Skip to main content
SpringerLink
Log in

Appearance of tyrosine hydroxylase, aromatic amino-acid decarboxylase, dopamine β-hydroxylase and phenylethanolamine N-methyltransferase during the ontogenesis of the adrenal medulla

An immunohistochemical study in the rat

  • Published:
Cell and Tissue Research Aims and scope

Summary

The cellular localization of the enzymes tyrosine hydroxylase (TH), aromatic amino-acid decarboxylase (or dopa decarboxylase, DDC), dopamine β-hydroxylase (DBH) and phenylethanolamine N-methyltransferase (PNMT) in the adrenal medulla of adult rats and rat fetuses (14th, 17th, 18th, 19th and 21st day) was examined. In the prenatal stages the medullary blastema and an adjacent part of the primitive sympathetic trunk were also investigated. Tissues were fixed in ice-cold 4% paraformaldehyde in 0.1 M phosphate buffer (pH 7.2). Cryostat sections (10 μm in thickness) were stained by the indirect immunofluorescence technique. Rabbit antibodies to TH (isolated from human pheochromocytoma), DDC, DBH and PNMT (the latter three isolated from bovine adrenal medulla) were used. Sections incubated with serum of non-immunized rabbits were used as controls.

In the adult adrenal medulla, two cell types can be distinguished. One cell type contains only TH, DDC and DBH. The other cell type contains PNMT in addition. It is concluded that these cells correspond to the noradrenaline-(NA-) and adrenaline-(A-)storing cells respectively. In all prenatal stages TH, DDC and DBH are found in the primitive sympathetic trunk, in the medullary blastema, and in the medullary cells which have migrated into the cortical “anlage”. PNMT is observed for the first time on the 18th day. Moreover, PNMT could only be demonstrated inside the adrenal gland. From these observations it is concluded that the capacity to synthesize NA is developed even before the “medullary” cells have reached the cortical “anlage”. On the contrary, the capacity to synthesize A seems to be acquired only after this contact is established. The hypothesis is put forward that this phenomenon might indicate the induction of PNMT by glucocorticoids secreted by the fetal cortex.

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

  • Blaschko, H.: Catecholamines 1922–1971. In: Catecholamines. Handbook of experimental pharmacology. Vol. XXXIII. (H. Blaschko and E. Muscholl, eds.) pp. 1–15. Berlin: Springer 1972

    Google Scholar 

  • Chéoux, G., Roffi, J.: Dosage de l'adrénaline et de la noradrénaline dans les surrénales du foetus et du nouveau-né de rat. C.R. Acad. Sci. [D] (Paris) 260, 6702–6705 (1965)

    Google Scholar 

  • Ciaranello, R.D.: Regulation of phenylethanolamine N-methyltransferase synthesis and degradation. I. Regulation by rat adrenal glucocorticoids. Mol. Pharmacol. 14, 478–489 (1978)

    Google Scholar 

  • Ciaranello, R.D., Jacobowitz, D., Axelrod, J.: Effect of dexamethasone on phenylethanolamine N- methyltransferase in chromaffin tissue of the neonatal rat. J. Neurochem. 20, 799–805 (1973)

    Google Scholar 

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

    Google Scholar 

  • Coons, A.H.: Fluorescent antibody methods. In: General cytochemical methods. (J.F. Danielli, ed.) pp. 399–422. New York: Academic Press 1958

    Google Scholar 

  • Couchard, P., Goldstein, M., Black, IB.: Ontogenetic appearance and disappearance of tyrosine hydroxylase and catecholamines in the rat embryo. Proc. Natl. Acad. Sci. USA 74, 2986–2990 (1978)

    Google Scholar 

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

    Google Scholar 

  • Coupland, R.E.: Corticosterone and methylation of noradrenaline by extra-adrenal chromaffin tissue. J. Endocrinol. 41, 487–490 (1968)

    Google Scholar 

  • Coupland, R.E.: Observations on the form and size distribution of chromaffin granules and on the identity of adrenaline and noradrenaline-storing cells in vertebrates and man. Mem. Soc. Endocrinol. 19, 611–633 (1971)

    Google Scholar 

  • Coupland, R.E., Selby, J.E.: The blood supply of the mammalian adrenal medulla: a comparative study. J. Anat. 122, 539–551 (1976)

