Skip to main content
SpringerLink
Log in

Differentiation and transdifferentiation of adrenal chromaffin cells of the guinea-pig

I. Transplants to the anterior chamber of the eye

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

Summary

Autografts of adrenal chromaffin cells from adult guinea pigs to the anterior chamber of the eye were studied electron microscopically and both histo-and biochemically 4, 8 and 12 weeks after transplantation.

Transplanted chromaffin cells resembled in many respects their in-situcounterparts: they stored varying amounts of granular vesicles, which were reduced in diameter, but displayed mostly cores of low or medium electron densities suggesting predominant storage of a secondary amine. Concomitant biochemical determinations of catecholamines (CA) and phenylethanolamine N-methyltransferase (PNMT) activity revealed a distinct reduction of total CA and PNMT activity, but no change in the proportion of adrenaline (A) to noradrenaline (NA) after 4 weeks. However, an increase of NA and almost equal amounts of A and NA were found after 8 weeks.

CA-histofluorescence and electron microscopy revealed that axon-like processes with varicosities extend from chromaffin cells and contain the large “chromaffin” vesicles (100–200 nm in diameter) in addition to small clear and dense-cored vesicles (40–80 nm in diameter). Processes of chromaffin cells grew in all directions over the host iris and were also found in close proximity to smooth muscle cells the sympathetic nerve supply of which had been cut by removing the superior cervical ganglion. Administration of 5-and 6-hydroxydopamine (OHDA) resulted in characteristic labeling and ultramorphological changes in axons, but caused alterations in chromaffin cell bodies only 8 weeks after transplantation. Transplanted chromaffin cells became reinnervated by nerve fibres that are considered to be cholinergic fibres derived from the ciliary ganglion. Transplanted chromaffin cells also exhibited synapse-like contacts with each other.

The present study shows that chromaffin cells from adult guinea pigs transplanted to the anterior chamber of the eye retain a large number of differentiated properties. Formation of axon-like processes by these cells indicates that the anterior chamber of the eye favours transdifferentiation, as does tissue culture (Unsicker et al. 1978a), eliciting a cell type that displays features of both a sympathetic neurone and a SIF-cell.

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

  • Angeletti PU, Levi-Montalcini R (1970) Sympathetic nerve cell destruction in newborn mammals by 6-hydoxydopamine. Proc Natl Acad Sci USA 65:114–121

    Google Scholar 

  • Axelrod J (1962) Purification and properties of phenylethanolamine N-methyltransferase. J Biol Chem 237:1657–1660

    Google Scholar 

  • Becker K (1972) Paraganglienzellen im Ganglion cervicale uteri der Maus. Z Zellforsch 130:249–261

    Google Scholar 

  • Black IB (1978) Sympathetic neurone development. TINS 1/4:101–104

    Google Scholar 

  • Burnstock G (1976) Do some nerve cells release more than one transmitter? Neurosci 1:239–248

    Google Scholar 

  • Burnstock G, Costa M (1975) Adrenergic neurons. Chapman and Hall, London

    Google Scholar 

  • Burnstock G, Evans B, Gannon BJ, Heath J, James V (1971) A new method for destroying adrenergic nerves in adult animals using guanethidine. Br J Pharmacol 43:2095–301

    Google Scholar 

  • Carruba M, Ceccarelli B, Clementi F, Mantegazza P (1974) Selectivity in the reinnervation of iris and adrenal medulla by superior cervical ganglion after transplantation under the kidney capsule. Brain Res 77:39–53

    Google Scholar 

  • Ceccarelli B, Clementi F, Mantegazza P (1971) Synaptic transmission in the superior cervical ganglion of the cat after reinnervation by vagus fibres. J Physiol London 216:87–98

    Google Scholar 

  • Cheah TB, Geffen LB, Jarrot B, Ostberg A (1971) Action of 6-hydroxydopamine on lamb sympathetic ganglia, vas deferens and adrenal medulla: a combined histochemical, ultrastructural and biochemic comparison with the effects of reserpine. Br J Pharmacol 42:543–557

    Google Scholar 

  • Ciaranello RD, Axelrod J (1973) Genetically controlled alteration in the rate of degradation of phenylethanolamine N-methyltransferase. J Biol Chem 248:5616–5623

    Google Scholar 

  • Ciaranello RD, Dornbusch JN, Barchas JD (1972) Regulation of adrenal phenylethanolamine N-methyltransferase activity in three inbred mouse strains. Mol Pharmacol 8:511–520

    Google Scholar 

  • Ciaranello RD, Wooten GF, Axelrod J (1976) Regulation of rat adrenal dopamine β-hydroxylase. II. Receptor interaction in the regulation of enzyme synthesis and degradation. Brain Res 113:349–362

