Summary
This paper describes a procedure for demonstration of catecholamine-and acetylcholinesterase-containing neurons in the same section of central nervous tissue. The brains are first processed according to the glyoxylic acid (GA) fluorescence method for catecholamine neurons, i.e. perfused with an ice-cold GA solution, sectioned on a Vibratome instrument, immersed in a GA solution and dried under a stream of warm air. The unmounted sections are examined and photographed in the fluorescence microscope, and then stained for acetylcholinesterase according to Holmstedt's modification of the Koelle thiocholine method (incubation for 4–6 h with acetylthiocholine as substrate and Mipafox as inhibitor of non-specific cholinesterases). The sections are then examined in the light microscope, rephotographed, and the picture compared with that following the GA reaction. The present technique makes possible, for the first time, detailed light microscopical studies of the morphological relations between central catecholamine- and acetylcholinesterase-containing neurons in the same section.
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Bartholini, G., Stadler, H., Lloyd, K.G.: Cholinergic-dopaminergic relation in different brain structures. In: Frontiers in catecholamine research, edit. by E. Usdin and S. Snyder, pp. 741–745. London: Pergamon Press 1973
Battenberg, E.L.F., Bloom, F.E.: A rapid, simple and more sensitive method for the demonstration of central catecholamine neurons and axons by glyoxylic acid induced fluorescence. I. Specificity. Psychopharmacol. Commun. 1, 3–13 (1975)
Bialowas, J., Narkiewicz, O.: Acetylcholinesterase activity in the septum of the rat: A histochemical topography. Acta Neurobiol. Exp. 34, 573–582 (1974)
Bloom, F.E., Battenberg, E.L.F.: A rapid simple and more sensitive method for the demonstration of central catecholamine-containing neurons and axons by glyoxylic acid induced fluorescence: II. A detailed description of methodology. J. Histochem. Cytochem. 24, 561–571 (1976)
Butcher, L.L., Talbot, K., Bilezikjian, L.: Localization of acetylcholinesterase within dopamine containing neurons in the zona compacta of the substantia nigra. Proc. West. Pharmacol. Soc. 18, 256–259 (1975)
Carlsson, A., Falck, B., Hillarp, N.-Å.: Cellular localization of brain monoamines. Acta. physiol. scand. Suppl. 196, 1–27 (1962)
Ehinger, B., Falck, B.: Noradrenaline and cholinesterase in concomitant nerve fibres in iris. Life Sci. 4, 2097–2100 (1965)
Ehinger, B., Falck, B.: Concomitant adrenergic and parasympathetic fibres in the rat iris. Acta physiol. scand. 67, 201–207 (1966)
Ehinger, B., Sporrong, B., Stenevi, U.: Combining the catecholamine fluorescence and methylene blue staining methods for demonstrating nerve fibres. Life Sci. 6, 1973–1974 (1967)
Ehinger, B., Sporrong, B., Stenevi, U.: Simultaneous demonstration of adrenergic and non-adrenergic nerve fibres. Histochemie 13, 105–110 (1968)
Eränkö, O.: Demonstration of catecholamines and cholinesterases in the same section. Pharmacol. Rev. 18, 353–358 (1966)
Eränkö, O., Räisänen, L.: Fibres containing both noradrenaline and acetylcholinesterase in the nerve net of the rat iris. Acta physiol. scand. 63, 505–506 (1965)
Falck, B.: Observations on the possibilities of the cellular localization of monoamines by a fluorescence method. Acta physiol. scand. 56, Suppl. 197 (1962)
Falck, B., Hillarp, N.-Å., Thieme, G., Torp, A.: Fluorescence of catecholamines and related compounds condensed with formaldehyde. J. Histochem. Cytochem. 10, 348–354 (1962)
