Summary
In long-term organized cultures of newborn mouse cerebellum, granule cell neurons were studied with silver impregnation and electron microscopy.
In silver impregnated cultures, small neurons are defined as granule cell neurons from their size, morphology and location. There are also occasional large nerve endings with the morphology typical of mossy fiber endings.
In correlative electron micrographs, granule cell neurons revealed a faithful reproduction of characteristic structures seen in vivo. The fine structural details of cerebellar glomeruli and myelinated granule cell bodies developed in vitro were also described.
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Allerand, C. D., Murray, M. R.: Myelin formation in vitro. Endogenous influences on cultures of newborn mouse cerebellum. Arch. Neurol. (Chic.) 19, 191–201 (1968).
Bornstein, M. B., Appel, S. H.: The application of tissue culture to the study of experimental allergic encephalomyelitis. 1. Patterns of demyelination. J. Neuropath. exp. Neurol. 20, 141–157 (1962).
—: Tissue culture studies of demyelination. Ann. N.Y. Acad. Sci. 122, 280–286 (1965).
—, Murray, M. R.: Serial observations on patterns of growth, myelin formation, maintenance and degeneration in cultures of newborn rat and kitten cerebellum. J. biophys. biochem. Cytol. 5, 499–504 (1958).
Bunge, M. B., Bunge, R. P., Peterson, E. R.: The onset of synapses formation in spinal cord cultures as studied by electron microscopy. Brain Res. 6, 728–749 (1967).
Bunge, R. P., Bunge, M. B., Peterson, E. R.: An electron microscope study of cultured rat spinal cord. J. Cell Biol. 24, 163–191 (1965).
Callas, G., Hild, W.: Electron microscopic observations of synaptic endings in cultures of mammalian central nervous tissue. Z. Zellforsch. 63, 686–691 (1964).
Crain, S. M.: Development of ‘organotypic’ bioelectric activities in central nervous tissue during maturation in culture. In: Int. Rev. Neurobiol., vol. 9, p. 1–43, ed. by C. C. Pfeiffer and J. R. Smythies. New York: Academic Press 1966.
—, Peterson, E. R.: Complex bioelectric activity in organized tissue cultures of spinal cord (human, rat and chicken). J. Cell comp. Physiol. 64, 1–13 (1964).
Eccles, J. C., Ito, M., Szentágothai, J.: The cerebellum as a neuronal machine. Berlin-Heidelberg-New York: Springer 1967.
Field, E. J., Hughes, D., Raine, C. S.: Observations of the development of myelin in cultures of neonatal rat cerebellum. J. Neurol. Sci. 8, 49–60 (1968).
Fox, C. A., Hillman, D. E., Siegesmund, K. A., Dutta, C. R.: The primate cerebellar cortex: A Golgi and electron microscopic study. In: Progress in brain research, vol. 25, p. 174–225, ed. by C. A. Fox and R. S. Snider. Amsterdam: Elsevier 1967.
Gray, E. G.: The granule cells, mossy synapses and Purkinje spine synapses of the cerebellum: light and electron microscope observations. J. Anat. (Lond.) 95, 345–356 (1961).
Herndon, R. M.: The fine structure of the rat cerebellum. II. The stellate neurons, granule cells, and glia. J. Cell Biol. 23, 277–293 (1964).
Hild, W.: Myelinogenesis in cultures of mammalian central nervous tissue. Z. Zellforsch. 46, 71–95 (1957).
—: Myelin formation around central neurons in vitro. Tex. Rep. Biol. Med. 21, 207–213 (1963).
—: Cell types and neuronal connections in cultures of mammalian central nervous tissue. Z. Zellforsch. 69, 155–188 (1966).
—, Tasaki, I.: Morphological and physiological properties of neurons and glial cells in tissue culture. J. Neurophysiol. 25, 277–304 (1962).
Hoskin, F. C. G., Allerand, C. D.: Metabolism of specifically labelled glucose by explants of newborn mouse cerebellum. J. Neurochem. 15, 427–432 (1968).
Jansen, J., Brodal, A.: Aspects of cerebellar anatomy. Oslo: J. Grundt Tanum Forlag 1954.
Kim, S. U.: Neurons in the tissue culture. Arch. Histol. Jap. 23, 401–429 (1963).
—: Neurons in the tissue culture. Observations on terminal boutons in cultures of mammalian central nervous tissue. Arch. Histol. Jap. 25, 371–381 (1965).
—: Histochemical demonstration of oxidative enzymes associated with carbohydrate metabolism in cerebellar neurons cultured in vitro. Arch. Histol. Jap. 27, 465–471 (1966).
—: Response of neuronal lysosomes to anoxia in tissue culture of mammalian cerebellum. Experientia (Basel) 25, 72–73 (1969).
