Skip to main content
SpringerLink
Log in

Long-term effects of dexamethasone and nerve growth factor on adrenal medullary cells cultured from young adult rats

  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Summary

Normal postnatal rat chromaffin cells and rat pheochromocytoma cells are known to show extensive Nerve Growth Factor (NGF)-induced process outgrowth in culture, and this outgrowth from the postnatal chromaffin cells is abolished by the corticosteroid dexamethasone. To determine whether adult rat chromaffin cells respond to NGF and dexamethasone, dissociated adrenal medullary cells from 3-month-old rats were cultured for 30 days in the presence or absence of these agents. Such cultures contained typical chromaffin cells, chromaffin cells with processes, and neurons. Fewer than 2 % of normal adult chromaffin cells formed processes under any of the conditions studied, and statistically significant changes in this proportion were not detectable in the presence of NGF or dexamethasone. Adrenal medullary neurons, however, were observed only in the presence of NGF, in cultures with or without dexamethasone, and thus appear to be previously unreported NGF targets which require NGF for survival or process outgrowth. Dexamethasone markedly increased total catecholamine content, total content of epinephrine, and tyrosine hydroxylase activity in cultures with or without NGF. In contrast, postnatal rat chromaffin and rat pheochromocytoma cells which have been studied in culture do not produce epinephrine under any of these conditions. It is concluded that rat adrenal chromaffin cells undergo age-related changes in both structural and functional plasticity. The in vitro characteristics of rat pheochromocytoma cells more closely resemble those of postnatal than of adult rat chromaffin cells, but may not entirely reflect the properties of the majority of chromaffin cells in either age group.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Aloe L, Levi-Montalcini R (1979) Nerve growth factor-induced transformation of immature chromaffin cells in vivo into sympathetic neurons.Effect of antiserum to nerve growth factor. Proc Natl Acad Sci USA 76:1246–1250

    Google Scholar 

  • Bocchini V, Angeletti PU (1969) The nerve growth factor: purification of a 30,000 molecular weight protein. Proc Natl Acad Sci USA 64:787–794

    Google Scholar 

  • Bornstein MB (1958) Reconstituted rat-tail collagen used as a substrate for tissue cultures on coverslips in maximow slides and roller tubes. Lab Invest 7:134–137

    Google Scholar 

  • Bunge MB (1973) Fine structure of nerve fibers and growth cones of isolated sympathetic neurons in culture. J Cell Biol 56:713–775

    Google Scholar 

  • Chalfie M, Perlman RL (1976) Studies of a transplantable rat pheochromocytoma: biochemical characterization and catecholamine secretion. J Pharmacol Exp Ther 197:615–622

    Google Scholar 

  • Cohen J (1970) Approximate power and sample size determination for common one-sample and two-sample hypothesis tests. Educ Psychol Meas 30:811–831

    Google Scholar 

  • Coupland RE (1971) Observations on the form and size distribution of chromaffin granules and on identity of adrenaline and noradrenaline-storing chromaffin cells in vertebrates and man. Mem Soc Endocrinol 19:611–635

    Google Scholar 

  • Coupland RE, MacDougall JDB (1966) Adrenaline formation in noradrenalin-storing chromaffin cells in vitro induced by corticosterone. J Endocrinol 36:317–324

    Google Scholar 

  • DeLellis RA, Merk FB, Deckers P, Warren S and Balogh K (1973) Ultrastructure and in vitro growth characteristics of a transplantable rat pheochromocytoma. Cancer 32:227–235

    Google Scholar 

  • Ebendal T, Olson L, Seiger A, Hedlund K-O (1980) Nerve growth factors in the rat iris. Nature 286:25–28

    Google Scholar 

  • Edgar DH, Thoenen H (1978) Selective enzyme induction in a nerve growth factor responsive pheochromocytoma cell line (PC 12). Brain Res 154:186–190

    Google Scholar 

  • Erny RE, Berezo MW, Perlman RL (1981) Activation of tyrosine 3-monooxygenase in pheochromocytoma cells by adenosine. J Biol Chem 256:1335–1339

    Google Scholar 

  • Euler US von, Lishajko K (1966) A specific kind of noradrenaline granules in the vesicular gland and the vas deferens of the bull. Life Sci 5:687–691

    Google Scholar 

  • Fuller RW (1973) Control of epinephrine synthesis and secretion. Fed Proc 32:1772–1781

    Google Scholar 

  • Gorgas K, Böck P (1976) Morphology and histochemistry of the adrenal medulla. Various types of primary catecholamine-storing cells in the mouse adrenal medulla. Histochemistry 50:17–31

