Skip to main content
Log in

Peripheral nerve extract promotes long-term survival and neurite outgrowth in cultured spinal cord neurons

  • Published:
Cellular and Molecular Neurobiology Aims and scope Submit manuscript

Summary

  1. 1.

    The hypothesis that peripheral, skeletal muscle tissue contains a trophic factor supporting central neurons has recently been investigatedin vitro by supplementing the culture medium of spinal cord neurons with muscle extracts and fractions of extract. We extended these studies asking whether or not a trophic factor is present in peripheral nerves, the connecting link between muscle and central neurons via which factors may be translocated from muscle to neurons by the retrograde transport system.

  2. 2.

    Lumbar, 8-day-old chick spinal cords were dissociated into single cells and then cultured in the presence of peripheral nerve extract. Cytosine arabinoside was added to inhibit proliferation of nonneuronal cells.

  3. 3.

    In the presence of nerve extract, spinal cord neurons survived for more than a month, extended numerous neurites, and showed activity of choline acetyltransferase. In the absence of extract, neurons attached and survived for a few days but then died subsequently in less than 10 days.

  4. 4.

    Neurite outgrowth did not occur in the absence of extract. Withdrawal of extract from the medium of established neuronal cultures caused progressive loss of both cells and neurites. Other tissues also contained neuron supporting activity but less than that found in nerve extract.

  5. 5.

    These studies indicate that peripheral nerves contain relatively high levels of spinal cord neuron-directed trophic activity, suggesting translocation of neurotrophic factor from muscle to central target neurons. The neurotrophic factor has long-term (weeks) effects, whereas short-term (days) survival is factor independent.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Adler, R., Landa, K. B., Manthorpe, M., and Varon, S. (1979). Cholinergic neuronotrophic factors. Intraocular distribution of trophic activity for ciliary neurons.Science 2041434–1436.

    Google Scholar 

  • Barde, Y. A., Edgar, D., and Thoenen, H. (1980). Sensory neurons in culture: Changing requirements for survival factors during embryonic development.Proc. Natl. Acad. Sci. 771199–1203.

    Google Scholar 

  • Barnes, D., and Sato, G. (1980). Methods for growth of cultured cells in serum-free medium.Anal. Biochem. 102255–270.

    Google Scholar 

  • Beach, R. L., Popiela, H., and Festoff, B. W. (1983). Muscle growth promoting (neurotrophic) factor is a transferrin.FEBS Lett. 156151–156.

    Google Scholar 

  • Bornstein, M. B. (1958). Reconstituted rat-tail collagen used as a substrate for tissue cultures on coverslips in maximow slides and roller tubes.Lab. Invest. 7134–137.

    Google Scholar 

  • Buckley, P. A., and Konigsberg, I. R. (1974). Myogenic fusion and the duration of the post-mitotic gap (G1).Dev. Biol. 37193–212.

    Google Scholar 

  • Claude, P., Hawrot, E., Dunis, D. A., and Campenot, R. B. (1982). Binding, internationalization, and retrograde transport of125I-nerve growth factor in cultured rat sympathetic neurons.J. Neurosci. 2431–442.

    Google Scholar 

  • Coughlin, M. D., Bloom, E. M., and Black, I. B. (1981). Characterization of a neuronal growth factor from mouse heart-cell-conditioned medium.Dev. Biol. 8256–68.

    Google Scholar 

  • Dribin, L. B., and Barrett, J. N. (1980). Conditioned medium enhances neuritic outgrowth from rat spinal cord explants.Dev. Biol. 74184–195.

    Google Scholar 

  • Ebendal, T., Hedlund, K.-O., and Norrgren, G. (1982). Nerve growth factors in chick tissues.J. Neurosci. Res. 8153–164.

    Google Scholar 

  • Fonnum, F. (1969). Radiochemical micro assays for the determination of choline acetyltransferase and acetylcholinesterase activities.Biochem. J. 115465–472.

    Google Scholar 

  • Godfrey, E. W., Schrier, B. K., and Nelson, P. G. (1980). Source and target specificities of a conditioned medium factor that increases choline acetyltransferase activity in cultured spinal cord cells.Dev. Biol. 77403–418.

    Google Scholar 

  • Guth, L. (1974). “Trophic” functions. InThe Peripheral Nervous System (Hubbard, J. I., (Ed.), Plenum Press, New York, pp. 329–343.

    Google Scholar 

  • Gutman, E. (1976). Neurotrophic relations.Annu. Rev. Physiol. 38177–216.

    Google Scholar 

  • Ham, R. G. (1965). Clonal growth of mammalian cells in a chemically defined, synthetic medium.Proc. Natl. Acad. Sci. 53288–293

    Google Scholar 

  • Hamburger, V. (1934). The effects of wing bud extirpation on the development of the central nervous system in chick embryos.J. Exp. Zool. 68449–494.

    Google Scholar 

  • Hamburger, V., and Keefe, E. L. (1944). The effects of peripheral factors on the proliferation and differentiation in the spinal cord of chick embryos.J. Exp. Zool. 96223–242.

    Google Scholar 

  • Hamburger, V., Brunso-Bechtold, J. K., and Yip, J. W. (1981). Neuronal death in the spinal ganglia of the chick embryo and its reduction by nerve growth factor.J. Neurosci. 160–71.

