Chick Spinal Somatic Motoneurons in Culture

  • Bernhard H. J. Juurlink


Cultures enriched in spinal somatic motoneurons are increasingly used as a test system for the detection of factors that are trophic or toxic for motoneurons (e.g., Schnaar and Schaffner, 1981; Calof and Reichardt, 1984; Dohrmann et al., 1986, 1987; McManaman et al., 1989, 1990; Juurlink et al., 1991a). Motoneurons in such cultures are obtained from embryonic spinal cord. There are basically two approaches whereby one can obtain cultures highly enriched in spinal motoneurons. One approach takes advantage of the fact that peripheral terminals of motoneurons can take up fluorescent labels and retrogradely transport these labels to the somas. Spinal cords can then be dissociated and labeled motoneurons isolated using a fluorescence-activated cell sorter (e.g., Calof and Reichardt, 1984; Schaffner et al., 1987); this approach is labor-intensive and yields small numbers of neurons, of which only about 80% are labeled motoneurons. There are also questions concerning the effects of the tracer on the motoneurons (Smith et al., 1986). The second approach takes advantage of the fact that the buoyant density of motoneurons is significantly different from that of the other neural cell populations; here motoneurons are separated from other cell populations by centrifuging cells through a density gradient (Schnaar and Schaffner, 1981; Dohrmann et al., 1986). This approach has the advantages that, technically, it is much simpler than the fluorescence-activated sorting method and much larger numbers of motoneurons can be isolated.


Spinal Cord Dorsal Root Ganglion Pasteur Pipet Muscle Extract Embryonic Spinal Cord 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Further Reading

  1. Butler, H. and Juurlink B. H. J. (1987), An Atlas for Staging Mammalian and Chick Embryos, CRC Press, Boca Raton, FL, p. 218.Google Scholar
  2. Calof, A. L. and Reichardt, L. F. (1984), Motoneurons purified by cell sorting respond to two distinct activities in myotube-conditioned medium. Develop. Biol. 106, 194–210.PubMedCrossRefGoogle Scholar
  3. Dohrmann, U., Edgar, D., and Thoenen, H. (1987), Distinct neurotrophic factors from skeletal muscle and the central nervous system interact synergistically to support the survival of cultured embryonic spinal motor neurons. Develop. Biol. 124, 145–152.PubMedCrossRefGoogle Scholar
  4. Dohrmann, U., Edgar, D., Sendtner, M., and Thoenen, H. (1986), Muscle-derived factors that support survival and promote fiber outgrowth from embryonic chick spinal motor neurons in culture. Develop. Biol. 118, 209–221.PubMedCrossRefGoogle Scholar
  5. Hamburger, V. and Hamilton, H. L. (1951), A series of normal stages in the development of the chick embryo. J. Morph. 88, 49–92.CrossRefGoogle Scholar
  6. Juurlink, B. H. J., Munoz, D. G. and Devon, R. M. (1990), Calcitonin gene-related peptide identifies spinal motoneurons in vitro. J. Neurosci. Res. 26, 238–241.PubMedCrossRefGoogle Scholar
  7. Juurlink, B. H. J., Munoz, D. G., and Devon, R. M. (1991a), Muscle derived trophic factors promote the survival of motoneurons in vitro only when serum is present in the growth medium. Int. J. Neurosci. 58, 249–254.PubMedCrossRefGoogle Scholar
  8. Juurlink, B. H. J., Munoz, D. G., and Ang, L. C. (1991b), Motoneuron survival in vitro: effects of pyruvate, a-ketoglutarate, gangliosides and potassium. Neurosci. Lett. 133, 25–28.PubMedCrossRefGoogle Scholar
  9. McManaman, J., Crawford, F., Clark, R., Richker, J., and Fuller, F. (1989), Multiple neurotrophic factors from skeletal muscle: demonstration of effects of basic fibroblast growth factor and comparisons with the 22-kilodalton choline acetyltransferase factor. J. Neurochem. 53, 1763–1771.PubMedCrossRefGoogle Scholar
  10. McManaman, J. L., Oppenheim, R. W., Prevette, D., and Marchetti, D. (1990), Rescue of motoneurons from cell death by a purified skeletal muscle polypeptide: effects of the ChAT development factor, CDF. Neuron 4, 891–898.PubMedCrossRefGoogle Scholar
  11. Schaffner, A. E., St. John, P. A. and Barker, J. L. (1987), Fluorescence-activated cell sorting of embryonic mouse and rat motoneurons and their long term survival in vitro. J. Neurosci. 7, 3088–3104.Google Scholar
  12. Schnaar, R. L. and Schaffner, A. E. (1981), Separation of cell types from embryonic chicken and rat spinal cord: characterization of motoneuron enriched fractions. J. Neurosci. 1, 204–217.PubMedGoogle Scholar
  13. Smith, R. G., Vaca, K., McManaman, J., and Appel, S. H. (1986), Selective effects of skeletal muscle extract fractions on motoneuron development in vitro. J. Neurosci. 6, 439–447.Google Scholar

Copyright information

© Springer Science+Business Media New York 1997

Authors and Affiliations

  • Bernhard H. J. Juurlink

There are no affiliations available

Personalised recommendations