Summary
The effects of various concentrations of 6-hydroxydopamine (6OHDA) on rat adrenocortical cells in tissue culture were studied with phase contrast and electron microscopy. With 40 mg/l of 6-OHDA the first signs of alteration as revealed by microcinematography appeared in isolated cortical cells as early as 15 min after addition of the drug. There was a cessation of movement of cell organelles and an immobilisation of membrane undulations followed by the development of dark inclusion bodies. The cells underwent increasing shrinkage and collapsed by 11/2 h. Chromaffin cells were not affected until 45 min after exposure to the drug and neurons were the most resistant population. However 61/2 h after application of the drug most cells in the culture were dead. 6-OHDA applied in different doses and to adrenal expiants did not alter the sequence of events. Ultrastructurally cortex cells underwent damage along two lines: they either showed lytic changes or developed various types of dense bodies before reaching the lytic stage.
Treatment of cortical cells with 40 mg/l 5-or 6-OHDA followed by exposure to buffered 2% glyoxylic acid and heat did not produce a fluorescence within the cells. Microspectrofluorimetry on amine models with noradrenaline, 5- and 6-OHDA revealed that neither 5-nor 6-OHDA are capable to form a fluorophore with glyoxylic acid.
Similar content being viewed by others
References
Armato, V., Nussdorfer, G.G.: Tissue culture of rat adult decapsulated adrenal glands. A methodological, ultrastructural and morphometric investigation. Z. Zellforsch. 135, 245–273 (1972)
Beard, M.E.: Identification of peroxisomes in the rat adrenal cortex. J. Histochem. Cytochem. 20, 173–179 (1972)
Björklund, A., Lindvall, O., Svensson, L.A.: Mechanisms of fluorophore formation in the histochemical glyoxylic acid method for monoamines. Histochemie 32, 113–131 (1972)
Chamley, J.H., Mark, G.E., Campbell, G.R., Burnstock, G.: Sympathetic ganglia in culture. I. Neurons. Z. Zellforsch. 135, 287–314 (1972)
Ehinger, B., Falck, B.: Fluorescence microscopical demonstration of 5-hydroxydopamine in adrenergic nerves. Histochemie 18, 1–7 (1969)
Furness, J.B., Costa, M.: The use of glyoxylic acid for the fluorescence histochemical demonstration of peripheral stores of noradrenaline and 5-hydroxytryptamine in whole mounts. Histochemistry 41, 335–352 (1975)
Heikkila, R., Cohen, G.: Inhibition of biogenic amine uptake by hydrogen peroxide: A mechanism for toxic effects of 6-hydroxydopamine. Science 172, 1257–1258 (1971)
Heikkila, R., Cohen, G.: Further studies on the generation of hydrogen peroxide by 6-hydroxydopamine. Molec. Pharmacol. 8, 241–248 (1972)
Hill, C.E., Mark, G.E., Eränkö, O., Eränkö, L., Burnstock, G.: Use of tissue culture to examine the actions of guanethidine and 6-hydroxydopamine. Europ. J. Pharmacol. 23, 162–174 (1973)
Lüllmann-Rauch, R.: Lipidosis-like alterations in spinal cord and cerebellar cortex of rats treated with chlorphentermine or tricyclic antidepressants. Acta neuropath. (Berl.) 29, 237–249 (1974)
Lüllmann-Rauch, R., Scheid, D.: Intraalveolar foam cells associated with lipidosis-like alteration in lung and liver of rats treated with tricyclic psychotropic drugs. Virchows Arch. Abt. B 19, 255–268 (1975)
Magalhães, M.M., Magalhães, M.C.: Microbodies (peroxisomes) in rat adrenal cortex. J. Ultrastruct. Res. 37, 563–573 (1971)
Malmfors, T., Sachs, Ch.: Degeneration of adrenergic nerves produced by 6-hydroxydopamine. Europ. J. Pharmacol. 3, 89–92 (1968)
Sachs, Ch., Jonsson, G.: Mechanism of action of 6-hydroxydopamine. Biochem. Pharmacol.24, 1–8 (1975)
Saner, A., Thoenen, H.: Model experiments on the molecular mechanism of action of 6-hydroxydopamine. Molec. Pharmacol. 7, 147–154 (1971)
Thoenen, H.: Surgical, immunological and chemical sympathectomy. In: Handbook of experimental pharmacology, Vol. 23, Catecholamines, ed. by H. Blaschko and E. Muscholl, pp. 813–841. Berlin-Heidelberg-New York: Springer 1972
Unsicker, K.: Fine structure of the male genital tract and kidney in the Anura Xenopus laevis Daudin, Rana temporaria L. and Bufo bufo L. under normal and experimental conditions. I. Testicular interstitial tissue and seminal efferent ducts. Cell Tis., Res. 158, 215–240 (1975)
Unsicker, K., Allan, I.J., Newgreen, D.F.: Extraneuronal effects of 6-hydroxydopamine and extraneuronal uptake of noradrenaline: In-vivo and in-vitro studies on adrenocortical cells of lizards and rats. Cell Tiss. Res. 173, 45–69 (1976)
Unsicker, K., Chamley, J.H.: Effects of dbcAMP and theophylline on rat adrenal medulla grown in tissue culture. Histochemistry 46, 197–201 (1976)
Unsicker, K., Chamley, J.H.: Growth characteristics of rat adrenal medulla in tissue culture: A study correlating phase contrast, microcinematography, histochemical and electron microscopical observations. In preparation
Weglicki, W.B.: Accumulation of lipofuscine-like pigment in the rat adrenal gland as a function of vitamin-E-deficiency. J. Geront. 23, 465–475 (1968)
Author information
Authors and Affiliations
Additional information
Dedicated to Professor Berta Scharrer in honor of her 70th birthday
Supported by a grant from Deutsche Forschungsgemeinschaft (Un 34/3) and a Research Fellowship of the University of Melbourne to K.U., and a Research Fellowship and Grant-in-Aid from the Life Insurance Medical Research Fund of Australia and New Zealand to J.H.C.
Rights and permissions
About this article
Cite this article
Unsicker, K., Chamley, J.H. & McLean, J. Extraneuronal effects of 6-hydroxydopamine. Cell Tissue Res. 174, 83–97 (1976). https://doi.org/10.1007/BF00222152
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00222152