    Google Scholar 

  • Coupland, R.E., Kent, Ch., Kobayashi, S.: Small-granule chromaffin cells of rodent adrenal medulla: effects of insulin hypoglycaemia and amine turnover after injection of 3H-dopa. Acta Anat. 99, 336 (1977)

    Google Scholar 

  • Eränkö, O.: Cell types of the adrenal medulla. In: Ciba foundation Symposium on adrenergic mechanisms. (G.E.W. Wolstenholme and W.O. Connor, eds.) pp. 103–108. London: Churchill 1960

    Google Scholar 

  • Eränkö, O., Lempinen, M., Räisänen, L.: Adrenaline and noradrenaline in the organ of Zuckerlandl of newborn rats treated with hydrocortisone. Acta Physiol. Scand. 66, 253–254 (1966)

    Google Scholar 

  • Eränkö, O., Lempinen, M., Räisänen, L.: Effect of hydrocortisone administration in utero on the adrenaline and noradrenaline content of extra-adrenal chromaffin tissue in the rat. Acta Physiol. Scand. 69, 255–266 (1967)

    Google Scholar 

  • Freedman, L.S., Roffman, M., Goldstein, M., Fuxe, K., Hökfelt, T.: Serum and tissue dopamine-β-hydroxylase activity in hypophysectomized rats. Eur. J. Pharmacol. 24, 366–374 (1973)

    Google Scholar 

  • Fuller, R.W., Hunt, J.M.: Activity of phenethanolamine N-methyltransferase in the adrenal glands of foetal and neonatal rats. Nature 214, 190 (1967)

    Google Scholar 

  • Fuxe, K., Goldstein, M., Hökfelt, T., Joh, T.H.: Cellular localization of dopamine-β-hydroxylase and phenylethanolamine-N-methyltransferase as revealed by immunohistochemistry. In: Histochemistry of nervous transmission. Progress in brain research. Vol. 34, (O. Eränkö, ed.) pp. 127–138. Amsterdam: Elsevier 1971

    Google Scholar 

  • Geffen, L.B., Livett, B.G., Rush, R.A.: Immunohistochemical localization of protein components of catecholamine storage vesicles. J. Physiol. (Lond.) 204, 593–605 (1969)

    Google Scholar 

  • Gewirtz, G.P., Kvetňanský, R., Weise, V.K., Kopin, I.J.: Effect of hypophysectomy on adrenal dopamine β-hydroxylase activity in the rat. Mol. Pharmacol. 7, 163–168 (1971)

    Google Scholar 

  • Giannopoulos, G.: Early events in the action of glucocorticoids in developing tissues. J. Steroid Biochem. 6, 623–631 (1975)

    Google Scholar 

  • Goldstein, M., Fuxe, K., Hökfelt, T., Joh, T.H.: Immunohistochemical studies on phenylethanolamine-N-methyltransferase, dopa-decarboxylase and dopamine-β-hydroxylase. Experientia 27, 951–952 (1971)

    Google Scholar 

  • Goldstein, M., Fuxe, K., Hökfelt, T.: Characterization und tissue localization of catecholamine synthesizing enzymes. Pharmacol. Rev. 24, 293–309 (1972)

    Google Scholar 

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

    Google Scholar 

  • Grzanna, R., Coyle, J.T.: Rat adrenal dopamine-β-hydoxylase: purification and immunologic characteristics. J. Neurochem. 27, 1091–1096 (1976)

    Google Scholar 

  • Grzanna, R., Morrison, J.H., Coyle, J.T., Mollivier, M.E.: The immunohistochemical demonstration of noradrenergic neurons in the rat brain: the use of homologous antiserum to dopamine-β-hydroxylase. Neurosci. Lett. 4, 127–134 (1977)

    Google Scholar 

  • Harrison, R.G., Hoey, M.J.: The adrenal circulation. Oxford: Blackwell 1960

    Google Scholar 

  • Hartman, B.K.: Immunofluorescence of dopamine-β-hydroxylase. Application of improved methodology to the localization of the peripheral and central noradrenergic nervous system. J. Histochem. Cytochem. 21, 312–332 (1973)

    Google Scholar 

  • Hillarp, N.A., Hökfelt, B.: Evidence of adrenaline and noradrenaline in separate adrenal medullary cells. Acta Physiol. Scand. 30, 55–68 (1954)

