Summary
A glyoxylic acid method using cryostat sections to demonstrate catecholaminergic fibres of the central nervous system was modified to show the extent of the adrenergic innervation in rat brown adipose tissue. It revealed prominent interlacing fluorescent parenchymal fibres surrounding individual adipocytes. The density of this network of fine fibres was not evident using earlier techniques. The new method also confirmed the dense networks of adrenergic fibres associated with arterial vessels. Its specificity was verified by simultaneously performing radioenzymatic determinations of tissue catecholamine levels and histochemical studies of brown adipose tissue from normal and sympathectomized rats. Chemical sympathectomy with 6-hydroxydopamine resulted in a pronounced decrease in brown adipose tissue and heart catecholamine (noradrenalin and dopamine) levels. Significantly, in brown adipose tissue of sympathectomized animals no fluorescence could be detected in terminal nerves of either the parenchyma or those of vascular smooth muscles. Nevertheless, some intense fluorescence was seen in axon bundles. The findings suggest that catecholamines of the parenchymal innervation form a larger proportion of the total catecholamine content of brown adipose tissue than was previously believed, provide stronger support for direct control of the function of multilocular adipocytes, and also confirm unpublished data reporting considerable dopamine content in brown adipose tissue.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Axelsson, S., Björklund, A., Falck, B., Lindvall, O. &Svensson, L. Å. (1973) Glyoxylic acid condensation: A new fluorescence method for the histochemical demonstration of biogenic monoamines.Acta physiol. scand. 87, 57–62.
Branard, T., Mory, G. &Nechad, M. (1980) Biogenic amines and the trophic response of brown adipose tissue InBiogenic Amines in Development (edited byParvez, H. andParvez, S.) pp. 391–439. Amsterdam: Elsevier.
Cottle, W. H., Nash, C. W., Veress, A. T., &Ferguson, B. A. (1967) Release of noradrenaline from brown fat of cold-acclimated rats.Life Sci. 6, 2267–71.
Cottle, M. K. W. &Cottle, W. H. (1969) Adrenergic fibers in brown fat.Proc. Can. Fed. Biol. Soc. 12, 94.
Cottle, M. K. W. &Cottle, W. H. (1970) Adrenergic fibers in brown fat of cold-acclimated rats.J. Histochem. Cytochem. 18, 116–9.
Cottle, M. K., Cottle, W. H., Nash, C. W. & Bickman, S. (1972) Cardiac brown fat.Proc. Int. Symp. Env. Physiol. (Bioenergetics), FASEB, pp. 122–6.
Cottle, W. H., Cottle, M. K. W. &Nash, C. W. (1975) Thermogenesis of cold-acclimated rats following 6-hydroxydopamine treatment. InTemperature Regulation and Drug Action (edited byLomax, J., Schönbaum, E. andJacob, J.) pp. 186–91. Basel: Karger.
Daniel, H. &Derry, D. M. (1969) Criteria for differentiation of brown and white fat in the rat.Can. J. Physiol. Pharmac. 47, 941–5.
de laTorre, J. C. &Surgeon, J. W. (1976) A methodological approach to rapid and sensitive monoamine histofluorescence using a modified glyoxylic acid technique: The Spg method.Histochemistry 49, 81–93.
de laTorre, J. C. (1980) Standardization of the sucrose-potassium phosphate-glyoxylic acid histofluorescence method for tissue monoamines.Neurosci. Lett. 17, 339–40.
Depocas, F., Foster, D. O., Zaror-Behrens, G., Lacelle, S. &Nadeau, B. (1984) Recovery of function in sympathetic nerves of interscapular brown adipose tissue of rats treated with 6-hydroxydopamine.Can. J. Physiol. Pharmac. 62, 1327–32.
Derry, D. M., Schönbaum, E. &Steiner, G. (1969) Two sympathetic nerve supplies to brown adipose tissue of the rat.Can. J. Physiol. Pharmac. 47, 57–63.
Falck, B., Hillarp, N. Å., Thieme, G. &Torp, A. (1962) Fluorescence of catecholamines and related compounds condensed with formaldehyde.J. Histochem. Cytochem. 10, 348–54.
Goldman, H. &Jacobowitz, D. (1971) Correlation of norepinephrine content with observations of adrenergic nerves after a single dose of 6-hydroxydopamine in the rat.J. Pharmac. exp. Ther. 176, 119–33.
Himms-Hagen, J. (1984) Brown adipose tissue thermogenesis, energy balance, and obesity.Can. J. Biochem. Cell Biol. 62, 610–7.
Jacobowitz, D. (1967) Histochemical studies of the relationship of chromaffin cells and adrenergic nerve fibers to the cardiac ganglia of several species.J. Pharmac. exp. Ther. 158, 227–40.
Kostrzewa, R. M. &Jacobowitz, D. M. (1974) Pharmacological actions of 6-hydroxydopamine.Pharmac. Rev. 26, 199–288.
Laties, A. M., Lund, R. &Jacobowitz, D. (1967) A simplified method for the histochemical localization of cardiac catecholamine-containing nerve fibers.J. Histochem. Cytochem. 15, 535–41.
Lindvall, O. &Björklund, A. (1974) The glyoxylic acid fluorescence histochemical method: A detailed account of the methodology for the visualization of central catecholamine neurons.Histochemistry 39, 97–127.
Lowry, O. H., Rosebrough, N. J., Farr, A. L., &Randall, R. J. (1951) Protein measurement with the folin phenol reagent.J. biol. Chem. 193, 265–75.
Matthews, M. R. (1980) Ultiastructural studies relevant to the possible functions of small granulecontaining cells in the rat superior cervical ganglion. InHistochemistry and Cell Biology of Autonomic Neurons, SIF Cells, and Paraneurons (edited byErankö, Ö., Soinila, S. andPäivärinta, H.) pp. 77–85. New York: Raven.
McConnell, J. (1983) Storage and long-term use of human biopsy tissue in histochemical studies.J. Neurosci. Meth. 8, 385–90.
Passon, P. G. &Peuler, J. D. (1973) A simplified radiometric assay for plasma norepinephrine and epinephrine.Analyt. Biochem. 51, 618–31.
Sorger, T. &Germinario, R. J. (1983) A direct solubilization procedure for the determination of DNA and protein in cultured fibroblast monolayers.Analyt. Biochem. 131, 254–6.
Tranzer, J. P. &Thoenen, H. (1968) An electron microscopic study of selective acute degeneration of sympathetic nerve terminals after administration of 6-hydroxydopamine.Experientia 24, 155–6.
Wirsen, C. (1964) Adrenergic innervation of adipose tissue examined by fluorescence microscopy.Nature, Lond. 202, 913.
Wirsen, C. &Hamberger, B. (1967) Catecholamines in brown fat.Nature, Lond. 214, 625–6.
Young, J. B., Saville, E., Rothwell, N. J., Stock, M. J. &Landsberg, L. (1982) Effect of diet and cold exposure on norepinephrine turnover in brown adipose tissue of the rat.J. Clin. Invest. 69, 1061–71.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Cottle, M.K.W., Cottle, W.H., Pérusse, F. et al. An improved glyoxylic acid technique for the histochemical localization of catecholamines in brown adipose tissue. Histochem J 17, 1279–1288 (1985). https://doi.org/10.1007/BF01002525
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01002525