Summary
Studies were performed on spherules of approximately 100–300 μm in diameter obtained from in vitro cultures of reaggregates of embryonic fragments of cerebellum from 10–12 day-incubated chick embryos, dissociated with trypsin and cultivated in a rotating shaker for a maximum of 21 days. The differentiated neurons within these spherules included a few Purkinje cells, many granule cells and type II Golgi cells, as well as many glial cells. Zones rich in synaptic knobs and other simple synaptic structures as well as complex synaptic systems with numerous active points of contact, were visible in various parts of the spherule. Typical “glomeruli” consisting of a varicosity or “rosette” joined to the dendritic claws of the granule cells, and “en marron” systems with perikarya of type II Golgi cells were easily recognised. The complete absence of extracerebellar afferents confirms that both the granule and Golgi cells are capable of making synaptic connections with afferents different from those normally formed by extracerebellar mossy or climbing fibres.
The experimental findings confirm that the recipient neurons determine the specific synaptic pattern regardless of the nature of the afferents, and furthermore demonstrate that the clinging activity of the recipient neuron determines the synaptogenic behaviour of nervous pathways.
Similar content being viewed by others
References
Bodian, D.: A model of synaptic and behavioural ontogeny. In: Neuroscience, pp. 129–140. New York: Rockefeller Univ. Press 1970
Cataldi, E., Ieradi, L.A., Caravita, S.: Acetylcholinesterase distribution in the cerebellar cortex in the chick during development. An electron microscopy study. J. Neurocytol. 3, 35–48 (1974)
Chan-Palay, V., Palay, S.L.: Tendril and glomerular collaterals of climbing fibers in the granular layer of the rat's cerebellar cortex. Z. Anat. Entwickl.-Gesch. 133, 247–273 (1971)
Coghill, G.E.: Anatomy and the problem of behaviour. Cambridge: Cambridge Univ. Press 1929
Del Cerro, M.M., Snider, R.S.: Axosomatic and axodendritic synapses in the cerebellum of the newborn rat. Brain Res. 43, 581–586 (1972)
Eccles, J.C., Ito, M., Szentágothai, J.: The cerebellum as a neuronal machine. Berlin-Heidelberg-New York: Springer 1967
Forssman, J.: Zur Kenntnis des Neurotropismus. Beitr. path. Anat. 27, 407–430 (1900)
Gray, E.G.: Axo-somatic and axo-dendritic synapses of the cerebellar cortex. J. Anat. (Lond.) 93, 420–433 (1959)
Gray, E.G.: The granule cells, mossy synapses and Purkinje spine synapses of the cerebellum: light and electron microscope observations. J. Anat. (Lond.) 95, 345–346 (1961)
Hámori, J., Szentágothai, J.: Participation of Golgi neuron processes in the cerebellar glomeruli. An electron microscope study. Exp. Brain Res. 2, 35–48 (1966)
Hámori, J., Szentágothai, J.: Identification of synapses formed in the cerebellar cortex by Purkinje axon collaterals: An electron microscope study. Exp. Brain Res. 5, 118–128 (1968)
Hanna, R.B., Hirano, A., Pappas, G.D.: Membrane specializations of dendritic spines and glia in the weaver mouse cerebellum: a freeze-fracture study. J. Cell Biol. 68, 403–410 (1976)
Kappers Ariens, C.U.: Weitere Mitteilungen über Neurobiotaxis. Folia Neurob. 1 (1908)
Kappers Ariens, C.U.: Dixième contribution à la théorie de la neurobiotaxis. Encéphale 1, 1 (1922)
Karnovsky, M.J.: A formaldehyde-glutaraldehyde fixative of high osmolality for use in electron microscopy. J. Cell Biol. 27, 137A (1965)
Kim, K.U.: Observations on cerebellar granule cells in tissue culture. Z. Zellforsch. 107, 454–465 (1970)