    Google Scholar 

  • Greene LA (1977) A quantitative bioassay for nerve growth factor (NGF) activity employing a clonal pheochromocytoma cell line. Brain Res 133:350–353

    Google Scholar 

  • Greene LA, Tischler AS (1976) Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc Natl Acad Sci USA 73:2424–2428

    Google Scholar 

  • Greene LA, Tischler AS (1982) PC 12 pheochromocytoma cultures in neurobiological research. In: Fedoroff S, Hertz L (eds) Advances in cellular neurobiology. Academic Press, New York, pp 373–414

    Google Scholar 

  • Grillo MA (1966) Electron microscopy of sympathetic tissues. Pharmacol Rev 18:387–399

    Google Scholar 

  • Kirshner N, Viveros OH (1972) The secretory cycle in the adrenal medulla. Pharmacol Rev 24:385–398

    Google Scholar 

  • Kobayashi S (1977) Adrenal medulla: Chromaffin cells as paraneurons. Arch Histol Jpn (Suppl) 40:61–79

    Google Scholar 

  • Lillien LE, Claude P (1981) Adrenal chromaffin cells from adult rhesus monkeys in dissociated cell culture: Response to nerve growth factor and glucocorticoids. Abst Soc Neurosci 7:286

    Google Scholar 

  • Markey KA, Kondo S, Shenkman L, Goldstein M (1980) Purification and characterization of tyrosine hydroxylase from a clonal pheochromocytoma cell line. Molec Pharmacol 17:79–85

    Google Scholar 

  • Millar TJ, Unsicker K (1981) Catecholamine-storing cells in the adrenal medulla of the pre- and postnatal rat. Acetylcholinesterase as a means for early discrimination of cell types. Cell Tissue Res 217:155–170

    Google Scholar 

  • Mueller RA, Thoenen H, Axelrod J (1970) Effect of pituitary and ACTH on the maintenance of basal tyrosine hydroxylase activity in the rat adrenal gland. Endocrinology 86:751–755

    Google Scholar 

  • Müller TH, Unsicker K (1981) High-performance liquid chromatography with electrochemical detection as a highly efficient tool for studying catecholaminergic systems.I. Quantification of noradrenaline, adrenaline and dopamine in cultured adrenal medullary cells. J Neurosci Meth 4:36–52

    Google Scholar 

  • Olson L (1970) Fluorescence histochemical evidence for axonal growth and secretion from transplanted adrenal medullary tissue. Histochemie 22:1–7

    Google Scholar 

  • Otten U, Thoenen H (1976) Selective induction of tyrosine hydroxylase and dopamine β-hydroxylase in sympathetic ganglia in organ culture: Role of glucocorticoids as modulators. Mol Pharmacol 12:353–361

    Google Scholar 

  • Otten U, Towbin M (1980) Permissive action of glucocorticoids in induction of tyrosine hydroxylase by nerve growth factor in a pheochromocytoma cell line. Brain Res 193:304–308

    Google Scholar 

  • Otten U, Schwab M, Gagnon C and Thoenen H (1977) Selective induction of tyrosine hydroxylase and dopamine β-hydroxylase by nerve growth factor: comparison between adrenal medullae and sympathetic ganglia of adult and newborn rats. Brain Res 133:291–301

    Google Scholar 

  • Owman C, Sjöberg N-O, Swedin G (1971) Histochemical and chemical studies on pre- and postnatal development of the different systems of “short” and “long” adrenergic neurons in peripheral organs of the rat. Z Zellforsch 116:319–341

    Google Scholar 

  • Petrack B, Sheppy F, Fetzer V (1968) Studies on tyrosine hydroxylase from bovine adrenal medulla. J Biol Chem 243:743–748

    Google Scholar 

  • Richards JG, Tranzer JP (1975) Localization of amine storage sites in the adrenergic cell body. A study of the superior cervical ganglion of the rat by fine structural cytochemistry. J Ultrastruct Res 53:204–216

    Google Scholar 

  • Sandquist D, Black AC Jr, Sahu Sk, Williams L, Williams TH (1979) Dexamethasone induces increased catecholamine biosynthesis in cultured neuroblastoma. Exp Cell Res 123:417–421

    Google Scholar 

  • Schubert D, LaCorbiere M, Klier FG, Steinbach JH (1980) The modulation of neurotransmitter synthesis by steroid hormones and insulin. Brain Res 190:67–79

