ULTRASTRUCTURE AND FUNCTION OF GROWTH CONES AND AXONS OF CULTURED NERVE CELLS

Dorsal root ganglion nerve cells undergoing axon elongation in vitro have been analyzed ultrastructurally. The growth cone at the axonal tip contains smooth endoplasmic reticulum, vesicles, neurofilaments, occasional microtubules, and a network of 50-A in diameter microfilaments. The filamentous network fills the periphery of the growth cone and is the only structure found in microspikes. Elements of the network are oriented parallel to the axis of microspikes, but exhibit little orientation in the growth cone. Cytochalasin B causes rounding up of growth cones, retraction of microspikes, and cessation of axon elongation. The latter biological effect correlates with an ultrastructural alteration in the filamentous network of growth cones and microspikes. No other organelle appears to be affected by the drug. Removal of cytochalasin allows reinitiation of growth cone-microspike activity, and elongation begins anew. Such recovery will occur in the presence of the protein synthesis inhibitor cycloheximide, and in the absence of exogenous nerve growth factor. The neurofilaments and microtubules of axons are regularly spaced. Fine filaments indistinguishable from those in the growth cone interconnect neurofilaments, vesicles, microtubules, and plasma membrane. This filamentous network could provide the structural basis for the initiation of lateral microspikes and perhaps of collateral axons, besides playing a role in axonal transport.

[1]  S. Carter Effects of Cytochalasins on Mammalian Cells , 1967, Nature.

[2]  B. Spooner,et al.  THE EXPRESSION OF DIFFERENTIATION BY CHICK EMBRYO THYROID IN CELL CULTURE , 1971, The Journal of cell biology.

[3]  M. Abercrombie The bases of the locomotory behaviour of fibroblasts. , 1961, Experimental cell research.

[4]  K. C. Fisher,et al.  Morphological and electrophysiological characteristics of dissociated chick embryonic spinal ganglion cells in culture. , 1969, Experimental neurology.

[5]  G. Kreutzberg Neuronal dynamics and axonal flow. IV. Blockage of intra-axonal enzyme transport by colchicine. , 1969, Proceedings of the National Academy of Sciences of the United States of America.

[6]  D. Bray,et al.  Surface movements during the growth of single explanted neurons. , 1970, Proceedings of the National Academy of Sciences of the United States of America.

[7]  B. Spooner The expression of differentiation by chick embryo thyroid in cell culture. I. Functional and fine structural stability in mass and clonal culture , 1970, Journal of Cellular Physiology.

[8]  S. Cohen,et al.  PURIFICATION OF A NERVE-GROWTH PROMOTING PROTEIN FROM THE MOUSE SALIVARY GLAND AND ITS NEURO-CYTOTOXIC ANTISERUM. , 1960, Proceedings of the National Academy of Sciences of the United States of America.

[9]  A. Hughes The growth of embryonic neurites; a study of cultures of chick neural tissues. , 1953, Journal of anatomy.

[10]  R. Coggeshall,et al.  A SIMPLIFIED LEAD CITRATE STAIN FOR USE IN ELECTRON MICROSCOPY , 1965, The Journal of cell biology.

[11]  R. Snider,et al.  Studies on the developing cerebellums. Ultrastructure of the growth cones , 1968 .

[12]  K. M. Lyser Early differentiation of motor neuroblasts in the chick embryo as studied by electron microscopy. II. Microtubules and neurofilaments. , 1968, Developmental biology.

[13]  J. Metuzals CONFIGURATION OF A FILAMENTOUS NETWORK IN THE AXOPLASM OF THE SQUID (LOLIGO PEALII L.) GIANT NERVE FIBER , 1969, The Journal of cell biology.

[14]  N. K. Wessells,et al.  An ultrastructural study of lens invagination in the mouse. , 1969, The Journal of experimental zoology.

[15]  N. K. Wessells,et al.  Cytochalasin B: effects upon microfilaments involved in morphogenesis of estrogen-induced glands of oviduct. , 1970, Proceedings of the National Academy of Sciences of the United States of America.

[16]  T. Kanaseki,et al.  A Morphological Study of the Coated Vesicle Isolated from the Nerve Endings of the Guinea Pig Brain, with Special Reference to the Mechanism of Membrane Movements , 1969 .