Kim, K.U.: Electron microscope study of mouse cerebellum in tissue culture. Exp. Neurol. 33, 30–44 (1971)
Kim, K.U.: Formation of unattached spines of Purkinje cell dendrites in organotypic cultures of mouse cerebellum. Brain Res. 88, 52–58 (1975)
Ieradi, L.A., Cataldi, E.: Localizzazione della colinesterasi nel cervelletto di uccelli. Riv. Istoch. 10, 413–420 (1964)
Ieradi, L.A., Cataldi, E.: Attività dell'acetilcolinesterasi nel cerveletto di polio coltivato in vitro. Rend. Accad. Naz. Lincei 53, 717–719 (1973)
Jansen, J., Brodal, A.: Aspects of cerebellar anatomy. Oslo: J. Grunt, Tanum Forlay (1955)
Larramendi, L.M.H.: Synaptogenic period of mossy terminals in mouse cerebellum. An electron microscopic study. Anat. Rec. 157, 275 (1967)
Larramendi, L.M.H.: Analysis of synaptogenesis in the cerebellum of the mouse. In: Neurobiology of cerebellar evolution and development (R. Llinas, ed.). Chicago: Am. Med. Ass. Ed. res. Found 1969
Mugnaini, G.: Ultrastructural aspects of cerebellar morphology in the chick embryo. Anat Rec. 154, 391 (1966)
Mugnaini, G.: Cytology and histology of the cerebellar cortex. In: O. Larsell: The comparative anatomy and histology of the cerebellum, Vol. III (Jansen, ed). Minneapolis: Univ. of Minnesota Press 1969
Mugnaini, G.: Ultrastructural studies on the cerebellar histogenesis. II. Maturation of nerve cell populations and establishment of synaptic connections in the cerebellar cortex of the chick. R. Llinas, ed.: Neurobiology of cerebellar evolution and development. Chicago 1969
Mugnaini, G.: Neurones as synaptic targets. In: Excitatory synaptic mechanism (P. Andersen and J.K. Jansen, eds.). Oslo: Universitietfolaget 1970
Mugnaini, G.: Developmental aspects of synaptology with special emphasis upon the cerebellar cortex. In: Cellular aspects of neural growth and differentiation. D.C. Pease, ed.: UCLA Forum in Medical Science, Vol. 14. Los Angeles 1971
Mugnaini, G., Forstrønen, P.F.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)
Palay, S.L.: The electron microscopy of the glomeruli cerebellosi. In: Cytology of nervous tissue. Proc. Anat. Soc. of Brit. and Irel.; Taylor and Francis, ed. London 1961
Palay, S.L., Chan-Palay, V.: Cerebellar cortex. Berlin-Heidelberg-New York: Springer 1974
Privat, A., Drian, M.J.: Specificity of the formation of the mossy fibre — granule cell synapse in the rat cerebellum. An in vitro study. Brain Res. 88, 518–524 (1975)
Privat, A., Drian, M.J.: Postnatal maturation of rat Purkinje cells cultivated in the absence of two afferent systems. An ultrastructure study. J. comp. Neurol. 166, 201–244 (1976)
Privat, A., Drian, M.J., Mandon, P.: The outgrowth of rat cerebellum in organised culture. Z. Zellforsch. 149, 291–308 (1973)
Privat, A., Drian, M.J., Mendon, P.: Synaptogenesis in the outgrowth of rat cerebellum in organised culture. J. comp. Neurol. 153, 291–308 (1974)
Rakic, P.: Neuro-glia relationship during granule cell migration in developing cerebellar cortex. A Golgi and electron microscopic study in Macacus rhesus. J. comp. Neurol. 141, 283–312 (1971)
Rakic, P., Sidmann, R.L.: Organization of cerebellar cortex secondary to deficiency of granule cells in weaver mice. J. comp. Neurol. 152, 133–161 (1973)
Rakic, P., Sidman, R.L.: Sequence of developmental abnormality of weaver mice cerebellum. J. comp. Neurol. 152, 103–134 (1973)
Ramón y Cajal, S.: Neue Darstellung vom histologischen Bau des Zentralnervensystems. Arch. Anat. Physiol. (Anat. Abth.) 319–428 (1893)
Ramón y Cajal, S.: Structure et connections des neurones. Nobel Price Confer. Stockholm (1906)