    Google Scholar 

  • Tischler AS, Greene LA (1975) Nerve growth factor-induced process formation by cultured rat pheochromocytoma cells. Nature 258:341–342

    Google Scholar 

  • Tischler AS, Greene LA (1978) Morphologic and cytochemical properties of a clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Lab Invest 39:77–89

    Google Scholar 

  • Tischler AS, Greene LA (1980) Phenotypic plasticity of pheochromocytoma and normal adrenal medullary cells. In: Eränkö O, Soinila S, Päivärinta H (eds) Histochemistry and cell biology of autonomic neurons, SIF cells and paraneurons. Raven Press, New York, pp 61–68

    Google Scholar 

  • Tischler AS, Dichter MA, Biales B, DeLellis RA, Wolfe HJ (1976) Neuronal properties of cultured human endocrine tumor cells of proposed neural crest origin. Science 192:902–904

    Google Scholar 

  • Tischler AS, DeLellis RA, Biales B, Nunnemacher G, Carraba V, Wolfe HJ (1980) Nerve growth factor induced neurite outgrowth from normal human chromaffin cells. Lab Invest 43:399–409

    Google Scholar 

  • Unsicker K, Chamley JH (1977) Growth characteristics of postnatal rat adrenal medulla in culture. Cell Tissue Res 177:247–268

    Google Scholar 

  • Unsicker K, Krisch B, Otten U, Thoenen H (1978) Nerve growth factor-induced fiber outgrowth from isolated rat adrenal chromaffin cells: impairment by glucocorticoids. Proc Nat Acad Sci (Wash) 75:3498–3502

    Google Scholar 

  • Unsicker K, Griesser GH, Lindmar R, Löffelholz K, Wolf U (1980a) Establishment, characterization, and fibre outgrowth of isolated bovine adrenal medullary cells in long-term cultures. Neuroscience 5:1445–1460

    Google Scholar 

  • Unsicker K, Rieffert B, Ziegler W (1980b) Effects of cell culture conditions, nerve growth factor, dexamethasone and cyclic AMP on adrenal chromaffin cells in vitro. In: Eränkö O, Soinila S, Päivärinta H (eds) Histochemistry and cell biology of autonomic neurons, SIF Cells, and paraneurons. Raven Press, New York, pp 51–59

    Google Scholar 

  • Unsicker K, Tschechne B, Tschechne (1981) Differentiation and transdifferentiation of adrenal chromaffm cells of the guinea-pig.I. Transplants to the anterior chamber of the eye. Cell Tissue Res 215:341–367

    Google Scholar 

  • Vaccaro KK, Liang BT, Perelle BA, Perlman RL (1980) Tyrosine 3-monooxygenase regulates catecholamine synthesis in pheochromocytoma cells. J Biol Chem 255:6539–6541

    Google Scholar 

  • Varon S, Raiborn C (1972) Dissociation, fractionation and culture of chick embryo sympathetic ganglionic cells. J Neurocytol 1:211–221

    Google Scholar 

  • Williams LR, Sandquist D, Black AC, Williams TH (1981) Glucocorticoids increase tyrosine hydroxylase activity in cultered murine neuroblastoma. J Neurochem 36:2057–2062

    Google Scholar 

  • Wurtman RJ, Axelrod J (1966) Control of enzymatic synthesis of adrenaline in the adrenal medulla by adrenal cortical steroids. J Biol Chem 241:2301–2305

    Google Scholar 

Download references

Author information

Author notes

  1. Robert L. Perlman

    Present address: Department of Physiology and Biophysics, University of Illinois College of Medicine, Chicago, IL, USA

Authors and Affiliations

  1. Department of Pathology, Tufts University School of Medicine, Boston, USA

    Arthur S. Tischler, Gretl Nunnemacher, Gardiner M. Morse, Ronald A. DeLellis & Hubert J. Wolfe

  2. Department of Physiology, Harvard Medical School, Boston, USA

    Robert L. Perlman & Beth E. Sheard

Authors
  1. Arthur S. Tischler
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Robert L. Perlman
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Gretl Nunnemacher
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Gardiner M. Morse
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ronald A. DeLellis
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hubert J. Wolfe
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Beth E. Sheard
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tischler, A.S., Perlman, R.L., Nunnemacher, G. et al. Long-term effects of dexamethasone and nerve growth factor on adrenal medullary cells cultured from young adult rats. Cell Tissue Res. 225, 525–542 (1982). https://doi.org/10.1007/BF00214802

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00214802

Key words

Search

Navigation