Skip to main content
SpringerLink
Log in

Embryonic rat adrenal glands in organ culture: Effects of dexamethasone, nerve growth factor and its antibodies on pheochromoblast differentiation

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

Summary

In the present study we sought to determine the developmental potentialities and restrictions of adrenal medullary cells (pheochromoblasts) by investigating their morphological and biochemical response to nerve growth factor (NGF), anti-NGF antibodies and dexamethasone (DEX) after explantation into culture at different embryonic stages. With the exception of explants taken at embryonic day 15 (E 15) cultures of embryonic adrenal glands showed neurite outgrowth, which was not influenced by the addition of NGF, anti-NGF antibodies or DEX to the culture medium during the 4-day-culture period.

Pheochromoblasts in E 17+4 explants showed spontaneous ultramorphological and biochemical maturation in terms of an increase in the number of catecholamine storage vesicles (CSVs) per μm2 of cytoplasmic area, diameters of the cores of CSVs, percentages of electron-lucent cores of CSVs indicative of increased storage of adrenaline, overall catecholamine (CA) content and relative amount of adrenaline. NGF did not significantly affect this maturational process. Anti-NGF antibodies slightly decreased the proportion of adrenaline. The most pronounced maturation was seen in response to DEX and DEX plus NGF, although a maturational state equivalent to the E 21 stage was not achieved.

E 21+4 explants showed neither spontaneous nor drug-induced biochemical maturation. Medullary cells in NGF-treated E 21 explants frequently retained the morphological features of pheochromoblasts. Treatment with anti-NGF antibodies significantly reduced the portion of adrenaline as compared to any other treatment. We conclude that under the culture conditions employed (1) a few pheochromoblasts spontaneously express a neuronal phenotype, (2) differentiation of pheochromoblasts towards chromaffin cells is enhanced by glucocorticoids but not by NGF, and (3) anti-NGF antibodies do not impair spontaneous neuritic growth and morphological maturation of pheochromoblasts, but cause a small reduction in the relative amount of adrenaline.

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 in sympathetic neurons: Effect of antiserum to nerve growth factor. Proc Natl Acad Sci 76:1246–1250

    Google Scholar 

  • Aloe L, Levi-Montalcini R (1980) Comparative studies on the effects elicited by pre- and postnatal injections of anti-NGF, guanethidine, and 6-hydroxydopamine in chromaffin and ganglion cells of the adrenal medulla and carotid body in infant rats. In: O. Eränkö, S. Soinila, H. Päivärinta (eds), Histochemistry and cell biology of autonomic neurons, SIF cells and paraneurons, Raven Press, New York, pp 221–239

    Google Scholar 

  • Angeletti PU, Levi-Montalcini R, Kettler R, Thoenen H (1972) Comparative studies on the effects of nerve growth factor on sympathetic ganglia and adrenal medulla in newborn rats. Brain Res 44:197–206

    Google Scholar 

  • Bohn MC, Goldstein M, Black IB (1981) Role of glucocorticoids in expression of the adrenergic phenotype in rat embryonic adrenal gland. Dev Biol 82:1–10

    Google Scholar 

  • Bottenstein JE, Skaper SD, Varon S, Sato GH (1980) Selective survival of neurons from chick embryo sensory ganglionic dissociates utilizing serum-free supplemented medium. Exp Cell Res 125:183–190

    Google Scholar 

  • Ciment G, Weston JA (1982) Early appearance in neural crest and crest-derived cells of an antigenic determinant present in avian neurons. Dev Biol 93:355–367

    Google Scholar 

  • Ciranello RD (1977) Regulation of phenylethanolamine N-methyltransferase synthesis and degradation. In: E. Usdin, N. Weiner, MBH. Youdim (eds), Structure and function of monoamine enzymes, Dekker, New York/Basel, pp 497–526

    Google Scholar 

  • Cogffigny H, Dupouy JP (1978) The fetal adrenals of the rat: Correlations between growth, cytology and hormonal activity with and without ACTH deficiency. Gen Comp Endocrinol 34:312–322

    Google Scholar 

  • Coupland RE (1975) The natural history of the chromaffin cell. Longmans, London

    Google Scholar 

  • Coupland RE (1980) The development and fate of catecholamine secreting endocrine cells. In: H. Parvez, S. Parvez (eds) Biogenic amines in development, Elsevier/North-Holland, pp 3–28

  • De La Torre SC, Surgeon SW (1976) A methodological approach to rapid and sensitive monoamine histofluorescence using a modified glyoxylic acid technique. The SPG method. Histochemistry 49:81–93

    Google Scholar 

  • Doupe AJ, Patterson PH, Landis SC (1980) Dissociated cell culture of SIF cells. Hormone-dependence and NGF action. Soc Neurosci Abstr 6:409

