Abstract
Cyclic AMP can profoundly influence the growth and differentiation of neuronal cells in culture. In this study, the relationship between this second messenger signal transduction pathway, cell differentiation, and the expression of a retinoid-responsive, thymosin beta-10 gene was examined. Thymosin beta-10 and cognate mRNA were expressed at high levels in actively proliferating rat B104 neuroblastoma cells cultured in medium containing 10% FCS. These cells were induced to differentiate in the presence of the cAMP analog N6, 2′-O-dibutyryladenosine 3′:5′-cyclic monophosphate (Bt2-cAMP) (1 mM) and the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX) (100 μM). Expression of thymosin beta-10 mRNA was markedly inhibited (>90% and 70%, respectively) by these compounds. Addition of sodium butyrate (NaB, 1 mM) indicated that at least part of the inhibitory actions of Bt2-cAMP were due to esterase-induced release of butyrate from this compound. Adenosine (50 μM), a metabolic precursor to endogenous cyclic AMP, also inhibited accumulation of thymosin beta-10 mRNA (to <70% of control levels). The inhibitory action of Bt2-cAMP upon thymosin beta-10 mRNA levels was time dependent; levels were inhibited by >50% 24 hours after addition of the cAMP analog and by >90% after 72 hours. Serum starvation (0.2% FCS for seven days) provoked a marked increase in neurite outgrowth; this morphological change was also accompanied by a modest inhibition of thymosin beta-10 mRNA accumulation. These findings together with previous observations imply that both cyclic AMP-dependent and retinoid-responsive mechanisms coordinate thymosin beta-10 gene expression during neuroembryogenesis.
Similar content being viewed by others
References
Abemayor E., Sidell, N. (1989). Human neuroblastoma cell lines as models for the in vitro study of neoplastic and neuronal cell differentiation. Environ. Health Perspect. 80:3–15.
Adam, A., Hochholzer, L. (1981). Ganglioneuroblastoma of the posterior mediastinum: A clinicopathologic review of 80 cases. Cancer 47:373–381.
Benjamin, L. A., McGarry, R. C., Hart, D. A. (1989). Effect of retinoic acid on human neuroblastoma: Correlation between morphological differentiation and changes in plasminogen activator and inhibitor activity. Cancer Chemother Pharmcol. 25:25–31.
Blume, A.J., Dalton, C., Sheppard H. (1973). Adenosine mediated elevation of cyclic 3′:5′-adenosine monophosphate concentrations in cultured mouse neuroblastoma cells. Proc. Natl. Acad. Sci. U.S.A. 70:3099–3104.
Bottenstein, J. E., Sato, G. H. (1979). Growth of a rat neuroblastoma cell line in serum-free supplemented medium. Proc. Natl. Acad. Sci. U.S.A. 76:514–517.
Burgoyne, R.D. (1991). The Neuronal Cytoskeleton, Wiley Press, New York.
Carisson, L., Nystrom, L-E, Sundkvist, I., Markey, F., Lindberg, U. (1977). Actin polymerization is influenced by profilin, a low molecular wieght protein in non-muscle cells. J. Mol. Biol. 115:465–483.
Cassel, D., Pfeuffer, T. (1978). Mechanism of cholera toxin action: Covalent modification of the guanylnucleotide-binding protein of the adenylate cyclase system. Proc. Natl. Acad. Sci. U.S.A. 75:2669–2673.
Chomozynski, P., Sacchi, N. (1987). Singie-step method of RNA isolation by acid guanidinium thiocyanatephenol-chloroform extraction. Anal. Biochem. 162:156–159.
Christy, B.A., Nathans, D. (1989). Functional serum response elements upstream of the growth factor-inducible gene Zif268. Mol. Cell Biol. 9:4889–4895.
Clark, R.B., Gross, R., Su, Y.F., Perkins, J.P. (1974). Regulation of adenosine 3′:5′-monophosphate control in human astrocytoma cells by adenosine and adenine nucleotides. J. Biol. Chem. 249:5296–5301.
Comb, M., Birnberg, N.C., Seasholt, A., Herbert, E., Goodman, H.M. (1986). A cyclic AMP- and phorbol ester-inducible DNA element. Nature 323:353–356.
Condon, M.R., Lysze, T., Hall, A.K. (1991). Cloning of the thymosin beta-10 gene: Characterization of its structure and nucleotide sequence. Soc. Neurosci. 17(2):457:16, p 1152 (abstract)
Drummond, G.I. (1983). Cyclic nucleotides in the nervous system. Adv. Cyclic Nucleotide Res. 15:373–494.
