Skip to main content
SpringerLink
Log in

Trophic influences of sympathetic nerves and cyclic AMP on differentiation and proliferation of isolated smooth muscle cells in culture

  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Summary

Smooth muscle cells of the newborn guinea-pig vas deferens dispersed into single cells and grown in culture maintain their differentiation for approximately 5 days before undergoing dedifferentiation and mitosis. The presence of sympathetic nerve fibres in contact with the isolated cells delays this process by 3–7 days (Chamley et al., 1974). A similar delay in dedifferentiation of vas deferens smooth muscle cells in tissue culture in the presence of sympathetic ganglion extract is described in the present report, demonstrating that the trophic effect is elicited by a chemical substance. This effect is mimicked by the presence of either a confluent layer of RKA epithelial cells, dibutyryl cyclic AMP or theophylline. A similar, but considerably weaker, effect is also obtained with spinal cord and liver extracts and noradrenaline. Acetylcholine does not show an effect. It is suggested that a trophic substance (probably not noradrenaline) from sympathetic neurons activates the adenyl cyclase system of smooth muscle cells to increase the intracellular level of cyclic AMP which in turn promotes and maintains the differentiation of the cultured smooth muscle cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Anderson, W.B., Russell, T.R., Carchman, R.A., Pastan, I.: Interrelationship between adenylate cyclase activity, adenosine 3′∶5′ cyclic monophosphate phosphodiesterase activity, adenosine 3′∶5′ cyclic monophosphate levels, and growth of cells in culture. Proc. nat. Acad. Sci. (Wash.) 70, 3802–3805 (1973)

    Google Scholar 

  • Andersson, R., Nilsson, K.: Cyclic AMP and calcium in relaxation in intestinal smooth muscle. Nature (Lond.) New Biol. 238, 119–120 (1972)

    Google Scholar 

  • Armaly, M.F.: Degeneration of ciliary muscle and iris sphincter following resection of the ciliary ganglion. Trans. Amer, ophthal. Soc. 66, 475–502 (1968)

    Google Scholar 

  • Aw, E.J., Holt, P.G., Simons, P.J.: Myogenesis in vitro. Enhancement by dibutyryl cAMP. Exp. Cell Res. 83, 436–438 (1974)

    Google Scholar 

  • Beatty, C.H., Bocek, R.M., Young, M.K.: Effect of prostaglandins and epinephrine on the level of cyclic AMP-14C of smooth muscle from the Rhesus monkey. Prostaglandins 4, 661–671 (1974)

    Google Scholar 

  • Birmingham, A.T.: Sympathetic denervation of the smooth muscle of the vas deferens. J. Physiol. (Lond.) 206, 645–661 (1970)

    Google Scholar 

  • Browning, E.T., Schwartz, J.P., Breckernridge, B.McL.: Norepinephrine-sensitive properties of C-6 astrocytoma cells. Molec. Pharmacol. 10, 162–174 (1974)

    Google Scholar 

  • Bryant, S.V., Fyfe, D., Singer, M.: The effects of denervation on the ultrastructure of young limb regenerates in the newt, Triturus. Develop. Biol. 24, 577–595 (1971)

    Google Scholar 

  • Campbell, G.R., Chamley, J.H., Burnstock, G.: Development of smooth muscle cells in tissue culture. J. Anat. (Lond.) 117, 295–312 (1974)

    Google Scholar 

  • Campbell, G.R., Uehara, Y., Mark, G., Burnstock, G.: Tine structure of smooth muscle cells grown in tissue culture. J. Cell Biol. 49, 21–34 (1971 a)

    Google Scholar 

  • Campbell, G.R., Uehara, Y., Malmfors, T., Burnstock, G.: Degeneration and regeneration of smooth muscle transplants in the anterior eye chamber. An ultrastructural study. Z. Zellforsch. 117, 155–175 (1971 b)

    Google Scholar 

  • Chamley, J.H., Campbell, G.R., Burnstock, G.: Dedifferentiation, redifferentiation and bundle formation of smooth muscle cells in tissue culture: the influence of cell number and nerve fibres. J. Embryol. exp. Morph. 32, 297–323 (1974)