    Google Scholar 

  • Coupland RE (1953) On the morphology and adrenaline-noradrenaline content of chromaffin tissue. J Endocrinol 9:194–203

    Google Scholar 

  • Coupland RE (1959) Synthesis and storage of presser amines in adrenal medullary grafts. J Endocrinol 18:162–164

    Google Scholar 

  • Coupland RE (1965) Electron microscopic observations on the structure of the rat adrenal medulla. J Anat 99:255–272

    Google Scholar 

  • Coupland RE (1972) The chromaffin system. In: Blaschko H and Muscholl E (eds) Handbook of Experimental Pharmacology, Vol. XXXIII. Catecholamines. Springer, Berlin Heidelberg New York, pp 16–45

    Google Scholar 

  • Coupland RE, MacDougall JDB (1965) Adrenaline formation in noradrenaline-storing chromaffin cells in vitro induced by corticosterone. J Endocrinol 36:317–324

    Google Scholar 

  • Coupland RE, Weakly BS (1968) Developing chromaffin tissue in the rabbit: an electron microscopic study. J Anat London 102:425–455

    Google Scholar 

  • Coupland RE, Weakly BS (1970) Electron microscopic observation on the adrenal medulla and extraadrenal chromaffin tissue of the postnatal rabbit. J Anat London 106:213–231

    Google Scholar 

  • Csillik B, Koelle GB (1965) Histochemistry of the adrenergic and cholinergic autonomic innervation apparatus as represented by the rat iris. Acta Histochem Jena 22:350–363

    Google Scholar 

  • Douglas WW (1975) Secretomotor control of adrenal medullary secretion: synaptic, membrane and ionic events in stimulus-secretion coupling. In: Handbook of Physiology, Section 7, Vol VI, Adrenal Gland, American Physiological Society, Washington DC, pp 367–388

    Google Scholar 

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

    Google Scholar 

  • Eränkö O, Härkönen M (1963) Histochemical demonstration of fluorogenic amines in the cytoplasm of sympathetic ganglion cells of the rat. Acta Physiol Scand 58:285–286

    Google Scholar 

  • Eränkö O, Räis↭en L (1966) Demonstration of catecholamines in adrenergic nerve fibres by fixation in aqueous formaldehyde solution and fluorescence microscopy. J Histochem Cytochem 14:690–691

    Google Scholar 

  • Eränkö L, Eränkö O (1972) Effect of 6-hydroxydopamine on the ganglion cells and the small intensely fluorescent cells in the superior cervical ganglion of the rat. Acta Physiol Scand 84:115–124

    Google Scholar 

  • Euler US v (1972) Synthesis, uptake and storage of catecholamines in adrenergic nerves. The effect of drugs. In: Blaschko H and Muscholl E (eds) Catecholamines, Hdbk of Exp Pharmacology, Vol XXXIII. Springer, Berlin Heidelberg New York

    Google Scholar 

  • Feldberg W, Minz B, Tsudzimura H (1934) The mechanism of the nervous discharge of adrenaline. J Physiol London 81:286–304

    Google Scholar 

  • Furness JB, Sobels G (1976) The ultrastructure of paraganglia associated with the inferior mesenteric ganglia in the guinea pig. Cell Tissue Res 171:123–139

    Google Scholar 

  • Gorgas K, Böck P (1976) Identification of chromaffin and enterochromaffin cells in semithin sections by means of the argentaffin reaction. Mikroskopie 32:57–63

    Google Scholar 

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

    Google Scholar 

  • Grynszpan-Winograd O (1975) Ultrastructure of the chromaffin cell. In: Handbook of Physiology, Section 7, Vol VI, Adrenal Gland. American Physiological Society, Washington DC, pp 295–308

    Google Scholar 

  • Hillarp NA (1946) Structure of the synapse and the peripheral innervation apparatus of the autonomie nervous system. Hakan Ohlsons Boktrykeri, Lund, pp 1–153

    Google Scholar 

  • Höfkelt B (1952) Noradrenaline and adrenaline in mammalian tissues. Acta Physiol Scand Suppl 92:1–134

    Google Scholar 

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

  • Jacobowitz D (1970) Catecholamine fluorescence studies of adrenergic neurons and chromaffin cells in sympathetic ganglia. Fed Proc 29:1929

    Google Scholar 

  • Jones MT, Hillhouse EW, Burden JL (1977) Dynamics and mechanics of corticosteroid feedback at the hypothalamus and anterior pituitary gland. J Endocrinol 73:405–417