Fuxe, K.: Evidence for the existence of monoamine neurons in the central nervous system. IV. The distribution of monoamine nerve terminals in the central nervous system. Acta physiol. scand. 64, Suppl. 247 (1965)
Harkmark, W., Mellgren, S.I., Srebro, B.: Acetylcholinesterase histochemistry of the septal region in rat and human: Distribution of enzyme activity. Brain Res. 95, 281–289 (1975)
Holmstedt, B.: A modification of the thiocholine method for the determination of cholinesterase. Acta physiol. scand. 40, 331–357 (1957)
Hökfelt, T., Ljungdahl, Å.: Modification of the Falck-Hillarp formaldehyde fluorescence method using the vibratome®: Simple, rapid and sensitive localization of catecholamines in sections of unfixed or formalin fixed brain tissue. Histochemie 29, 325–339 (1972)
Hökfelt, T., Ljungdahl, Å., Johansson, O., Lindblom, D.: The Vibratome. A useful tool in transmitter histochemistry. In: Amine fluorescence histochemistry, edit. by M. Fujiwara and C. Tanaka, pp. 2–12. Tokyo: Igaku Shoin Ltd 1974
Jacobowitz, D., Koelle, G.B.: Histochemical correlations of acetylcholinesterase and catecholamines in postganglionic autonomic nerves of the cat, rabbit, and guinea pig. J. Pharmacol. exp. Ther. 148, 225–237 (1965)
Jacobowitz, D.M., Palkovits, M.: Topographic atlas of catecholamine and acetylcholinesterase-containing neurons in the rat brain. I. Forebrain (telencephalon, diencephalon). J. comp. Neurol. 157, 13–28 (1974)
Koelle, G.B.: The histochemical localization of cholinesterases in the central nervous system of the rat. J. comp. Neurol. 100, 211–228 (1954)
Koelle, G.B.: Cytological distributions and physiological functions of cholinesterases. In: Cholinesterases and anticholinesterase agents, Heffter-Heubner Handbook Exp. Pharm., Suppl. 15, pp. 187–298, Berlin-Göttingen-Heidelberg: Springer 1963
Koelle, G.B., Friedenwald J.S.: A histochemical method for localizing cholinesterase activity. Proc. Soc. exp. Biol. (N.Y.) 70, 617–622 (1949)
Lewis, P.R., Shute, C.C.D.: The cholinergic limbic system: projections to hippocampal formation, medial cortex, nuclei of the ascending cholinergic reticular system, and the subfornical organ and supra-optic crest. Brain 90, 521–542 (1967)
Lindvall, O.: Mesencephalic dopaminergic afferents to the lateral septal nucleus of the rat. Brain Res. 87, 89–95 (1975)
Lindvall, O., Björklund, A.: The glyoxylic acid fluorescence histochemical method: a detailed account of the methodology for the visualization of central catecholamine neurons. Histochemistry 39, 97–127 (1974)
Palkovits, M., Jacobowitz, D.M.: Topographic atlas of catecholamine and acetylcholinesterase-containing neurons in the rat brain. II. Hindbrain (mesencephalon, rhombencephalon). J. comp. Neurol. 157, 29–42 (1974)
Shute, C.C.D., Lewis, P.R.: The ascending cholinergic reticular system: neocortical, olfactory and subcortical projections. Brain 90, 497–520 (1967)
Torre, J.C. de la, Surgeon, J.W.: A methodological approach to rapid and sensitive monoamine histofluorescence using a modified glyoxylic acid technique: the SPG method. Histochemistry, in press (1976)
Watson, S.J., Barchas, J.D.: Histofluorescence in the unperfused CNS by cryostat and glyoxylic acid: a preliminary report. Psychopharmacol. Commun. 1, 523–531 (1975)
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Lindvall, O. Combined visualization of central catecholamine-and acetylcholinesterase-containing neurons: Application of the glyoxylic acid and thiocholine histochemical methods to the same Vibratome section. Histochemistry 50, 191–196 (1977). https://doi.org/10.1007/BF00491066
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DOI: https://doi.org/10.1007/BF00491066