Ramón y Cajal, S.: Histologie du système nerveux de l'homme et des vertébrés, Vol. I et II. Paris: Maloine 1909–1911
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)
Seil, F.G., Herndon, R.M.: Cerebellar granule cells in vitro. J. Cell Biol. 45, 212–220 (1970)
Shin-Ho Chung: Synaptic remodelling in the mutant cerebellum. Nature (Lond.) 257, 86–87 (1975)
Sotelo, C.: Anatomical, physiological and biochemical studies of the cerebellum from mutant mice. Brain Res. 94, 19–44 (1975)
Sotelo, C., Changeux, J.P.: Bergmann fibres and granular cell migration in the cerebellum of homozygous weaver mutant mouse. Brain Res. 77, 484–491 (1934)
Snider, R.S.: Alterations which occur in mossy terminals of the cerebellum following transection of brachium pontis. J. comp. Neurol. 64, 417–435 (1946)
Stefanelli, A.: I fenomeni della determinazione, della regolazione e del differenziamento del sistema nervoso. Mem. Accad. Naz. Lincei, Ser. VIII, 1, 23–114 (1947)
Stefanelli, A.: Fattore spaziale e organizzazione strutturale nel differenziamento dei neuroni dei lobi ottici e del cervelletto in impianti allanto-coriali. Rend. Accad. Naz. Lincei 16, 792–798 (1954)
Stefanelli, A., Caravita, S.: Ultrastruttura dei sistemi sinaptici del neurone di Mauthner di un teleosteo (Brachydanio rerio). Z. Zellforsch. 62, 1–15 (1964)
Stefanelli, A., Zacchei, A.M., Cataldi, E., Caravita, S., Ieradi, L.A.: Sinapsi in vitro da cellule disgregate di retina di embrione di polio. Rend. Accad. Naz. Lincei 40, 758–762 (1966)
Stefanelli, A., Zacchei, A.M., Caravita, S., Cataldi, E., Ieradi, L.A.: Differenziamento in aggregati di retina in vitro. Arch. Zool. It. 51, 985–995 (1966)
Stefanelli, A., Zacchei, A.M., Caravita, S., Cataldi, E., Ieradi, L.A.: New-forming retinal synapses in vitro. Experientia (Basel) 23, 199 (1967)
Uchizono, K.: Synaptic organization of the Purkinje cells in the cerebellum of the cat Exp. Brain Res. 4, 97–113 (1967)
Uchizono, K.: Inhibitory and excitatory synapses in vertebrate and invertebrate animals. In: Structure and function of inhibitory neural mechanismus (C. von Ruler, S. Skoglund and U. Soderberg, eds.), pp. 33–59. Oxford: Pergamon 1968
Weiss, P.: In vitro experiments on the factors determining the course of the outgrowing nerve fibre. J. exp. Zool. 68, 393–448 (1934)
Weiss, P.: Nerve patterns: the mechanism of nerve growth. Growth (Suppl.) 5, 163–203 (1941)
Wolf, M.K.: Differentiation of neural types and synapses in myelinating cultures of mouse cerebellum. J. Cell Biol. 22, 259–269 (1974)
Wolf, M.K.: Anatomy of cultured mouse cerebellum: II. Organotypic migration of granule cells demonstrated by silver impregnation of normal and mutant cultures. J. comp. Neurol. 140, 281–298 (1970)
Wolf, M.K., Dubois-Dalq, M.: Anatomy of cultured mouse cerebellum: I. Golgi and electron microscopic demonstration of granule cells, their afferent and efferent synapses. J. comp. Neurol. 140, 261–280 (1970)
Author information
Authors and Affiliations
Additional information
This work was supported by a grant from the Consiglio Nazionale delle Ricerche. Technical assistance was given by D. Scorsini for the ultrathin sectioning and electron microscopy, and by G. Gentili for the in vitro preparation and micrographs
Rights and permissions
About this article
Cite this article
Stefanelli, A., Cataldi, E. & Ieradi, L.A. Specific synaptic systems in reaggregated spherules from dissociated chick cerebellum cultivated in vitro. Cell Tissue Res. 182, 311–325 (1977). https://doi.org/10.1007/BF00219767
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00219767