Skip to main content
SpringerLink
Log in

Early structural changes in the axoplasmic cytoskeleton after axotomy studied by cryofixation

  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Summary

Alterations in the cytoskeleton were studied in the axoplasm of neurites at the tips of proximal stumps of transected chicken sciatic nerves. The studies were carried out using cryofixation with a nitrogen-cooled propane jet. The most immediate effect is the almost complete disassembly of axoplasmic microtubules. This consequently causes the axonal transport of membrane-bounded organelles to cease and results in an accumulation of mitochondria and vesicles of the smooth endoplasmic reticulum. The neurofilament network is partially disorganized. Neurofilaments become shorter and fragmented, and are linked by a large number of anastomosed cross-linkers. The neurofilaments become newly aligned to the axis of the axoplasm and are of normal length 48–72 h after the transsection. At this stage the newly formed neurofilament bundles are in close proximity to the anastomosed cisternae and profiles of the smooth endoplasmic reticulum. The axonal sprouts always show a normally organized cytoskeletal network. These studies support the idea that the rapid remodelling of the neurofilament network is apparently a local event, not dependent on the slow transport of cytoskeletal materials to the tip of the proximal stump. The repair of the degraded cytoskeleton may be in accordance with the function of the endoplasmic reticulum as Ca2+-sequestering membrane system, which may be involved in restoring the physiological conditions of the axoplasm.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Anderson PN, Mitchell J, Mayor D (1978) The role of calcium ions in the synthesis and transport of noradrenalin carrier vesicles in guinea-pig sympathetic neurons in vitro. J Neurocytol 7:623–636

    Google Scholar 

  • Baitinger C, Cheney R, Clements D, Glicksman M, Hirokawa N,.levine J, Meiri K, Simon C, Skene P, Willard M (1983) Axonally transported proteins in axon development, maintenance, and regeneration. Cold Spring Harbor Symp Quant Biol 48 pt 2:791–802

    Google Scholar 

  • Bernhardt R, Huber G, Matus A (1985) Differences in the developmental patterns of three microtubule-associated proteins in the rat cerebellum. J Neurosci 5:977–991

    Google Scholar 

  • Bignami A, Chi NH, Dahl D (1984a) Laminin in rat sciatic nerve undergoing Wallerian degeneration. Immunofluorescence study with laminin and neurofilament antisera. J Neuropathol Exp Neurol 43:94–103

    Google Scholar 

  • Bignami A, Chi NH, Dahl D (1984b) Regenerating dorsal roots and the nerve entry zone: An immunofluorescence study with neurofilament and laminin antisera. Exp Neurol 85:426–436

    Google Scholar 

  • Bisby MA (1978) Fast axonal transport of labeled protein in sensory axons during regeneration. Exp Neurol 61:281–300

    Google Scholar 

  • Blümcke S, Niedorf HR, Rode J (1966) Axoplasmic alterations in the proximal and distal stumps of transsected nerves. Acta Neuropathol 7:44–61

    Google Scholar 

  • Bray D, Bunge MB (1981) Serial analysis of microtubules in cultured rat sensory axons. J Neurocytol 10:589–605

    Google Scholar 

  • Carbonetto S, Muller KJ (1982) Nerve fiber growth and the cellular response to axotomy. Curr Top Dev Biol 17:33–76

    Google Scholar 

  • Carlsen RC (1982) Axonal transport of adenylate cyclase activity in normal and axotomized frog sciatic nerve. Brain Res 232:413–424

    Google Scholar 

  • De Iraldi AP, De Robertis E (1968) The neurotubular system of the axon and the origin of granulated and non-granulated vesicles in regenerating nerves. Z Zellforsch 87:330–344

    Google Scholar 

  • De Vries GH, Salzer JL, Bunge RP (1982) Axolemma-enriched fraction isolated from PNS and CNS are mitogenic for cultured Schwann cells. Dev Brain Res 3:295–299

    Google Scholar 

  • Droz B (1975) Synthetic machinery and axoplasmic transport: Maintenance of neuronal connectivity. In: Tower DB (ed) The nervous system. Vol. 1. Raven Press, New York, pp 111–127

    Google Scholar 

  • Duce IR, Keen P (1976) A light and electron microscope study of changes occurring at the cut ends following section of the dorsal roots of rat spinal nerves. Cell Tissue Res 170:491–505

    Google Scholar 

  • Friede RL, Bischhausen R (1980) The fine structure of stumps of transected nerve fibers in subserial sections. J Neurol Sci 44:181–203