Erickson-Viitanen, S., Ruggieri, S., Natalini, P., Horecker, B.L. (1983). Thymosin beta-10, a new analog of thymosin beta-4 in mammalian tissues. Arch. Biochem. Biophys. 225:407–413.
Fain J.N., Malbon, C.C. (1979). Regulation of adenylate cyclase by adenosine. Mol. Cell Biochem. 25:143.
Feinburg, A.P., Vogelstein, B. (1984). A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 137: 266–268.
Friedman, D.L. (1976). Role of cyclic nucleotides in cell growth and differentiation. Physiol. Rev. 56:652–708
Goodall, G.J., Horecker, B.L. (1987). Molecular cloning of the cDNA for rat spleen thymosin beta-10 and the deduced amino acid sequence. Arch. Biochem. Biophys. 256:402–405
Gottesman, M.M., Fleischmann, R.D. (1986). The role of cAMP in regulating tumor cell growth. Cancer Surv. 5:291–308
Gottesman, M.M., Fleischmann, R.D. (1986). The role of cAMP in regulating tumor cell growth. Cancer Surv. 5:291–308.
Grillon, C., Rieger, K., Bakala, J., Schott, D., Morgat, J-L., Hannappel, E., Voelter, W., Lenfant, M. (1990) Involvement of thymosin beta-4 and endoproteinase Asp-N in the biosynthesis of the tetrapeptide Ac-SerAspLysPro, a regulator of the hematopoietic system. FEBS Lett. 274:30–34.
Hall, A. (1991a). Developmental regulation of thymosin beta-10 mRNA in the human brain. Mol. Brain Res. 9:174–177.
Hall, A. (1991b) Retinoic acid and serum modulation of thymosin beta-10 gene expression in rat neuroblastoma cells. J. Mol. Neurosci. 2:229–237.
Hall, A. (1992). Retinoids and a retinoic acid receptor modulate thymosin beta-10 gene expression in rat neuroblastoma. Cell Mol. Neurobiol. 12(2):45–58.
Hall, J.M., Cole, R.D. (1986). Mechanisms of H1 accumulation in mouse neuroblastoma cells differ with different treatments. J. Biol. Chem. 261:5168–5174.
Hall, A.K., Hempstead, J., Morgan, J.I. (1990). Thymosin beta-10 levels in developing human brain and its regulation by retinoic acid in the HTB-10 neuroblastoma. Mol. Brain Res. 8:129–135.
Hall, A.K., Chen, S.-C., Hempstead, J., Morgan, J.I. (1991). Retinoic acid regulates thymosin beta-10 levels in rat neuroblastoma cells. J. Neurochem. 56(2):462–468
Hannappel, E., Xu, G.I., Morgan, J.I., Hempstead, J., Horecker, B.L. (1982). Thymosin beta-4: Aubiquitous peptide in rat and mouse tissues. Proc. Natl. Acad. Sci. U.S.A. 79:2172–2175.
Horecker, B.L., Morgan, J.I. (1984). Ubiquitous distribution of thymosin beta-4 and related peptides in vertebrate cells and tissues. Lymphokines 9:15–35.
Lugo, D., Chen S.-C., Hall, A.K., Ziai, R., Hempstead, J., Morgan, J.I. (1991). Developmental regulation of β-thymosins in the rat central nervous system. J. Neurochem. 56(2):457–461.
Mattson, M.P., Taylor-Hunter, A., Kater, S.B. (1988). Neurite outgrowth in individual neurons of neuronal population is differentially regulated by calcium and cyclic AMP. J. Neurosci. 8(5):1704–1711.
Montminy, M.H., Bilezekjian, L.M. (1987). Binding of a nuclear protein to the cyclic AMP response element of the somatostatin gene. Nature 328:175–178.
Moss, J., Vaughan, M. (1977). Mechanism of action of choleragen: Evidence for ADP-ribosyltransferase activity with arginine as an acceptor. J. Biol. Chem. 252:2455–2457.
Murdoch, G.H., Rosenfeld, M.G., Evans, R.M. (1982). Eukaryotic transcriptional regulation and chromatin associated protein phosphorylation by cyclic AMP. Science 218:1315–1317.
Pahlman, S., Odelstad, L., Larsson, E., Grotte, G., Nilsson, K. (1981). Phenotype changes of human neuroblastoma cells in culture induced by 12-O-tetradecanoyl-phorbol-13-acetate. Int. J. Cancer 28:583–589.
