Abstract
Nuclear receptors for the thyroid hormone triiodothyronine (T3) have been identified in vivo in brain tissues and in vitro in mouse and rat neuroblastoma and glioma cells. The present study characterizes nuclear T3 receptors in human neuroblastoma SH-SY5Y cells and compares their levels before and after differentiation. Undifferentiated cells, grown in DME/HAM F-12 medium supplemented with 10% fetal calf serum, show an abundant single type of nuclear receptor, indicated by a straight Scatchard plot, with aK d of 0.11 nmol/l. After treatment with sodium butyrate (0.5 mM for 4 days) or NGF (2 nM for 6 days), the cells showed neuronal-like patterns (extension of neurites, slowing of growth, increased tyrosine hydroxylase activity), with a decrease in the number of nuclear T3 receptors. As sodium butyrate and NGF treatments differentiate neuroblastoma SH-SY5Y cells, these data suggest a down-regulation of T3 receptors with cell maturation.
Similar content being viewed by others
References
Kitagawa S., Obata T., Hasumura S., Pastan I., and Cheng S.-Y. 1987. A cellular 3,5,3′-Triiodo-L-Thyronine binding protein from a human carcinoma cell line. J. Biol. Chem. 262:3903.
Popescu N.C., Cheng S.-Y., and Pastan I. 1988. Chromosomal localization of the gene for a human thyroid hormone-binding protein. Am. J. Human Genetics 42:560–4.
Hashizume H., Miyamoto T., Kobayashi M., Suzuki S., Ichikawa K., Yamauchi K., Ohtsuka H., and Takeda T. 1989. Cytosolic 3,5,3′-triiodo-L-thyronine (T3)-Binding protein (CTBP) regulation of nuclear T3 binding: evidence for the presence of T3-CTBP complex-binding sites in nuclei. Endocrinology 124:1678–1683.
Segal J. 1989. A rapid, extracellular effect of 3,5,3′-triiodothyronine on sugar uptake by several tissues in the rat in vivo. Evidence for a physiological role for the thyroid hormone action at the level of the plasma membrane. Endocrinology 24:2755–2764.
De Groot L.J. 1989. Thyroid hormone nuclear receptors and their role in the metabolic action of the hormone. Biochimie 71:269–277.
Glass C.K., Halloway J.M., Devary O.V., and Rosenfeld M.G. 1988. The thyroid hormone receptor binds with opposite transcriptional effects to a common sequence motif in thyroid hormone and estrogen response elements. Cell 54:313–323.
Samuels H.H., Forman B.M., Horowitz Z.D., and Ye Z.S. 1988. Regulation of gene expression by thyroid hormone. J. Clin. Invest. 81:957–967.
Ye Z.-S., Forman B.M., Aranda A., Pascual A., Park H.-Y., Casanova J., and Samuels H.H. 1988. Rat growth hormone gene expression. Both cell specific and thyroid hormone response elements are required for thyroid hormone regulation. J. Biol. Chem. 263:7821–9.
Prasad K.N. 1980. Butyric acid; a small fatty acid with diverse biological functions. Life Sci. 27:1351–1358.
Pavelic K., and Spaventi S. 1987. Nerve Growth Factor (NGF) induced differentiation of human neuroblastoma cells. Int. J. Biochem. 19:1237–1240.
Pennypacker K.R., Kuhn D.M., and Billingsley M.L. 1989. Changes in expression of tyrosine hydroxylase immunoreactivity in human SMS-KCNR neuroblastoma following retinoic acid or phorbol ester-induced differentiation. Molecular Brain Research 5, pp. 251–258.
Gonçalves E., Lakshmanan M., and Robbins J. 1989. Triiodothyronine transport into differentiated and undifferentiated mouse neuroblastoma cells (NB41A3). Endocrinology 124:pp.293–300.
Reynolds C.P., and Perez-Polo J.R. 1989. Nerve growth factor induces neurite outgrowth in a clone derived from an NGF-insensitive human neuroblastoma cell line. Int. J. Devl. Neuroscience 17:125–132.
Safaei R., and Timiras P.S. 1985. Thyroid hormone binding and regulation of adrenergic enzymes in two neuroblastoma cell lines. J. Neurochem. 45:1405–1410.
Draves D.J., and Timiras P.S. 1980. Thyroid hormone effects in neural (tumor) cell culture: differential effects on triodothyronine nuclear receptors, Na+−K+-ATPase activity and intracellular electrolyte levels, Pages 291–301in Giacobini E., Vernadakis A., and Shahar A., (eds.), Tissue culture in Neurobiology, Raven Press, New York.
Hinegardner R.T. 1971. An improved fluorometric assay for DNA. Analytical Biochemistry 39:197–201.
Waymire J.C., Bjur R., and Weiner N. 1971. Assay of tyrosine hydroxylase by coupled decarboxylation of DOPA formed from [1-14C]-l-tyrosine. Anal. Biochem. 43; pp. 588–600.
Lowry O.H., Rosebrough N.J., Farr A.L., and Randall R.J. 1951. Protein measurement with the folin phenol reagent. J. Biol. Chem. 193:265–275.
Ciccarone V., Spengler B.A., Meyer M.B., Biedler J.L., and Ross R.A. 1989. Phenotypic diversification inhuman neuroblastoma cells expression of distint neural crest lineages. Cancer Res. 49:219–225.
Ortiz-Caro J., Montiel F., Pascual A., and Aranda A. 1986. Modulation of Thyroid hormone nuclear receptors by short-chain fatty acids in glial C6 cells. J. Biol. Chem. 261:13997.
Samuels H.H., Stanley F., Casanova J., and Shao T.C. 1980. Thyroid hormone nuclear receptor levels are influenced by the acetylation of chromatin associated proteins. J. Biol. Chem. 255:2499–2508.
Margarity M., Matsokis N., and Valcana T. 1983. Characterization of nuclear triiodothyronine (T3) and tetraiodothyronine (T4) binding in developing brain tissue. Mol. Cell Endocrinol. 31:333.
Timiras P.S. 1988. Thyroid hormones and the developing brain, Pages 59–82,in Meisami E. and Timiras P.S. (eds), Handbook of human growth and developmental biology, Vol. 1: part C.
Barsano C.P., Iqbal Z., Pullen G.L., Munoz B.E., and Singh S.P. 1990. Tissue-specific differences in the compartimentalization of rat nuclear triiodothyronine receptors. Acta Endocrinologica 122:181–190.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Goya, L., Timiras, P.S. Characterization of nuclear T3 receptors in human neuroblastoma cells SH-SY5Y: Effect of differentiation with sodium butyrate and nerve growth factor. Neurochem Res 16, 113–116 (1991). https://doi.org/10.1007/BF00965697
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00965697