    Google Scholar 

  • Friede RL, Samorajski T (1970) Axon caliber relates to neurofilaments and microtubules in sciatic nerve fibers of rats and mice. Anat Rec 167:379–387

    Google Scholar 

  • Frizell M, Sjöstrand J (1974) The axonal transport of slowly migrating (3H)leucine labelled proteins and the regeneration rate in regenerating hypoglossal and vagus nerves of the rabbit. Brain Res 81:267–283

    Google Scholar 

  • Grafstein B, McQuarrie IG (1978) The role of the nerve cell body in axon regeneration. In: Cotman C (ed) Neuronal Plasticity. Raven Press, New York, pp 155–195

    Google Scholar 

  • Grafstein B, Murray M (1969) Transport of protein in goldfish optic nerve during regeneration. Exp Neurol 25:494–508

    Google Scholar 

  • Hirokawa N (1982) The intramembrane structure of tight junctions: An experimental analysis of the single-fibril and two-fibril models using the quick-freeze method. J Ultrastruct Res 80:288–301

    Google Scholar 

  • Hirokawa N, Glicksman MA, Willard MB (1984) Organization of mammalian neurofilament polypeptides within the neuronal cytoskeleton. J Cell Biol 98:1523–1536

    Google Scholar 

  • Hirokawa N, Bloom GS, Vallee RB (1985) Cytoskeletal architecture and immunocytochemical localization of microtubule-associated proteins in regions of axons associated with rapid axonal transport: The β,β′-iminodipropionitrile-intoxicated axon as a model system. J Cell Biol 101:227–239

    Google Scholar 

  • Hoffman P, Lasek RJ (1975) The slow component of axonal transport. Identification of major structural polypeptides of the axon and their generality among mammalian neurons. J Cell Biol 66:351–366

    Google Scholar 

  • Hoffman PN, Lasek RJ (1980) Axonal transport of the cytoskeleton in regenerating motor neurons: Constancy and change. Brain Res 202:317–333

    Google Scholar 

  • Hoffman PN, Griffin JW, Price DL (1984) Control of axonal caliber by neurofilament transport. J Cell Biol 99:705–714

    Google Scholar 

  • Hoffman PN, Thompson GW, Griffin JW, Price DL (1985) Changes in neurofilament transport coincide temporally with alterations in the caliber of axons in regenerating motor fibers. J Cell Biol 101:1332–1340

    Google Scholar 

  • Kreutzberg GW (1981) Neurobiologische Aspekte der Nervenregeneration. Arch Otorhinolaryngol 231:71–88

    Google Scholar 

  • Kristensson K (1978) Retrograde transport of macromolecules in axons. Ann Rev Pharmacol Toxicol 18:97–110

    Google Scholar 

  • Lasek RJ, Black MM (1977) How do axons stop growing? Some clues from the metabolism of the proteins in the slow component of axonal transport. In: Roberts S (ed) Mechanisms, Regulation and Special Functions of Protein Synthesis in the Brain. Elsevier, Amsterdam, pp 161–169

    Google Scholar 

  • Lasek RJ, Garner JA, Brady ST (1984) Axonal transport of the cytoplasmic matrix. J Cell Biol 99:212s-221s

    Google Scholar 

  • Liem RKH, Yen S-H, Salomon GD, Shelanski ML (1978) Intermediate filaments in nervous tissue. J Cell Biol 79:637–645

    Google Scholar 

  • Malmgren LT, Olsson Y (1979) Early influx of horseradish peroxidase into axons of the hypoglossal nerve during Wallerian degeneration. Neurosci Lett 13:13–18

    Google Scholar 

  • Margaritis LH, Elgsaeter A, Branton D (1977) Rotary replication for freeze-etching. J Cell Biol 72:47–56

    Google Scholar 

  • Matthews MR (1973) An ultrastructural study of axonal changes following constriction of postganglionic branches of the superior cervical ganglion in the rat. Phil Trans R Soc Lond [Biol] 264:479–505

    Google Scholar 

  • Matus A, Bernhardt R, Hugh-Jones T (1981) High molecular weight microtubule-associated proteins are preferentially associated with dendritic microtubules in brain. Proc Natl Acad Sci USA 78:3010–3014

    Google Scholar 

  • Meiri H, Dormann A, Spira ME (1983) Comparison of ultrastructural changes in proximal and distal segments of transected giant fibers of the cockroach Periplaneta americana. Brain Res 263:1–14