    Google Scholar 

  • Helfand, S. L., Riopelle, R. J., and Wessels, N. K. (1978). Non-equivalence of conditioned medium and nerve growth factor for sympathetic, parasympathetic, and sensory neurons.Exp. Cell Res. 11339–45.

    Google Scholar 

  • Henderson, C. E., Huchet, M., and Changeux, J. -P. (1981). Neurite outgrowth from embryonic chicken spinal neurons is promoted by media conditioned by muscle cells.Proc. Natl. Acad. Sci. 782625–2629.

    Google Scholar 

  • Henderson, C. E., Huchet, M., and Changeux, J. -P. (1983). Denervation increases a neurite-promoting activity in extracts of skeletal muscle.Nature 302609–611.

    Google Scholar 

  • Hill, C. E., Hendry, I. A., and Bonyhady, R. E. (1981). Avian parasympathetic neurotrophic factors: Age related increases and lack of regional specificity.Dev. Biol. 85258–261.

    Google Scholar 

  • Kaufman, L. M., and Barrett, J. N. (1983). Serum factor supporting long-term survival of rat central neurons in culture.Science 2201394–1396.

    Google Scholar 

  • Manthorpe, M., Varon, S., and Adler, R. (1981). Neurite-promoting factor in conditioned medium from RN22 schwannoma cultures: Bioassay, fractionation, and properties.J. Neurochem. 37759–767.

    Google Scholar 

  • McKeehan, W. L., Hamilton, W. G., and Ham, R. G. (1976). Selenium is an essential trace nutrient for growth of WI-38 diploid human fibroblasts.Proc. Natl. Acad. Sci. 732023–2027.

    Google Scholar 

  • McKeehan, W. L., McKeehan, K. A., Hammond, S. L., and Ham, R. G. (1977). Improved medium for clonal growth of human diploid fibroblasts at low concentrations of serum proteins.In Vitro 13399–416.

    Google Scholar 

  • Nishi, R., and Berg, D. K. (1981). Two components from eye tissue that differentially stimulate the growth and development of ciliary ganglion neurons in cell culture.J. Neurosci. 1505–513.

    Google Scholar 

  • O'Connor, T. M., and Wyttenbach, C. R. (1974). Cell death in the embryonic chick spinal cord.J. Cell Biol. 60448–459.

    Google Scholar 

  • Patterson, P. H., and Chun, L. L. Y. (1977). The induction of acetylcholine synthesis in primary cultures of dissociated rat sympathetic neurons. I. Effects of conditioned medium.Dev. Biol. 56263–280.

    Google Scholar 

  • Popiela, H. (1978). Trophic effects of adult peripheral nerve extract on muscle cell growth and differentiationin vitro.Exp. Neurol. 62405–416.

    Google Scholar 

  • Popiela, H., Manthorpe, M., Adler, R., and Varon, S. (1978). Choline acetyltransferase-promoting activity of medium exposed to skeletal muscle cell cultures from chick embryo.Trans. Am. Soc. Neurochem. 949.

    Google Scholar 

  • Popiela, H., and Ellis, S. (1981). Neurotrophic factor: Characterization and partial purification.Dev. Biol. 83266–277.

    Google Scholar 

  • Popiela, H., Ellis, S., and Festoff, B. W. (1982). Dose-dependent initiation of myogenesis by neurotrophic factor.J. Neurosci. Res. 8547–567.

    Google Scholar 

  • Prestige, M. C. (1967). The control of cell number in the lumbar ventral horns during the development of Xenopus laevis tadpoles.J. Embryol. Exp. Morphol. 18359–387.

    Google Scholar 

  • Puck, T. T., Cieciura, S. J., and Robinson, A. J. (1958). Genetics of somatic mammalian cells. III. Long-term cultivation of euploid cells from human and animal subjects.J. Exp. Med. 108945–956.

    Google Scholar 

  • Riopelle, R. J., and Cameron, D. A. (1981). Neurite growth promoting factors of embryonic chick—ontogeny, regional distribution, and characteristics.J. Neurobiol. 12175–186.

    Google Scholar 

  • Slack, J. R., and Pockett, S. (1982). Motor neurotrophic factor in denervated adult skeletal muscle.Brain Res. 247138–140.

    Google Scholar 

  • Smith, R. G., and Appel, S. H. (1983). Extracts of skeletal muscle increases neurite outgrowth and cholinergic activity of fetal rat spinal motor neurons.Science 2191079–1081.

    Google Scholar 

  • Tanaka, H., and Obata, K. (1982). Survival and neurite growth of chick embryo spinal cord cells in serum-free cultures.Dev. Brain Res. 4313–321.

    Google Scholar 

  • Thoenen, H., Barde, Y. -A., and Edgar, D. (1981). The role of nerve growth factor (NGF) and related factors for the survival of peripheral neurons.Adv. Biochem. Psychopharmacol. 28263–273.

    Google Scholar 

  • Tuttle, J. B., Suszkiw, J. B., and Ard, M. (1980). Long-term survival and development of dissociated parasympathetic neurons in culture.Brain Res. 183161–180.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Popiela, H., Porter, T., Beach, R.L. et al. Peripheral nerve extract promotes long-term survival and neurite outgrowth in cultured spinal cord neurons. Cell Mol Neurobiol 4, 67–77 (1984). https://doi.org/10.1007/BF00710943

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00710943

Key words

Navigation