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.
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Grave, G. D., (ed.) 1977. Thyroid hormones and brain development, Raven Press, New York.
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.
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.
Walravens, P., andChase, H. P. 1969 Influence of thyroid hormone on formation of myelin lipids. J. Neurochem. 16:1477–1484.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
Shanker, G., Amur, S. G., andPieringer, R. A. 1984. Critical period of thyroid hormone dependency in myelinogenic cultures. Trans. Am. Soc. Neurochem. 15:319.
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.
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.
Bligh, E. G., andDyer, W. J. 1959 A rapid method of total lipid extraction and purification. Canad. J. Biochem. Physiol. 37:911–917.
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.
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.
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.
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.
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.
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.
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.
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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
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DOI: https://doi.org/10.1007/BF00964401