    Google Scholar 

  • Kirshner N (1975) Biosynthesis of the catecholamines. In: Handbook of Physiology, Section 7, Vol VI, Adrenal Gland. American Physiological Society, Washington DC, pp 341–355

    Google Scholar 

  • Kobayashi S, Coupland RE (1977) Two populations of microvesicles in the SGC (Small Granule Chromaffin) Cells of the mouse adrenal medulla. Arch Histol 3:251–259

    Google Scholar 

  • Kvetuansky R, Weise VK, Kopin IJ (1970) Elevation of adrenal tyrosine hydroxylase and phenylethanolamine-N-methyl transferase by repeated immobilization of rats. Endocrinology 87:744–749

    Google Scholar 

  • Landis SC (1976) Rat sympathetic neurons and cardiac myocytes developing in microcultures: Correlation of the fine structure of endings with neurotransmitter function in single neurons. Proc Natl Acad Sci (USA) 73:4220–4224

    Google Scholar 

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

    Google Scholar 

  • Matthews MR, Raisman G (1969) The ultrastructure and somatic efferent synapses of small granule containing cells in the superior cervical ganglion. J Anat (London) 105:255–282

    Google Scholar 

  • McLachlan EM (1974) The formation of synapses in mammalian sympathetic ganglia reinnervated with preganglionic or somatic nerves. J Physiol London 237:217–242

    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 

  • Mustonen H, Teräväinen H (1971) Synaptic connections of the paracervical (Frankenhäuser) ganglion of the rat uterus examined with the electron microscope after division of the sympathetic and sacral parasympathetic nerves. Acta Physiol Scand 82:264–267

    Google Scholar 

  • Nishida S, Trotter F, Sears M (1969) Innervation of the chamber angle of the guinea pig eye. Exp Eye Res 8:292–296

    Google Scholar 

  • O'Lague PH, MacLeish PR, Nurse CA, Claude P, Furshpan EJ, Potter DD (1976) Physiological and morphological studies on developing sympathetic neurons in dissociated cell culture. Cold Spring Harbor Symp Quant Biol 40:399–407

    Google Scholar 

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

    Google Scholar 

  • Olson L, Malmfors T (1979) Growth characteristics of adrenergic nerves in the adult rat. Acta Physiol Scand Suppl 348:1–112

    Google Scholar 

  • Orden LS van, Burke JP, Geyer M, Lodoen FV (1970) Localization of depletion-sensitive and depletion resistant norepinephrine storage sites in autonomic ganglia. J Pharmacol Exp Ther 174:56–71

    Google Scholar 

  • Piezzi PS, Cavicchia IC (1973) Explants of rat adrenal medulla. A light and electron microscopic study. Anat Rec 175:77–86

    Google Scholar 

  • Piezzi RS, Pohorecky LA, Cavicchia JC, Galleano JP (1975) Comparative effect of insuline on explants of adrenal medullary tissue and rat adrenal medulla in situ. J Endocrinol 64:323–328

    Google Scholar 

  • Pohorecky LA, Wurtman RJ (1971) Adrenocortical control of epinephrine synthesis. Pharmacol Rev 23:1–35

    Google Scholar 

  • Pohorecky LA, Piezzi RS, Wurtman RJ (1970) Steroid induction of phenylethanolamine-N-methyltransferase in adrenomedullary explants: Independence of adrenal innervation. Endocrinol 86:1466–1468

    Google Scholar 

  • Reynolds ESL (1963) The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol 17:208–213

    Article  CAS  PubMed  Google Scholar 

  • Richardson KC, Jarret L, Finke EH (1960) Embedding in epoxy resins for ultrathin sectioning in electron microscopy. Stain Technol 35:313–323

    CAS  PubMed  Google Scholar 

  • Roffi J (1965) Influence de la corticosurrénale sur le contenue en adrénaline et noradrénaline de la surrénale, chez le foetus de rat. C R Acad Sci 260:1267

    Google Scholar 

  • Siegrist G, Dolvio M, Dunant Y, Foroglou-Kerameus C, Ribeaupierre FR de, Roulier CH (1968) Ultrastructure and function of the chromaffin cells in the superior cervical ganglion of the rat. J Ultrastruct Res 25:381–407

    Google Scholar 

  • Singh J (1964) A modification of the Masson-Hamperl method for staining of argentaffin cells. Anat Anz 115:81–82

    Google Scholar 

  • Smith AD, Winkler H (1972) Fundamental mechanisms in the release of catecholamines. In: Blaschko H and Muscholl E (eds) Catecholamines, Hdbk of Exp Pharmacology, Vol XXXIII. Springer, Berlin Heidelberg New York

