical responses of vagus-innervated frog sartorius muscles. J Physiol. 1971 Mar;213(3):707–725. doi: 10.1113/jphysiol.1971.sp009410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Langley J. N., Anderson H. K. On autogenetic regeneration in the nerves of the limbs. J Physiol. 1904 Aug 22;31(5):418–428. doi: 10.1113/jphysiol.1904.sp001045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Langley J. N., Anderson H. K. The union of different kinds of nerve fibres. J Physiol. 1904 Aug 22;31(5):365–391. doi: 10.1113/jphysiol.1904.sp001042. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Letinsky M. S. The development of nerve-muscle junctions in Rana catesbeiana tadpoles. Dev Biol. 1974 Sep;40(1):129–153. doi: 10.1016/0012-1606(74)90114-6. [DOI] [PubMed] [Google Scholar]
  33. Luco C. F., Luco J. V. Sympathetic effects on fibrillary activity of denervated striated muscles. J Neurophysiol. 1971 Nov;34(6):1066–1071. doi: 10.1152/jn.1971.34.6.1066. [DOI] [PubMed] [Google Scholar]
  34. Mains R. E., Patterson P. H. Primary cultures of dissociated sympathetic neurons. I. Establishment of long-term growth in culture and studies of differentiated properties. J Cell Biol. 1973 Nov;59(2 Pt 1):329–345. doi: 10.1083/jcb.59.2.329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Miledi R., Potter L. T. Acetylcholine receptors in muscle fibres. Nature. 1971 Oct 29;233(5322):599–603. doi: 10.1038/233599a0. [DOI] [PubMed] [Google Scholar]
  36. Moore J. W., Narahashi T. Tetrodotoxin's highly selective blockage of an ionic channel. Fed Proc. 1967 Nov-Dec;26(6):1655–1663. [PubMed] [Google Scholar]
  37. O'Lague P. H., Obata K., Claude P., Furshpan E. J., Potter D. D. Evidence for cholinergic synapses between dissociated rat sympathetic neurons in cell culture. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3602–3606. doi: 10.1073/pnas.71.9.3602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Obata K. Transmitter sensitivities of some nerve and muscle cells in culture. Brain Res. 1974 Jun 14;73(1):71–88. doi: 10.1016/0006-8993(74)91008-7. [DOI] [PubMed] [Google Scholar]
  39. Olson L., Malmfors T. Growth characteristics of adrenergic nerves in the adult rat. Fluorescence histochemical and 3H-noradrenaline uptake studies using tissue transplantations to the anterior chamber of the eye. Acta Physiol Scand Suppl. 1970;348:1–112. [PubMed] [Google Scholar]
  40. Owman C., Sjöberg N. O., Swedin G. Histochemical and chemical studies on pre- and postnatal development of the different systems of "short" and "long" adrenergic neurons in peripheral organs of the rat. Z Zellforsch Mikrosk Anat. 1971;116(3):319–341. doi: 10.1007/BF00330631. [DOI] [PubMed] [Google Scholar]
  41. Patterson P. H., Chun L. L. The influence of non-neuronal cells on catecholamine and acetylcholine synthesis and accumulation in cultures of dissociated sympathetic neurons. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3607–3610. doi: 10.1073/pnas.71.9.3607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Purves D., Sakmann B. Membrane properties underlying spontaneous activity of denervated muscle fibres. J Physiol. 1974 May;239(1):125–153. doi: 10.1113/jphysiol.1974.sp010559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Ramirez B., Luco J. V. Some physiological and biochemical features of striated muscles reinnervated by preganglionic sympathetic fibers. J Neurobiol. 1973;4(6):525–533. doi: 10.1002/neu.480040605. [DOI] [PubMed] [Google Scholar]
  44. Redfern P. A. Neuromuscular transmission in new-born rats. J Physiol. 1970 Aug;209(3):701–709. doi: 10.1113/jphysiol.1970.sp009187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Robbins N., Yonezawa T. Physiological studies during formation and development of rat neuromuscular junctions in tissue culture. J Gen Physiol. 1971 Oct;58(4):467–481. doi: 10.1085/jgp.58.4.467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Thesleff S. Trophic functions of the neuron. II. Denervation and regulation of muscle. Physiological effects of denervation of muscle. Ann N Y Acad Sci. 1974 Mar 22;228(0):89–104. doi: 10.1111/j.1749-6632.1974.tb20504.x. [DOI] [PubMed] [Google Scholar]
  47. Yaffe D. Retention of differentiation potentialities during prolonged cultivation of myogenic cells. Proc Natl Acad Sci U S A. 1968 Oct;61(2):477–483. doi: 10.1073/pnas.61.2.477. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES