Synopsis
Pineal glands of adult albino rats were examined histochemically using, first, formaldehyde-induced fluorescence to study monoamines and, second, copper thiocholine or copper ferrocyanide methods to study acetylcholinesterase and non-specific cholinesterase by light and electron microscopy. Cholinesterase was determined quantitatively by a constant pH titration assay.
Fluorescent and acetylcholinesterase-positive nerve nets formed identical patterns. Nonspecific cholinesterase was observed only in nerve trunks outside the pineal. Bilateral removal of superior cervical ganglia resulted in complete disappearance of fluorescence and acetylcholinesterase from nerve fibres. Electron microscopically, acetylcholinesterase was found on sympathetic axons containing small granular vesicles. Quantitative cholinesterase determinations suggested that the pineal activity was mainly due to acetylcholinesterase. Comparison of the incubation times required for equal histochemical acetylcholinesterase reactions suggested that the activity of the sympathetic nerve fibres in the pineal is of the same order of magnitude as that in the nerve fibres of the iris.
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References
Arstila, A. (1966). Electron microscopic studies on the structure and histochemistry of the pineal gland of the rat.Neuroendocrin. Suppl. 2, 1–101.
Arstila, A. &Hopsu, V. K. (1964). Studies on the rat pineal gland. I. Ultrastructure.Annls Acad. Sci. fenn. A. 113, 1–21.
Arvy, L. (1961). Contribution à la connaissance de la glande pinéale de Bos taurus L., d'Ovis aries L. et de Sus scrofa L.C. r. Séance Acad. Sci. (Paris) 253, ii, 1361–3.
Bondareff, W. (1965). Submicroscopic morphology of granular vesicles in sympathetic nerves of rat pineal body.Z. Zellforsch. mikrosk. Anat. 67, 211–18.
Burn, J. H. &Rand, M. J. (1965). Acetylcholine in adrenergic transmission.A. Rev. Pharmac. 5, 163–82.
Eränkö, O. (1967a). Histochemistry of nervous tissues: catecholamines and cholinesterases.A. Rev. Pharmac. 7, 203–22.
Eränkö, O. (1967b). The practical histochemical demonstration of catecholamines by formaldehyde-induced fluorescence.Jl R. microsc. Soc. 87, 259–76.
Eränkö, O. &Härkönen, M. (1964). Noradrenaline and acetylcholinesterase in sympathetic ganglion cells of the rat.Acta physiol. scand. 61, 299–300.
Eränkö, O., Härkönen, M., Kokko, A. &Räisänen, L. (1964). Histochemical and starch-gel electrophoretic characterization of desmo- and lyo-esterases in the sympathetic and spinal ganglia of the rat.J. Histochem. Cytochem. 12, 570–81.
Eränkö, O., Koelle, G. B. &Räisänen, L. (1967). A thiocholine-lead ferrocyanide method for acetylcholinesterase.J. Histochem. Cytochem. 15, 674–9.
Eränkö, O. &Räisänen, L. (1965). Fibres containing both noradrenaline and acetylcholinesterase in the nerve net of the rat iris.Acta physiol. scand. 63, 505–6.
Eränkö, O. &Teräväinen, H. (1967). Cholinesterases and eserine-resistant carboxylic esterases in degenerating and regenerating motor end plates of the rat.J. Neurochem. 14, 947–54.
Gomori, G. (1952). Enzymes. InMicroscopic Histochemistry. Principles and Practise, pp. 137–221. Chicago: University of Chicago Press.
Grillo, M. A. (1966). Electron microscopy of sympathetic tissues.Pharmac. Rev. 18, 387–99.
Hökfelt, T. (1968).In vitro studies on central and peripheral monoamine neurons at the ultrastructural level.Z. Zellforsch mikrosk. Anat. 91, 1–74.
Holmstedt, B., Lundgren, G. &Sjöqvist, F. (1962). Determination of acetylcholinesterase activity in normal and denervated sympathetic ganglia of the cat. A biochemical and histochemical comparison.Acta physiol. scand. 57, 187–298.
Jaim-Etcheverry, G. &Zieher, L. M. (1968). Cytochemistry of 5-hydroxytryptamine at the electron microscope level.Z. Zellforsch mikrosk. Anat. 86, 393–400.
Kappers, J. (1960). The development, topographical relations and innervation of the epiphysis cerebri in the albino rat.Z. Zellforsch. mikrosk. Anat. 52, 163–215.
Kappers, J. (1965). Survey of the innervation of the epiphysis cerebri and the accessory pineal organs of vertebrates.Prog. Brain Res. 10, 87–153.
Karnovsky, M. J. &Roots, L. (1964). A ‘direct-coloring’ thiocholine method for cholinesterases.J. Histochem. Cytochem. 12, 219–21.
Koelle, G. B. (1951). The elimination of enzymatic diffusion artifacts in the histochemical localization of cholinesterases and a survey of their cellular distribution.J. Pharmac. exp. Ther. 103, 153–71.
Kokko, A., Mautner, H. G. &Barrnett, R. J. (1969). Fine structural localization of acetylcholinesterase using acetyl-β-methylthiocholine and acetylselenocholine as substrates.J. Histochem. Cytochem. 17, 625–40.
La Bella, F. S. &Shin, S. (1968). Estimation of cholinesterase and choline acetyltransferase in bovine anterior pituitary, posterior pituitary, and pineal body.J. Neurochem. 15, 335–42.
Machado, A. B. M. & Lemos, V. P. J. (1970). Histochemical evidence for a cholinergic sympathetic innervation in the rat pineal body.J. Neuro-visceral Relations (in press).
Owman, C. (1964). New aspects of the mammalian pineal gland.Acta physiol. scand. 63, Suppl. 240, 1–40.
Pellegrino de Iraldi, A., Zieher, L. M. & de Robertis, B. (1965). Ultrastructure and pharmacological studies of nerve endings in the pineal organ; in J. Ariëns Kappers & J. P. Schade's ‘Structure and Function of the Epiphysis Cerebri’.Prog. Brain Res. 10, 389–422.
Rechardt, L. (1969). Electron microscopic and histochemical observations on the supraoptic nucleus of normal and dehydrated rats.Acta physiol. scand. 77 Suppl. 329, 1–79.
Robinson, P. M. (1969). A cholinergic component in the innervation of the longitudinal smooth muscle of the guinea pig vas deferens.J. Cell Biol. 41, 462–76.
Thiéblot, L., Bastide, P., Blaise, S., Boyer, J. &Dastugue, G. (1966). De l'équipement enzymatique de la glande pinéale. Étude critique et expérimentale.Ann. Endocrl 27, 1–20.
Wartman, S. A., Branch, B. J., George, R. &Newman Taylor, A. (1969). Evidence for a cholinergic influence on pineal hydroxyindole o-methyl-transferase activity with changes in environmental lighting.Life Sci. 8, 123–70.
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Eränkö, O., Rechardt, L., Eränkö, L. et al. Light and electron microscopic histochemical observations on cholinesterase-containing sympathetic nerve fibres in the pineal body of the rat. Histochem J 2, 479–489 (1970). https://doi.org/10.1007/BF01003126
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DOI: https://doi.org/10.1007/BF01003126