    Google Scholar 

  • Ebendal T, Olson L, Seiger Å, Hedlund KO (1980) Nerve growth factors in the rat iris. Nature (London) 286:25–28

    Google Scholar 

  • Elfvin LG (1967) The development of the secretory granules in the rat adrenal medulla. J Ultrastr Res 17:45–62

    Google Scholar 

  • El-Maghraby M, Lever JD (1980) Typification and differentiation of medullary cells in the developing rat adrenal. A histochemical and electron microscopy study. J Anat (London) 131:103–120

    Google Scholar 

  • Eränkö O, Eränkö L (1980) Induction of SIF cells by hydrocortisone or human cord serum in sympathetic ganglia and their subsequent fate in vivo and in vitro. In: O. Eränkö, S. Soinila, H. Päivärinta (eds) Histochemistry and cell biology of autonomic neurons, SIF cells, and paraneurons, Raven Press, New York, pp 17–26

    Google Scholar 

  • Eränkö O, Eränkö L, Hill CE, Burnstock G (1972) Hydrocortisone-induced increase in the number of small intensely fluorescent cells and their histochemically demonstrable catecholamine content in cultures of sympathetic ganglia of a newborn rat. Histochem J 4:49–58

    Google Scholar 

  • Falck B, Owman C (1965) A detailed methodological description of the fluorescence method for the cellular demonstration of biogenic monoamines. Acta Univ Lund Sec 27

  • Fenton EL (1970) Tissue culture assay of nerve growth factor and of the specific antiserum. Exp Cell Res 59:383–392

    Google Scholar 

  • Goedert M, Otten U, Thoenen H (1978) Biochemical effects of nerve growth factor and its antibody on the vas deferens and the adrenal medulla. Neurosci Lett 8:71–76

    Google Scholar 

  • Gorin DP, Johnson EM (1979) Experimental autoimmune model of nerve growth factor deprivation: Effects on developing peripheral sympathetic and sensory neurons. Proc Natl Acad Sci USA 76:5382–5386

    Google Scholar 

  • Gorin PD, Johnson EM (1980) Effects of long-term nerve growth factor deprivation on the nervous system of the adult rat: An experimental autoimmune approach. Brain Res 198:27–42

    Article  CAS  PubMed  Google Scholar 

  • Hervonen A, Hervonen H, Rechardt L (1972) Axonal growth from the primitive sympathetic elements of human fetal adrenal medulla. Experientia 28:178–179

    Google Scholar 

  • Hopwood D (1971) The histochemistry and electron histochemistry of chromaffin tissue. Progr Histochem Cytochem 3/1

  • Landis SC, Patterson PH (1981) Neural crest cell lineages. TINS 4:172–175

    Google Scholar 

  • Le Douarin NM (1980a) The ontogeny of the neural crest in avian embryo chimaeras. Nature (London) 286:663–669

    Google Scholar 

  • Le Douarin NM (1980b) Migration and differentiation of neural crest cells. In: A.A. Moscona, A. Monroy (eds) Current topics in developmental biology, Academic Press, New York, pp 31–85

    Google Scholar 

  • Lee V, Shelanski ML, Greene LA (1977) Specific neural and adrenal medullary antigens detected by antisera to clonal PC 12 pheochromocytoma cells. Proc Natl Acad Sci USA 74:5021–5025

    Google Scholar 

  • Lee V, Shelanski ML, Greene LA (1980) Characterization of antisera raised against cultured rats sympathetic neurons. Neuroscience 5:2239–2245

    Google Scholar 

  • Levi-Montalcini, Angeletti PU (1966) Immunosympathectomy. Pharmacol Rev 18:619–628

    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. In: Proc. 11th Annual Meeting Soc Neurosci, Los Angeles, Vol 7, p 97 (abstr)

  • Manuelidis L, Manuelidis EE (1975) Synaptic boutons and neuron-like cells in isolated adrenal gland cultures. Brain Res 96:181–186

    Google Scholar 

  • Margolis FL, Roffi J, Jost A (1966) Norepinephrine methylation in fetal rat adrenals. Science 154:275–276

    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 

  • 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 4:39–52

    Google Scholar 

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

    Google Scholar 

  • Roffi J (1968) Influence des corticosurrénales sur la synthèse d'adrénaline chez le foetus et le nouveau-né de rat et de lapin. J Physiol 60:445–494

    Google Scholar 

  • Slavinski-Turley AE, Auersperg H (1978) Cultured adrenocortical cells in various states of differentiation: Electron microscopic characterization and ultrastructural response to adrenocorticotrophin. J Endocrinol 78:427–434

