Synopsis
A method for the demonstration of adrenergic nerves and enteric neurons at the same time has been developed by combining the fluorescence histochemical technique for catecholamines and the histochemical technique for NADH:Nitro BT oxidoreductase.
The method consists of a short incubation of the laminae from the wall of the intestine in an isotonic medium containing the substrate (NADH) and a tetrazolium salt (Nitro BT). After washing, the laminae are air dried, exposed to formaldehyde vapour and mounted.
The adrenergic nerves in the myenteric plexus appear brightly fluorescent on excitation with u.v. light, whereas the neurons are heavily stained by deposits of formazan. Not all the neurons of the plexus are stained, but their morphology is well preserved. Differences in staining of the neurons reflect differences in penetration of the tetrazolium salt in the tissue and into the cells. The adrenergic axons do not establish exclusive connexions with individual neurons and some isolated neurons are not associated with any adrenergic fibres.
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References
Barrnett, R. J. (1959). The demonstration with the electron microscope of the end products of histochemical reactions in relation to the fine structure of cells.Exp. Cell Res. suppl. 7, 65–89.
Björklund, A., Falck, B. &Owman, Ch. (1972). Fluorescence microscopic and microspectrofluorometric techniques for the cellular localization and characterization of biogenic amines. InThe Thyroid and Biogenic Amines (eds. J. E. Rall and I. J. Kopin), pp. 318–68. Amsterdam: North Holland.
Costa, M. &Gabella, G. (1971). Adrenergic innervation of the alimentary canal.Z. Zellforsch. mikrosk. Anat. 122, 357–77.
Furness, J. B. (1970). The origin and distribution of adrenergic fibres in the guinea-pig colon.Histochemie 27, 295–306.
Furness, J. B. & Costa, M. (1973). The adrenergic innervation of the gastrointestinal tract.Ergebn. d. Physiol. in press.
Gabella, G. (1969). Detection of nerve cells by a histochemical method.Experientia (Basel) 25, 218–19.
Gabella, G. &Costa, M. (1967). Le fibre adrenergiche nel canale alimentare.Giorn. Accad. Med. Torino 130, 1–12.
Jacobowitz, D. (1965). Histochemical studies of the autonomic innervation of the gut.J. Pharmac. exp. Ther. 149, 358–64.
Lehninger, A. L. (1951). Phosphorylation coupled to oxidation of dihydrodiphosphopyridine nucleotide.J. biol. Chem. 190, 345–59.
Norberg, K. A. (1964). Adrenergic innervation of the intestinal wall studied by fluorescence microscopy.Int. J. Neuropharmacol. 3, 379–82.
Novikoff, A. B. (1960). Biochemical and staining reactions of cytoplasmic constituents. InDeveloping Cell Systems and their Control (ed. D. Rudnick), pp. 167–203. New York: The Ronald Press.
Ohkawa, H. &Prosser, C. L. (1972a). Electrical activity in myenteric and submucous plexuses of cat intestine.Am. J. Physiol. 222, 1412–19.
Ohkawa, H. &Prosser, C. L. (1972b). Functions of neurons in enteric plexuses of cat intestine.Am. J. Physiol. 222, 1420–6.
Pearse, A. G. E. (1972).Histochemistry, Theoretical and Applied, Vol. 2. Edinburgh and London: Churchill, Livingstone.
Rosenbaum, R. M. (1964). The intracellular localization of enzymes. InHandbuch der Histochemie (eds. W. Graumann and K. Neumann), Vol. 7, part 4, pp. 1–108. Stuttgart: Gustav Fisher Verlag.
Wainio, W. W. (1970).The Mammalian Mitochondrial Respiratory Chain. New York, London: Academic Press.
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Costa, M., Furness, J.B. The simultaneous demonstration of adrenergic fibres and enteric ganglion cells. Histochem J 5, 343–349 (1973). https://doi.org/10.1007/BF01004802
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DOI: https://doi.org/10.1007/BF01004802