    Google Scholar 

  • Hökfelt, B.: Noradrenaline and adrenaline in mammalian tissues. Distribution under normal and pathological conditions with special reference to the endocrine system. Acta Physiol. Scand. [Suppl.] 25, 92 (1951)

    Google Scholar 

  • Hökfelt, T., Fuxe, K., Goldstein, M., Joh, T.H.: Imniunohistochemical localization of three catecholamine synthesizing enzymes: aspects on methodology. Histochemie 33, 231–254 (1973)

    Google Scholar 

  • Hökfelt, T., Fuxe, K., Goldstein, M.: Applications of immunohistochemistry to studies on monoamine cell systems with special reference to nervous tissues. Ann. N.Y. Acad. Sci. 254, 407–432 (1975)

    Google Scholar 

  • Kamoun, A., Mialhe-Voss, C., Stutinsky, F.: Évolution de la teneur en adrénaline et noradrénaline de la surrénale foetale du rat. C.R. Acad. Sci. 259, 3079–3082 (1964)

    Google Scholar 

  • Klepac, R., Milković, K., Milković, S.: Development of steroidogenesis in the fetal rat adrenal gland: an in vitro study. J. Steroid Biochem. 8, 841–845 (1977)

    Google Scholar 

  • Laduron, P., van Gompel, P., Leysen, J., Claeys, M.: In vivo formation of epinine in adrenal medulla. Arch. Pharmacol. 286, 227–238 (1974)

    Google Scholar 

  • Lempinen, M.: Extra-adrenal chromaffin tissue of the rat and the effect of cortical hormones on it. Acta Physiol. Scand. 62, suppl. 231 (1964)

    Google Scholar 

  • Lishajko, F.: Studies on catecholamine release and uptake in adreno-medullary storage granules. Acta Physiol. Scand., [Suppl.] 362 (1971)

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

    Google Scholar 

  • Milković, S., Milković, K., Paunović, J.: The initiation of fetal adreno-corticotrophic activity in the rat. Endocrinology 92, 380–384 (1973)

    Google Scholar 

  • Moscona, A.A.: Hydrocortisone-mediated regulation of gene expression in embryonic neural retina: induction of glutamine synthetase. J. Steroid Biochem. 6, 633–638 (1975)

    Google Scholar 

  • Nagatsu, I.: Localization of dopamine-β-hydroxylase in bovine adrenal gland and rat sciatic nerves by the improved enzyme-immunocytochemical and enzyme-immunofluorescent methods. Acta Histochem. Cytochem. 7, 147–157 (1974)

    Google Scholar 

  • Nagatsu, I., Kondo, Y.: Immunoelectronmicroscopic localization of phenylethanolamine-N- methyltransferase in the bovine adrenal medulla. Histochemistry 42, 351–358 (1974)

    Google Scholar 

  • Nairn, R.C.: Fluorescent protein tracing. Edinburgh: Churchill Livingstone 1976

    Google Scholar 

  • Pankratz, D.S.: The development of the suprarenal gland in the albino rat, with consideration of its possible relation to the origin of foetal movements. Anat. Rec. 49, 31–49 (1931)

    Google Scholar 

  • Park, D.H., Goldstein, M.: Purification of tyrosine hydroxylase from pheochromocytoma tumors. Life Sci. 18, 55–60 (1976)

    Google Scholar 

  • Pease, D.C.: Buffered formaldehyde as a killing agent and primary fixative for electron microscopy. Anat. Rec. 142, 342 (1962)

    Google Scholar 

  • Pendleton, R.G., Gessner, G.: Evidence that dopamine is not a substrate for adrenal phenylethanolamine N-methyltransferase. Mol. Pharmacol. 11, 232–235 (1975)

    Google Scholar 

  • Pickel, V.M., Joh, T.H., Reis, D.J.: Monoamine-synthesizing enzymes in central dopaminergic, noradrenergic and serotonergic neurons. J. Histochem. Cytochem. 24, 792–806 (1976)

    Google Scholar 

  • Pohorecky, L.A., Wurtman, R.J.: Adrenocortical control of epinephrine synthesis. Pharmacol. Rev. 23, 1–35 (1971)