    Google Scholar 

  • D'Armiento, M., Johnson, G.S., Pastan, I.: Cyclic AMP and growth of fibroblasts: effect of environmental pH. Nature (Lond.) New Biol. 242, 78–80 (1973)

    Google Scholar 

  • Drachman, D.B.: Is acetylcholine the trophic neuromuscular transmitter? Arch. Neurol. 17, 206–218 (1967)

    Google Scholar 

  • Franklin, T.J., Foster, S.J.: Leakage of cyclic AMP from human diploid fibroblasts in tissue culture. Nature (Lond.) New Biol. 246, 119–120 (1973)

    Google Scholar 

  • Froehlich, J.E., Rachmeler, M.: Inhibition of cell growth in the G1 phase by adenosine 3′∶ 5′ cyclic monophosphate. J. Cell Biol. 60, 249–257 (1974)

    Google Scholar 

  • Furmanski, P., Silverman, D.J., Lubin, M.: Expression of differentiated functions in mouse neuroblastoma mediated by dibutyryl-cyclic adenosine monophosphate. Nature (Lond.) 233, 415–514 (1971)

    Google Scholar 

  • Geffen, L.B., Hughes, C.C.: Degeneration of sympathetic nerves in vitro and development of smooth muscle supersensitivity to noradrenaline. J. Physiol. (Lond.) 221, 71–84 (1972)

    Google Scholar 

  • goss, R.J.: Principles of regeneration. New York: Academic Press 1969

    Google Scholar 

  • Huang, M., Ho, A.K.S., Daly, J.W.: Accumulation of adenosine cyclic 3′∶ 5′-monophosphate in rat cerebral cortical slices. Molec. Pharmacol. 9, 711–717 (1973)

    Google Scholar 

  • Johnson, G.S., Morgan, W.D., Pastan, I.: Regulation of cell motility by cyclic AMP. Nature (Lond.) 235, 54–56 (1972)

    Google Scholar 

  • Konigsberg, I.B.: Clonal analysis of myogenesis. Science 140, 1273- 1284 (1963)

    Google Scholar 

  • Kuo, J.F., Kuo, W.N.: Regulation of /gb-adrenergic receptor and muscarinic cholinergic receptor activation of intracellular cyclic AMP and cyclic GMP levels in rat lung slices. Biochem. biophys. Res. Commun. 55, 660–665 (1973)

    Google Scholar 

  • Lebowitz, P., Singer, M.: Neurotrophic control of protein synthesis in the regenerating limb of the newt, Triturus. Nature (Lond.) 225, 824–827 (1970)

    Google Scholar 

  • Lim, R., Mitsunobu, K., Li, W.K.P.: Maturation-stimulating effect of brain extract and dibutyryl cyclic AMP on dissociated embryonic brain cells in culture. Exp. Cell Res. 79, 243–246 (1973)

    Google Scholar 

  • Lim, B., Mitsunobu, K.: Brain cells in culture: morphological transformation by a protein. Science 185, 63–66 (1974)

    Google Scholar 

  • Luco, C.F., Luco, J.V.: Sympathetic effects on fibrillary activity of denervated striated muscles. J. Neurophysiol. 34, 1066–1071 (1971)

    Google Scholar 

  • Mendez, J., Aranda, L.C., Luco, J.V.: Antifibrillary effect of adrenergic fibres on denervated striated muscles. J. Neurophysiol. 33, 882–890 (1970)

    Google Scholar 

  • Moscowitz, J., Krishna, G.: The effect of prostaglandin E1 on norepinephrine-induced membrane depolarisation and cyclic AMP accumulation in brown fat cells. Pharmacol. 10, 129–135 (1973)

    Google Scholar 

  • Newsome, D.A., Fletcher, R.T., Robison, W.G. Jr., Kenyon, K.R., Chader, G.J.: Effects of cyclic AMP and Sephadex fractions of chick embryo extract on cloned retinal pigmented epithelium in tissue culture. J. Cell Biol. 61, 369–382 (1974)