    Google Scholar 

  • Snedecor GW, Cochran WG (1967) Statistical methods. Ames, Iowa State University

    Google Scholar 

  • Stärne L (1972) The synthesis, uptake and storage of catecholamines in the adrenal medulla. The effect of drugs. In: Blaschko H and E. Muscholl E (eds) Catecholamines, Hdbk of Exp Pharmacol, Vol XXXIII, Springer, Berlin Heidelberg New York

    Google Scholar 

  • Thoenen H (1972) Surgical, immunological and chemical sympathectomy. In: Blaschko H and Muscholl E (eds) Catecholamines, Hdbk of Exp Pharmacol, Vol XXXIII. Springer, Berlin Heidelberg New York

    Google Scholar 

  • Unsicker K, Chamley JH (1976) Effects of dbcAMP and theophylline on rat adrenal medulla grown in tissue culture. Histochemistry 46:197–201

    CAS  PubMed  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, Ziegler W (1980) Spontaneous and nerve growth factor induced axon outgrowth from rat and bovine adrenal chromaffin cells in culture. Society for Neuroscience, 10th Annual Meeting, Cincinatti, November 9–14

  • Unsicker K, Allan IJ, Newgreen D (1976) Extraneuronal effects of 6-hydroxydopamine and extraneuronal uptake of noradrenaline. Cell Tissue Res 173:45–69

    CAS  PubMed  Google Scholar 

  • Unsicker K, Zwarg U, Habura-Flueh O (1977a) Electron microscopic evidence for the formation of synapses and synaptoid contacts in adrenal medullary grafts. Brain Res 120:533–539

    Google Scholar 

  • Unsicker K, Polonius T, Lindmar R, Löffelholz K, Wolf U (1977b) Catecholamines and 5-hydroxytryptamine in corpuscles of Stannius of the salmonid Salmo irideus L. A study correlating electron microscopical, histochemical and chemical findings. Gen Comp Endocrinol 31:121–132

    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 USA 75:3498–3502

    Google Scholar 

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

    Google Scholar 

  • Unsicker K, Habura-Flueh O, Zwarg U (1978c) Different types of small granule-containing cells and neurons in the guinea pig adrenal medulla. Cell Tissue Res 189:109–130

    Google Scholar 

  • Unsicker K, Habura-Flueh O, Zwarg U, Tschechne B, Tschechne D (1978a) Interactions between autonomic nerves and adrenal chromaffin cells in culture and in two in vivo transplant systems. In: Coupland RE, Forssmann WG (eds) Peripheral neuroendocrine interaction. Springer, Berlin Heidelberg New York, pp 60–69

    Google Scholar 

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

    Google Scholar 

  • Watanabe H (1972) The fine structure of the ciliary ganglion of the guinea pig. Arch Histol Jap 34:261–276

    Google Scholar 

  • Weiner N (1975) Control of the biosynthesis of adrenal cactecholamines by the adrenal medulla. Handbook of Physiology, Section 7: Endocrinology, Volume VI, pp 357–366

    Google Scholar 

  • Weinshilboum R, Axelrod J (1970) Dopamine-β-hydroxylase activity in the rat after hypophysectomy. Endocrinology 87:894–899

    Google Scholar 

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

    Google Scholar 

  • Williams THW (1967) Electron microscopic evidence for an autonomic interneuron. Nature 214:309–310

    Google Scholar 

  • Williams TH, Palay SL (1969) Ultrastructure of the small neurons in the superior cervical ganglion. Brain Res 15:17–34

    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

    Google Scholar 

  • Yokota R (1973) The granule containing cell somata in the superior cervical ganglion of the rat as studied by a serial sampling method for electron microscopy. Z Zellforsch 141:331–345

    Google Scholar 

  • Ziegler W, Unsicker K (1980) Differential effects of cyclic AMP and cholera toxin on nerve growth factor induced neurite outgrowth from adrenal medullary chromaffin and pheochromocytoma cells. Brain Res (submitted)

Download references

Author information

Authors and Affiliations

  1. Department of Anatomy and Cell Biology, Philipp's University, Robert-Koch-Str. 6, D-3550, Marburg, Federal Republic of Germany

    K. Unsicker, B. Tschechne & D. Tschechne

Authors
  1. K. Unsicker
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. B. Tschechne
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. D. Tschechne
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Supported by grants from the Deutsche Forschungsgemeinschaft (Un 34/6; SFB 103)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Unsicker, K., Tschechne, B. & Tschechne, D. Differentiation and transdifferentiation of adrenal chromaffin cells of the guinea-pig. Cell Tissue Res. 215, 341–367 (1981). https://doi.org/10.1007/BF00239120

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00239120

Key words

Search

Navigation