Skip to main content
Log in

Investigations on myelinogenesis in vitro: A study of the critical period at which thyroid hormone exerts its maximum regulatory effect on the developmental experession of two myelin associated markers in cultured brain cells from embryonic mice

  • Original Articles
  • Published:
Neurochemical Research Aims and scope Submit manuscript

Abstract

Cultures of cells dissociated from embryonic mouse brain were used to assess the period in which thyroid hormone exerts its maximum influence on the regulation of the expression of two myelin associated metabolites, sulfolipids and 2′3′-cyclic nucleotide 3′-phosphohydrolase (CNP-ase). Cultures were grown for a specified number of days on a medium containing normal calf serum and then a portion were switched to a medium containing hypothyroid calf serum for 2 days. One half of these cultures were then supplemented with 50 nM triiodothyronine and growth was continued in all cultures for 3 more days. The cells were then assayed for CNP-ase activity and for their ability to incorporate35SO4 into sulfolipids. Studies with both myelin markers showed that in the earlier culture ages of 5, 8, and 11 days, thyroid hormone was able to fully restore the activities when abbed to cultures grown on hypothyroid calf-serum. In contrast, in the intermediate age range (15, 19, and 22 days) the restoration was partial, while in the higher ages, there was practically negligible restoration with T3. Since the culture system eliminates the possibility of a blood brain barrier and drastically decreases the complicity of other hormones, the lack of a myelinogeni response to thyroid hormone after a certain age must be attributed to the loss of sensitivity of the oligodendroglia to T3 possibly through genetic programming.

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

  1. Grave, G. D., (ed.) 1977. Thyroid hormones and brain development, Raven Press, New York.

    Google Scholar 

  2. Balazs, R., Brooksbank, B. W. L., Davison, A. N., Eayrs, J. T., andWilson, D. A. 1969. The effect of neonatal thyroidectomy on myelination in the rat brain. Brain Res. 15:219–232.

    Google Scholar 

  3. Balazs, R., Brooksbank, B. W. L., Patel, A. J., Johnson, A. L., andWilson, D. A. 1971. Incorporation of35[S]sulfate into brain constituents during development and the effect of thyroid hormone on myelination. Brain Res. 30:273–293.

    Google Scholar 

  4. Walravens, P., andChase, H. P. 1969 Influence of thyroid hormone on formation of myelin lipids. J. Neurochem. 16:1477–1484.

    Google Scholar 

  5. Flynn, T. J., Deshmukh, D. S., andPieringer, R. A. 1977. Effects of altered thyroid function on galactosyl diacylglycerol metabolism in myelinating rat brain. J. Biol. Chem. 252:5864–5870.

    Google Scholar 

  6. Oklund, S., andTimiras, P. S. 1977. Influences of thyroid levels in brain ontogenesis in vivo and in vitro. Pages 33–43,in Grave, G. D. (ed.), Thyroid hormones and brain development, (Vol. 1). Raven Press, New York.

    Google Scholar 

  7. Sarlieve, L. L., Fabre, M., Rebel, G., Susz, J., Vincendon, G., andMatthieu, J. M. 1983. “Myelin like” or pre-myelin structures in cultures of dissociated brain cells from 14–15 day-old embryonic mice. Pages 107–110,in Peeters, H. (ed.), Protides of the biological luids. (Vol. 30), Pergamon Press.

  8. Sarlieve, L. L., Fabre, M., Susz, J., andMatthieu, J. M. 1983. Investigations on myelination in vitro: IV. “Myelin-like” or premyelin structures in cultures of dissociated brain cells from 14–15 day-old embryonic mice. J. Neurosci. Res. 10:191–210.

    Google Scholar 

  9. Bhat, N. R. Sarlieve, L. L., Subba Rao, G., andPieringer, R. A. 1979. Investigations on myelination in vitro. Regulation by thyroid hormone in cultures of dissociated brain cells from embryonic mice. J. Biol. Chem. 254:9342–9344.

    Google Scholar 

  10. Bhat, N. R., Shanker, G., andPieringer, R. A. 1981. Investigations on myelination in vitro: Regulation of 2′, 3′-cyclic nucleotide 3′-phosphohydrolase by thyroid hormone in cultures of dissociated brain cells from embryonic mice. J. Neurochem. 37:695–701.

    Google Scholar 

  11. Bhat, N. R., Subba Rao, G., andPieringer, R. A. 1981. Investigations on myelination in vitro: Regulation of sulfolipid synthesis by thyroid hormone in cultures of dissociated brain cell from embryonic mice. J. Biol. Chem. 256:1167–1171.