[17]  N. K. Wessells,et al.  MICROFILAMENTS AND CELL LOCOMOTION , 1971, The Journal of cell biology.

[18]  F. O. Schmitt FIBROUS PROTEINS AND NEURONAL DYNAMICS , 1969 .

[19]  H. Wiśniewski,et al.  EFFECTS OF MITOTIC SPINDLE INHIBITORS ON NEUROTUBULES AND NEUROFILAMENTS IN ANTERIOR HORN CELLS , 1968, The Journal of cell biology.

[20]  N. K. Wessells,et al.  Microfilaments in Cellular and Developmental Processes , 1971, Science.

[21]  D. Szőllősi CORTICAL CYTOPLASMIC FILAMENTS OF CLEAVING EGGS: A STRUCTURAL ELEMENT CORRESPONDING TO THE CONTRACTILE RING , 1970, The Journal of cell biology.

[22]  R. G. Harrison The outgrowth of the nerve fiber as a mode of protoplasmic movement. , 1910, The Journal of experimental zoology.

[23]  V. Tennyson THE FINE STRUCTURE OF THE AXON AND GROWTH CONE OF THE DORSAL ROOT NEUROBLAST OF THE RABBIT EMBRYO , 1970, The Journal of cell biology.

[24]  J. Metuzals,et al.  SPATIAL PATTERNS OF THREADLIKE ELEMENTS IN THE AXOPLASM OF THE GIANT NERVE FIBER OF THE SQUID (LOLIGO PEALII L.) AS DISCLOSED BY DIFFERENTIAL INTERFERENCE MICROSCOPY AND BY ELECTRON MICROSCOPY , 1969, The Journal of cell biology.

[25]  A. C. Taylor MICROTUBULES IN THE MICROSPIKES AND CORTICAL CYTOPLASM OF ISOLATED CELLS , 1966, The Journal of cell biology.

[26]  Ross G. Harrison The outgrowth of the nerve fiber as a mode of protoplasmic movement. , 1910 .

[27]  J. Nakai Dissociated dorsal root ganglia in tissue culture. , 1956, The American journal of anatomy.

[28]  P. Weiss,et al.  CHAPTER 21 – NERVE PATTERNS: THE MECHANICS OF NERVE GROWTH1 , 1968 .

[29]  E. Tani,et al.  SUBSTRUCTURE OF MICROTUBULES IN BRAIN NERVE CELLS AS REVEALED BY RUTHENIUM RED , 1970, The Journal of cell biology.

[30]  N. K. Wessells,et al.  Effects of cytochalasin B upon microfilaments involved in morphogenesis of salivary epithelium. , 1970, Proceedings of the National Academy of Sciences of the United States of America.

[31]  J. McIntosh,et al.  INTERMICROTUBULE BRIDGES IN MITOTIC SPINDLE APPARATUS , 1970, The Journal of cell biology.

[32]  E. Nathaniel,et al.  The ultrastructural features of the synapses in the posterior horn of the spinal cord in the rat. , 1966, Journal of ultrastructure research.

[33]  A. Dahlström SYNTHESIS, TRANSPORT, AND LIFE-SPAN OF AMINE STORAGE GRANULES IN SYMPATHETIC ADRENERGIC NEURONS , 1969 .

[34]  K. Porter,et al.  YOLK PROTEIN UPTAKE IN THE OOCYTE OF THE MOSQUITO AEDES AEGYPTI. L , 1964, The Journal of cell biology.

[35]  J. E. Vaughn,et al.  MICROTUBULES AND FILAMENTS IN THE AXONS AND ASTROCYTES OF EARLY POSTNATAL RAT OPTIC NERVES , 1967, The Journal of cell biology.

[36]  K. M. Lyser EARLY DIFFERENTIATION OF MOTOR NEUROBLASTS IN THE CHICK EMBRYO AS STUDIED BY ELECTRON MICROSCOPY. I. GENERAL ASPECTS. , 1964, Developmental biology.

[37]  R. Friede,et al.  Accumulation of axoplasmic organelles in swollen nerve fibers. , 1970, Brain research.