    Google Scholar 

  • Peterson, E.R., Grain, S.M.: Regeneration and innervation in cultures of adult mammalian skeletal muscle coupled with fetal rodent spinal cord. Exp. Neurol. 36, 136–139 (1972)

    Google Scholar 

  • Pfaffman, M.A., McFarland, S.A.: The effects of theophylline and cyclic AMP on intestinal smooth muscle contractions. Europ. J. Pharmacol. 23, 147–152 (1973)

    Google Scholar 

  • Prasad, K.N., Kumar, S., Gilmer, K., Vernadakis, A.: Cyclic AMP-induced differentiated neuroblastoma cells: changes in total nucleic acid and protein contents. Biochem. biophys. Res. Commun. 50, 973–977 (1973)

    Google Scholar 

  • Prosser, C.L.: Smooth muscle. Ann. Rev. Physiol. 36, 503–535 (1974)

    Google Scholar 

  • Radius, R., Langham, M.E.: Cyclic AMP and the ocular responses to norepinephrine. Exp. Eye Res. 17, 219–229 (1973)

    Google Scholar 

  • Robison, G.A., Butcher, R.W., Sutherland, E.W.: Cyclic AMP. Ann. Rev. Biochem. 37, 149–174 (1968)

    Google Scholar 

  • Schultz, G., Hardman, J.G.: The effect of norepinephrine on cyclic nucleotide concentrations in rat ductus deferens. Arch. Pharmacol. 282 (Suppl.) R88 (1974)

    Google Scholar 

  • Shapiro, D.L.: Morphological and biochemical alterations in foetal rat brain cells cultured in the presence of monobutyryl cyclic AMP. Nature (Lond.) 241, 203–204 (1973)

    Google Scholar 

  • Smith, T.E. Jr., Berndt, W.O.: The establishment of beating myocardial cells in long-term culture in fluid medium. Exp. Cell Res. 36, 179–191 (1964)

    Google Scholar 

  • Steiner, G., Shöenbaum, E.: Immunosympathectomy, p. 253. Amsterdam: Elsevier 1972

    Google Scholar 

  • Teel, R.W., Hall, R.G.: Effect of dibutyryl cyclic AMP on the restoration of contact inhibition in tumor cells and its relationship to cell density and the cell cycle. Exp. Cell Res. 76, 390–394 (1973)

    Google Scholar 

  • Thesleff, S.: Effects of motor innervation on the chemical sensitivity of skeletal muscle. Physiol. Rev. 40, 734–752 (1960)

    Google Scholar 

  • Thoenen, H., Tranzer, J.P.: Chemical sympathectomy by selective destruction of adrenergic nerve endings with 6-hydroxydopamine. Naunyn-Schmiedebergs Arch. Pharmak. exp. Path. 261, 271–288 (1968)

    Google Scholar 

  • Tower, S.: The reaction of muscle to denervation. Physiol. Rev. 19, 1–48 (1939)

    Google Scholar 

  • Weiss, G.K., Kroker, G.F., Ratner, A.: The effect of autonomie nerve stimulation on cyclic adenosine monophosphate in skeletal muscle. Life Sci. 13, 537–543 (1973)

    Google Scholar 

  • White, N.K., Hauschka, S.D.: Muscle development in vitro. A new conditioned medium effect on colony differentiation. Exp. Cell Res. 67, 479–482 (1971)

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Zoology, University of Melbourne, 3052, Parkville, Australia

    Julie H. Chamley & Gordon R. Campbell

Authors
  1. Julie H. Chamley
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gordon R. Campbell
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

J.H. Chamley holds a fellowship and a grant-in-aid from The Life Insurance Medical Research Fund of Australia and New Zealand, and G.R. Campbell a fellowship from The National Heart Foundation of Australia. We are grateful to Miss J.D. McConnell for technical assistance and to Professor G. Burnstock for use of his laboratory facilities.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chamley, J.H., Campbell, G.R. Trophic influences of sympathetic nerves and cyclic AMP on differentiation and proliferation of isolated smooth muscle cells in culture. Cell Tissue Res. 161, 497–510 (1975). https://doi.org/10.1007/BF00224140

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00224140

Key words

Search

Navigation