    Google Scholar 

  12. Shanker, G., andPieringer, R. A. 1983. Effect of thyroid hormone on the synthesis of sialosylgalactosylceramide (GM4) in myelinogenic cultures of cells dissociated from embryonic mouse brain. Dev. Brain Res. 6:169–174.

    Google Scholar 

  13. Shanker, G., Subba Rao, G. andPieringer, R. A. 1982. Regulation of 5′-nucleotidase in dissociated brain cells of embryonic mice. Trans. Am. Soc. Neurochem. 13:134.

    Google Scholar 

  14. Shanker, G., Subba Rao, G., andPieringer, R. A. 1984. Investigations on myelinogenesis in vitro: Regulation of 5′-nucleotidase activity by thyroid hormone in cultures of dissociated cells from embryonic mouse brain. J. Neuroscience Res. 11:263–270.

    Google Scholar 

  15. Amur, S. G., Shanker, G., andPieringer, R. A. 1983. Regulation of protein methylase-I by thyroid hormone in primary cultures of dissociated brain cells from embryonic mice. J. Neurochem. 41:(supplement), S128-C.

    Google Scholar 

  16. Amur, S. G., Shanker, G., andPieringer, R. A. 1984. Regulation of myelin basic protein (arginine) methyltransferase by thyroid hormone in myelinogenic cultures of cells dissociated from embryonic mouse brain. J. Neurochem. 43:494–498.

    Google Scholar 

  17. Shanker, G., Amur, S. G., andPieringer, R. A. 1984. Critical period of thyroid hormone dependency in myelinogenic cultures. Trans. Am. Soc. Neurochem. 15:319.

    Google Scholar 

  18. Sarlieve, L. L., Campbell, G. L., Subba Rao, G., andPieringer, R. A. 1980. Investigations on myelination in vitro: Biochemical and morphological changes in cultures of dissociated brain cells from embryonic mice. Brain Res. 189:79–90.

    Google Scholar 

  19. Shanker, G., Bhat, N. R., andPieringer, R. A. 1981. Investigations on myelination in vitro: Thyroid hormone receptors in cultures of cells dissociated from embryonic mouse brain. Biosci. Rep. 1:289–297.

    Google Scholar 

  20. Bligh, E. G., andDyer, W. J. 1959 A rapid method of total lipid extraction and purification. Canad. J. Biochem. Physiol. 37:911–917.

    Google Scholar 

  21. Prohaska, J. R., Clark, D. A., andWells, W. E. 1973. Improved rapidity and precision in the determination of brain 2′,3′-cyclic nucleotide 3′-phosphohydrolase. Anal. Biochem. 56:275–282.

    Google Scholar 

  22. Lowry, O. H., Rosenbrough, N. J., Farr, A. L., andRandall, R. J. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193:265–275.

    Google Scholar 

  23. Timiras, P. S. 1972. Development and plasticity of the nervous system. Pages 129–165,in Timiras, P. S. (ed.), Developmental physiology and aging. The Macmillan Company, New York.

    Google Scholar 

  24. Sokoloff, L., andKennedy, C. 1973. The action of thyroid hormones and their influence on brain development and function. Page 295–332,in Gaull, G. E. (ed.), Biology of Brain Dysfunction. (Vol. 2) Plenum Press, New York.

    Google Scholar 

  25. Timiras, P. S., Vernadakis, A., andSherwood, N. 1968. Development and plasticity of the nervous system. Pages 261–319,in Assali N. S. (ed.), Biology of Gestation. (Vol. 2). Academic Press, New York.

    Google Scholar 

  26. Wysocki, S. J., andSegal, W. 1972. Influence of thyroid hormones on enzyme activities of myelinating rat central nervous tissue. Eur. J. Biochem. 28:183–189.

    Google Scholar 

  27. McMorris, F. A. 1983. Cyclic AMP induction of the myelin enzyme 2′,3′-cyclic nucleotide 3′-phosphohydrolase in rat oligodendrocytes. J. Neurochem. 41:506–515.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Shanker, G., Amur, S.G. & Pieringer, R.A. Investigations on myelinogenesis in vitro: A study of the critical period at which thyroid hormone exerts its maximum regulatory effect on the developmental experession of two myelin associated markers in cultured brain cells from embryonic mice. Neurochem Res 10, 617–625 (1985). https://doi.org/10.1007/BF00964401

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00964401

Keywords

Navigation