    Google Scholar 

  • Redick, JA., Thomas, J.A., Van Orden III, L.S., Van Orden, D.E., Kopin, I.J.: Immunocytochemical localization of dopamine-β-hydroxylase in adrenal chromaffin granules. Neuropharmacology 13, 1005–1014 (1974)

    Google Scholar 

  • Roffi, J.: Influence des corticosurrénales sur la synthèse d'adrénaline chez le foetus et le nouveau-né de rat et de lapin. J. Physiol. (Paris) 60, 455–494 (1968a)

    Google Scholar 

  • Roffi, J.: Évolution des quantités d'adrénaline et de noradrénaline dans les surrénales des foetus et des nouveau-nés de rat et de lapin. Ann. Endocrinol. (Paris) 29, 277–300 (1968b)

    Google Scholar 

  • Roffi, J., Margolis, F.: Synthèse d'adrénaline dans le tissue chromaffine extrasurrénalien, chez le rat nouveau-né, sous l'effect de l'hydrocortisone ou de la corticostimuline. C.R. Acad. Sci. [D] (Paris) 263, 1496–1499 (1966)

    Google Scholar 

  • Salih, E.T.Y., Coupland, R.E.: Dedifferentiation of chromaffin cells induced by steroid synthesis inhibitor metapyrone. J. Anat. 116, 433 (1978)

    Google Scholar 

  • Schümann, H.J., Brodde, O.E.: Lack of epinine formation in adrenal medulla and brain of rats during cold exposure and inhibition of dopamine β-hydroxylase. Naunyn-Schmiedebergs Arch. Pharmacol. 293, 139–144 (1976)

    Google Scholar 

  • Shepherd, D.M., West, G.B.: Noradrenaline and the suprarenal medulla. Br. J. Pharmacol. 6, 665–674 (1951)

    Google Scholar 

  • Snider, S.R., Miller, C., Prasad, A.L.N., Jackson, V., Fahn, S.: Is dopamine a neurohormone of the adrenal medulla? Studies with morphine stimulation. Naunyn-Schmiedebergs Arch. Pharmacol. 297, 17–22 (1977)

    Google Scholar 

  • Stjärne, L.: The synthesis, uptake and storage of catecholamines in the adrenal medulla. The effect of drugs. In: Catecholamines. Handbook of experimental pharmacology, Vol. XXXIII (H. Blaschko and E, Muscholl, eds.) pp. 231–269. Berlin: Springer 1972

    Google Scholar 

  • Van Orden III, L.S., Burke, J.P., Redick, J.A., Rybarczyk, K.E., Van Orden, D.E., Baker, H.A., Hartman, B.K.: Immunocytochemical evidence for particulate localization of phenylethanolamine-N-methyltransferase in adrenal medulla. Neuropharmacology 16, 129–133 (1977)

    Google Scholar 

  • West, G.B.: The comparative pharmacology of the suprarenal medulla. Q. Rev. Biol. 30, 116–137 (1955)

    Google Scholar 

  • Wurtman, R.J., Pohorecky, L.A., Baliga, B.S.: Adrenocortical control of the biosynthesis of epinephrine and proteins in the adrenal medulla. Pharmacol. Rev. 24, 411–426 (1972)

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Histology, Karolinska Institute, Stockholm, Sweden

    T. Hökfelt

  2. Department of Psychiatry, New York University Medical Center, New York, N.Y., USA

    M. Goldstein

  3. Department of Anatomy and Embryology, University of Nijmegen, Geert Grooteplein Noord 21, P.O. Box 9101, 6500, HB Nijmegen, The Netherlands

    A. A. J. Verhofstad, T. Hökfelt, M. Goldstein, H. W. M. Steinbusch & H. W. J. Joosten

Authors
  1. A. A. J. Verhofstad
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. T. Hökfelt
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. Goldstein
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. H. W. M. Steinbusch
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. H. W. J. Joosten
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

This study was supported by a grant from the Netherlands Organization for the Advancement of Pure Research (Z.W.O.) and by the Swedish Medical Research Council (04X-2887-10C). Its results have in part been reported at the 105th Meeting of the Dutch Anatomical Society (Abstract: Acta morphologica neerlando-scandinavica, 14, 251, 1976)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Verhofstad, A.A.J., Hökfelt, T., Goldstein, M. et al. 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 (1979). https://doi.org/10.1007/BF00236882

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00236882

Key words

Search

Navigation