Pastan, I. (1975). Cyclic AMP and the malignant transformation of cells. Advances in Metabolic Disorders. Vol. 8, Somatomedins and Some Other Factors, R. Levine, R. Luft (eds). Academic Press, New York, pp 377–383.
Pieler, C., Adolf, G.R., Swetly, P. (1981). Accumulation of histone HI during chemically induced differentiation of murine neuroblastoma cells. Eur. J. Biochem. 115:329–333.
Prasad, K.N. (1975). Differentiation of neuroblastoma cells in culture. Biol. Rev. 50:129–265.
Prasad, K.N. (1977). Role of cyclic nucleotides in the differentiation of nerve cells. Cell, Tissue and Organ Culture in Neurobiology. S. Fedoroff, L. Hertz (eds). Academic Press, New York, pp 447–483.
Reboulleau, C.P. (1986). Extracellular calcium-induced neuroblastoma differentiation: Involvement of phosphatidylinositol turnover. J. Neurochem. 46:920–930
Safer, D., Elzinga, M., Nachmias, V.T. (1991). Thymosin β4 and Fx, an actin-sequestering peptide, are indistinguishable. J. Biol. 266:4029–4032.
Sattin, A., Rall, T.W. (1970). The effect of adenosine and adenine nucleotides on the cyclic adenosine 3′:5′ monophosphate content of guinea pig cerebral slices. Mol. Pharmacol. 6:23
Schuize, I., Perez-Polo, J.R. (1982). Nerve growth factor and cyclic AMP: Opposite effects on neuroblastoma-substrate adhesion. J. Neurosci. Res. 8:393–411
Shaw, G., Kamen, R. (1986). A conserved AU sequence from 3′ untranslated region of GM-CSF mRNA mediates selective mRNA degradation. Cell 46:659–667
Slemmon, J.R., Blacher, R., Danho, W., Hempstead, J., Morgan, J. (1984). Isolation and sequencing of two cerebellum-specific proteins. Proc. Natl. Acad. Sci. U.S.A. 81:6866–6870
Spinelli, W., Sonnenfeld, K.H., Ishii, D.N. (1982). Effects of phorbol ester tumor promoters and nerve growth factor on neurite outgrowth in cultured human neuroblastoma cells. Cancer Res. 42:5067–5073
Thiele, C.J., Reynolds, C.P., Israel, M.A. (1985). Decreased expression ofN-myc precedes retinoic acid-induced morphological differentiation of human neuroblastoma. Nature 313:404–406
Thiele, C.J., Cohen, P.S., Israel, M.A. (1988). Regulation ofc-myb, expression in human neuroblastoma cells during retinoic acid-induced differentiation. Mol. Cell Biol. 8:1677–1683
Thomas, P. (1980). Hybridization of denatured RNA and smali DNA fragments transferred to nitrocellulose. Proc. Natl. Acad. Sci. U.S.A. 77:5201–5205
Triesman, R. (1987). Identification and purification of a polypeptide that binds to thec-fos serum response element. EMBO J. 6:2711–2727
Wilson, t., Treisman, R. (1988). Removal of poly(A) and consequent degradation ofc-fos mRNA facilitated by 3′ AU-rich sequences. Nature 336:396–399
Woods, W.G., Tuchman, M. (1987). Neuroblastoma: The case for screening infants in North America. Pediatrics 79:869–873
Yamamoto, K.K., Gonzalez, G.A., Biggs, W.H., III, Montminy, M.R. (1988). Phosphorylation-induced binding and transcriptional efficacy of nuclear factor. CREB Nature 334:494–498
Yu, V.-C., Hochhaus, G.H., Chang, F.-H., Richards, M.L., Bourne, H.R., Sadee, W. (1988). Differentiation of human neuroblastoma cells: Marked potentiation of prostaglandin E-stimulated accumulation of cyclic AMP by retinoic acid. J. Neurochem. 51:1892–1899
Yusta, B., Ortiz-Caro, J., Pascual, A., Aranda, A. (1988). Comparison of the effects of forskolin and dibutyryl cyclic AMP in neuroblastoma cells: Evidence that some of the actions of dibutyryl cyclic AMP are mediated by butyrate. J. Neurochem. 51:1808–1818
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hall, A.K. Influence of cyclic AMP and serum factors upon expression of a retinoid-responsive gene in neuroblastoma cells. J Mol Neurosci 3, 155–163 (1992). https://doi.org/10.1007/BF02919407
Issue Date:
DOI: https://doi.org/10.1007/BF02919407