—: Cytochemical demonstration of ‘marker’ enzymes in nerve cells cultured in vitro. Experientia (Basel) 26, 292–293 (1970).
—, Hirano, A., Murray, M. R.: Neurons with myelinated soma in organized cultures of mouse cerebellum. J. Cell Biol. 43, 68a (1969).
- Murray, M. R., Tourtellotte, W. W., Parker, J. A.: Demonstration in tissue culture of myelinotoxicity in cerebrospinal fluid and brain extracts from multiple sclerosis patients. J. Neuropath, exp. Neurol. (in press).
Lumsden, C. E.: The clinical neuropathology of multiple sclerosis. In: Multiple sclerosis. A reappraisal, p. 243–393, by D. McAlpine, C. E. Lumsden and E. D. Acheson. Baltimore: Williams & Wilkins 1965.
—: Nervous tissue in culture. In: The structure and function of nervous tissue, vol. 1, p. 68–141, ed. by G. H. Bourne. New York: Academic Press 1968.
Mizuno, N., Kim, S. U., Okamoto, M.: Study on embryonal granule cells of the cerebellum. Identification in the tissue culture. Arch. Histol. Jap. 23, 185–211 (1962).
Mugnaini, E., Forstrønen, F.: Ultrastructural studies on the cerebellar histogenesis. I. Differentiation of granule cells and development of glomeruli in the chick embryo. Z. Zellforsch. 77, 115–143 (1967).
Murray, M. R.: Nervous tissues in vitro. In: Cells and tissues in culture. Method, biology and physiology, vol. 2, p. 373–455, ed. by E. N. Willmer. New York: Academic Press 1965.
Peterson, E. R., Crain, S. M., Murray, M. R.: Differentiation and prolonged maintenance of bioelectrically active spinal cord cultures (rat, chick and human). Z. Zellforsch. 66, 130–154 (1965).
Piper, R.: Microtomy techniques and the problem of relation of cell outgrowth to events within explants in vitro. J. Med. Lab. Technol. 19, 1–18 (1962).
Pomerat, C. M., Costero, I.: Tissue culture of cat cerebellum. Amer. J. Anat. 99, 211–247 (1956).
Ramon y Cajal, S.: Histologie du système nerveux de l'homme et des vertébrés Tome 2. Translated by L. Azoulay. Paris: Maloine 1911.
—: Degeneration and regeneration of the nervous system. Translated by R. M. May. New York: Hafner 1959.
Ross, L. L., Bornstein, M. B., Lehrer, G. M.: Electron microscopic observations of rat and mouse cerebellum in tissue culture. J. Cell Biol. 14, 19–30 (1962).
Silberberg, D. H., Schutta, H. S.: The effect of unconjugated bilirubin and related pigments on cultures of rat cerebellum. J. Neuropath. exp. Neurol. 26, 572–585 (1967).
Snider, R. S.: Alterations which occur in mossy terminals of the cerebellum following transsection of brachium pontis. J. comp. Neurol. 64, 417–435 (1936).
Stefanelli, A., Zacchei, A. M., Caravita, S., Cataldi, A., Ieradi, L. A.: New-forming retinal synapses in vitro. Experientia (Basel) 23, 199–200 (1967).
Suyeoka, O.: Small myelinated perikarya in the cerebellar granular layer of mammals including man. Experientia (Basel) 24, 472–473 (1968).
—, Okamoto, M.: Granule cells of the mouse cerebellum cultured in vitro: Their identification, degeneration and perikaryal myelin. Arch. Histol. Jap. 27, 117–130 (1966).
Wolf, M. K.: Differentiation of neuronal types and synapses in myelinating cultures of mouse cerebellum. J. Cell Biol. 22, 259–279 (1964).
Yonezawa, T., Bornstein, M. B., Peterson, E. R., Murray, M. R.: A histochemical study of oxidative enzymes in myelinating cultures of central and peripheral nervous tissue. J. Neuropathol. exp. Neurol. 21, 479–487 (1962).
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This work was initiated while the author was a fellow of National Multiple Sclerosis Society in the laboratory of Dr. Margaret R. Murray at the Department of Surgery, Columbia University, New York, U.S.A. This work was supported by the Medical Research Council of Canada.
The author is greatly indebted to Dr. Asao Hirano, Laboratory Division, Montefiore Medical Center, New York, U.S.A., for his contribution in the preparation of certain electronmicrographs. The author wishes to thank Dr. S. Fedoroff for reading the manuscript.
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Kim, S.U. Observations on cerebellar granule cells in tissue culture. Z. Zellforsch. 107, 454–465 (1970). https://doi.org/10.1007/BF00335434
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DOI: https://doi.org/10.1007/BF00335434