Summary
Growth cones of ‘differentiating’ neuroblastoma cells in monolayer culture were studied by time-lapse cinematography and electron microscopy. Morphological differentiation, and thus growth cone formation, was induced by the glucocorticoid dexamethasone. Growth cones lengthened gradually at an average rate of 30 μm/h, advancing in stages that involved alternating extensions and retractions of the filopodia and lamellar sheets. During neurite growth the cell body usually remained stationary. The ultrastructure of growth cones was typified by several filopodia, each containing a bundle of microfilaments, agranular endoplasmic reticulum, aggregates of large agranular vesicles lying adjacent to filopodia (previously termed vesicle-filled mounds), many dense-cored vesicles, 100–140 nm in diameter, microtubules, bizarre and distorted mitochondria, and scattered free ribosomes. Comparing the findings with previous ultrastructural accounts of growth cones of cultured ganglion cells, similarities outnumbered differences. The organization of the microfilament bundles and the abundance of free ribosomes were remarkable in the neuroblastoma cell as was the profusion of dense-cored vesicles which were most numerous in the proximal portion of the growth cone.
Similar content being viewed by others
References
Augusti-Tocco, G., Sato, G. H., Claude, P. &Potter, D. (1970) Clonal cell lines of neurons. InControl Mechanisms in the Expression of Cellular Phenotypes (edited byPadykula, H. A.), pp. 109–120. New York: Academic Press.
Birks, R., Mackey, M. C. &Weldon, P. R. (1972) Organelle formation from pinocytotic elements in neurites of cultured sympathetic ganglion.Journal of Neurocytology 1, 311–40.
Booher, J., Sensenbrenner, M. &Mandel, P. (1973) Neuroblastoma cell differentiation: A tissue culture study using time-lapse cinematography.Neurobiology 3, 335–8.
Bray, D. (1970) Surface movements during the growth of single explanted neurons.Proceedings of the National Academy of Sciences (U.S.A.) 65, 905–10.
Bray, D. (1973) Model for membrane movements in the neural growth cone.Nature 244, 93–6.
Breakefield, X. O., Neale, E. A., Neale, J. H. &Jacobwitz, D. M. (1975) Localized catecholamine storage associated with granules in murine neuroblastoma cells.Brain Research 92, 237–56.
Bunge, M. B. (1973) Fine structure of nerve fibers and growth cones of isolated sympathetic neurons in culture.Journal of Cell Biology 56, 713–35.
Bunge, M. B. (1977) Initial endocytosis of peroxidase or ferritin by growth cones of cultured nerve cells.Journal of Neurocytology 6, 407–39.
Chang, C.-M. &Goldman, R. D. (1973) The localization of actin-like fibers in cultured neuroblastoma cells as revealed by heavy meromyosin binding.Journal of Cell Biology 57, 867–74.
Goldman, R. D. (1975) The use of heavy meromyosin binding as an ultrastructural cytochemical method for localizing and determining the possible functions of actin-like microfilaments in nonmuscle cells.Journal of Histochemistry and Cytochemistry 23, 529–42.
Haffke, S. C. &Seeds, N. W. (1975) Neuroblastoma: theE. coli of neurobiology?Life Sciences 16, 1649–57.
Hinkley, R. E. &Telser, A. G. (1974) The effects of halothane on cultured mouse neuroblastoma cells. I. Inhibition of morphological differentiation.Journal of Cell Biology 63, 531–40.
Letourneau, P. C. &Wessells, N. K. (1974) Migratory cell locomotionversus nerve axon elongation. Differences based on the effects of lanthanum ion.Journal of Cell Biology 61, 56–69.
Lodin, Z., Booher, J. &Kasten, F. H. (1970) Phase-contrast cinematographic study of dissociated neurons from embryonic chick dorsal root ganglia cultured in the Rose chamber.Experimental Cell Research 60, 27–39.
Marchisio, P. C., Osborn, M. &Weber, K. (1978) The intracellular organization of actin and tubulin in cultured C-1300 mouse neuroblastoma (clone NB41A3).Journal of Neurocytology 7, 571–82.
Pomerat, C. M., Hendelman, W. J., Raiborn, C. W., JR &Massey, J. F. (1967) Dynamic activities of nervous tissuein vitro. InThe Neuron (edited byHyden, H.), pp. 119–178. New York: Elsevier.
Prasad, K. N. &Hsie, A. W. (1971) Morphological differentiation of mouse neuroblastoma cells inducedin vitro by dibutyryl adenosine 3′: 5′-cyclic monophosphate.Nature New Biology 233, 141–2.
Prasad, K. N., Manual, B., Waymire, J. C., Lees, G. J., Vernadakis, A. &Weiner, N. (1973) Basal level of neurotransmitter synthesizing enzymes and effect of cyclic AMP agents on the morphological differentiation of isolated neuroblastoma clones.Nature New Biology 241, 117–9.
Rees, R. P. &Reese, T. S. (1979) Absence of mounds in freeze-substituted neuritic growth cones.Anatomical Record 193, 661 (abstract).
Roisen, F., Inczedy-Marcsek, M., Hsu, L. &Yorke, W. (1978) Myosin: immunofluorescent localization in neuronal and glial cultures.Science 199, 1445–8.
Ross, J., Olmsted, J. B. &Rosenbaum, J. L. (1975) The ultrastructure of mouse neuroblastoma cells in tissue culture.Tissue and Cell 7, 107–36.
Sandquist, D., Williams, T. H., Sahu, S. K. &Kataoka, S. (1978) Morphological differentiation of a murine neuroblastoma clone in monolayer culture induced by dexamethasone.Experimental Cell Research 113, 375–81.
Schubert, D., Humphreys, S., Baroni, C. &Cohn, M. (1969)In vitro differentiation of a mouse neuroblastoma.Proceedings of the National Academy of Sciences (U.S.A.) 64, 316–23.
Seeds, N. W., Gilman, A. G., Amano, T. &Nirenberg, M. W. (1970) Regulation of axon formation by clonal lines of a neural tumor.Proceedings of the National Academy of Sciences (U.S.A.) 66, 160–7.
Small, J. V., Isenberg, G. &Celts, J. E. (1978) Polarity of actin at the leading edge of cultured cells.Nature 272, 638–9.
Spooner, B. S., Luduena, M. A. &Wessells, N. K. (1974) Membrane fusion in the growth cone-microspike region of embryonic nerve cells undergoing axon elongation in cell culture.Tissue and Cell 6, 399–409.
Tennyson, V. M. (1970) The fine structure of the axon and growth cone of the dorsal root neuroblast of the rabbit embryo.Journal of Cell Biology 44, 62–79.
Weldon, P. R. (1975) Pinocytotic uptake and intracellular distribution of colloidal thorium dioxide by cultured sensory neurites.Journal of Neurocytology 4, 341–56.
Wessells, N. K., Luduena, M. A., Letourneau, P. C., Wrenn, J. T. &Spooner, B. S. (1974) Thorotrast uptake and transit in embryonic glia, heart fibroblasts and neuronsin vitro.Tissue and Cell 6, 757–76.
Yamada, K. M., Spooner, B. S. &Wessells, N. K. (1971) Ultrastructure and function of growth cones and axons of cultured nerve cells.Journal of Cell Biology 49, 614–35.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kataoka, S., Sandquist, D., Williams, L. et al. Growth cones in differentiated neuroblastoma: A time-lapse cinematographic and electron microscopic study. J Neurocytol 9, 591–602 (1980). https://doi.org/10.1007/BF01205027
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01205027