    Google Scholar 

  • Somogyi I, Chubb IW, Smith AD (1975) A possible structural basis for the extracellular release of acetylcholinesterase. Proc R Soc Lond 191:271–283

    Google Scholar 

  • Teitelman G, Baker H, Joh TH, Reis DJ (1979) Appearance of catecholamine-synthesizing enzymes during development of rat sympathetic nervous system: Possible role of tissue environment. Proc Natl Acad Sci USA 76:509–513

    Google Scholar 

  • Teitelman G, Joh TH, Park D, Brodsky M, New M, Reis DJ (1982) Expression of the adrenergic phenotype in cultured fetal adrenal medullary cells: role of intrinsic and extrinsic factors. Dev Biol 89:450–459

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

  • Unsicker K (1981) Cell, tissue, and organ culture in neuroanatomy. In: Ch. Heym, W.G. Forssmann (eds) Techniques in neuroanatomical research, Springer, Berlin Heidelberg New York, pp 339–382

    Google Scholar 

  • Unsicker K (1983) Cell and tissue culture studies on the sympathoadrenal system. In: L.G. Elfvin (ed) Autonomic ganglia, John Wiley & Sons Ltd, New York, pp 475–505

    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, Hofmann HD (1983) Phenotypic plasticity of cultured bovine chromaffin cells. I. Morphological changes induced by non-chromaffin cells and organ extracts, but not by mouse and bovine nerve growth factor. Dev Brain Res 7:41–52

    Google Scholar 

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

    Google Scholar 

  • Unsicker K, Tschechne B, Teschechne D (1978b) Formation of cholinergic synapses on adrenal chromaffin cells in anterior eye chamber transplants. Brain Res 152:334–340

    Google Scholar 

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

    Google Scholar 

  • Unsicker K, Millar TJ, Hofmann HD (1982) Nerve growth factor requirement of postnatal rat adrenal medullary cells in vitro for survival, aggregate formation and maintenance of extended neurites. Dev Neurosci 5:412–417

    Google Scholar 

  • Unsicker K, Vey J, Hofmann HD, Müller TH, Wilson AJ (1984) C6 glioma-conditioned medium induces neurites outgrowth and survival of rat chromaffin cells in vitro: comparison with the effects of nerve growth factor. Proc Natl Acad Sci USA 81:2242–2246

    Google Scholar 

  • Unsicker K, Skaper SD, Varon S (1985a) Neuronotrophic and neurite promoting factors: effects on early postnatal cells from rat adrenal medulla. Dev Brain Res 17:117–129

    Google Scholar 

  • Unsicker K, Skaper SD, Varon S (1985b) Developmental changes in the responses of rat chromaffin cells to neuronotrophic and neurite promoting factors Dev Biol (in press)

  • Verhofstad AAJ, Hökfelt T, Goldstein M, Steinbusch HWM, Joosten HWJ (1979) Appearance of tyrosine hydroxylase, aromatic amino-acid decarboxylase, dopamine β-hydroxylase and phenylethanolamine N-methyltransferase during the ontogenesis of the adrenal medulla. Cell Tissue Res 200:1–13

    Google Scholar 

  • Weston JA (1970) The migration and differentiation of neural crest cells. Adv Morphol 8:41–114

    Google Scholar 

  • Wurtman RJ, Axelrod J (1965) Adrenaline synthesis control by the pituitary gland and adrenal glucocorticoids. Science 150:1464–1465

    Google Scholar 

  • Ziegler W, Hofmann HD, Unsicker K (1983) Rat adrenal nonchromaffin cells contain a neurite outgrowth-promoting factor immunologically different from nerve growth factor. Dev Brain Res 7:353–357

    Google Scholar 

Download references

Author information

Author notes

  1. Thomas J. Millar

    Present address: Centre for Neuroscience and Department of Physiology, The Medical School, Flinders University of South Australia, 5042, Bedford Park, Australia

  2. Thomas H. Müller

    Present address: Department of Pharmacology, New York University School of Medicine, 10016, New York, N.Y., USA

Authors and Affiliations

  1. Department of Anatomy and Cell Biology, Philipps University, Marburg, Federal Republic of Germany

    Klaus Unsicker, Thomas J. Millar, Thomas H. Müller & Hans -D. Hofmann

Authors
  1. Klaus Unsicker
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Thomas J. Millar
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Thomas H. Müller
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hans -D. Hofmann
    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

Unsicker, K., Millar, T.J., Müller, T.H. et al. Embryonic rat adrenal glands in organ culture: Effects of dexamethasone, nerve growth factor and its antibodies on pheochromoblast differentiation. Cell Tissue Res. 241, 207–217 (1985). https://doi.org/10.1007/BF00214643

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00214643

Key words

Search

Navigation