    Google Scholar 

  • Meller K (1985) Ultrastructural aspects of cryofixed nerves. Cell Tissue Res 242:289–300

    Google Scholar 

  • Meller K (1987) The cytoskeleton of cryofixed Purkinje cells of the chicken cerebellum. Cell Tissue Res 247:155–165

    Google Scholar 

  • Metuzals J, Montpetit V, Clapin DF (1981) Organization of the neurofilamentous network. Cell Tissue Res 214:455–482

    Google Scholar 

  • Mitchell J, Caren CA (1982) Degeneration of non-myelinated axons in the rat sciatic nerve following lysolecithin injection. Acta Neuropathol 56:187–193

    Google Scholar 

  • Moor H, Kistler J, Müller M (1976) Freezing in a propane jet. Experientia 32:805

    Google Scholar 

  • Morris JH, Hudson AR, Weddell G (1972a) A study of degeneration and regeneration in the divided rat sciatic nerve based on electron microscopy. I. The traumatic degeneration of myelin in the proximal stump of the divided nerve. Z Zellforsch 124:76–102

    Google Scholar 

  • Morris JH, Hudson AR, Weddell G (1972b) A study of degeneration and regeneration in the divided rat sciatic nerve based on electron microscopy. II. The development of the “regenerating unit”. Z Zellforsch 124:103–130

    Google Scholar 

  • Ochs S (1976) Fast axoplasmic transport in the fibres of chromatolysed neurones. J Physiol 255:249–261 (1976)

    Google Scholar 

  • Olsson TP, Forsberg I, Kristensson K (1978) Uptake and retrograde axonal transport of horseradish peroxidase in regenerating facial motor neurons of the mouse. J Neurocytol 7:323–336

    Google Scholar 

  • Payer AF (1979) An ultrastructural study of Schwann cell response to axonal degeneration. J Comp Neurol 183:365–384

    Google Scholar 

  • Perry GW, Wilson DL (1981) Protein synthesis and axonal transport during nerve regeneration. J Neurochem 37:1203–1217

    Google Scholar 

  • Ramon y Cajal S (1928) Degeneration and Regeneration of the Nervous System (translated and edited by RM May). Hafner, New York

    Google Scholar 

  • Romine JS, Aguayo AJ, Bray GM (1975) Absence of Schwann cell migration along regenerating unmyelinated nerves. Brain Res 98:601–606

    Google Scholar 

  • Schlaepfer WW (1974) Calcium-induced degeneration of axoplasm in isolated segments of rat peripheral nerve. Brain Res 69:203–215

    Google Scholar 

  • Schlaepfer WW (1977) Immunological and ultrastructural studies of neurofilaments isolated from rat peripheral nerve. J Cell Biol 74:226–240

    Google Scholar 

  • Schlaepfer WW, Micko S (1978) Chemical and structural changes of neurofilaments in transected rat sciatic nerve. J Cell Biol 78:369–378

    Google Scholar 

  • Schlaepfer WW, Freeman LA (1978) Neurofilament proteins of rat peripheral nerve and spinal cord. J Cell Biol 78:653–662

    Google Scholar 

  • Schliwa M (1986) The Cytoskeleton. An Introductory Survey. Springer, Wien New York

    Google Scholar 

  • Schnapp BJ, Reese TS (1982) Cytoplasmic structure in rapid-frozen axons. J Cell Biol 94:667–679

    Google Scholar 

  • Sotelo C, Taxi J (1973) On the axonal migration of catecholamines in constricted sciatic nerve of the rat. A radioautographic study. Z Zellforsch 138:345–370

    Google Scholar 

  • Theiler RF, McClure WO (1978) Rapid axoplasmic transport of proteins in regenerating sensory nerve fibers. J Neurochem 31:433–447

    Google Scholar 

  • Thomas PK, King RHM (1974) The degeneration of unmyelinated axons following nerve section: an ultrastructural study. J Neurocytol 3:497–512

    Google Scholar 

  • Tohyama K, Ide C, Nitatori T, Yokota R (1983) Nearest-neighbor distance of intermediate filaments in axons and Schwann cells. Distinction between axons and Schwann cell processes in the denervated and reinnervated peripheral nerves. Acta Neuropathol 60:194–198

    Google Scholar 

  • Tsukita S, Usukura J, Tsukita S, Ishikawa H (1982) The cytoskeleton in myelinated axons: A freeze-etch replica study. Neuroscience 7:2135–2147

    Google Scholar 

  • Tytell M, Brady ST, Lasek RJ (1984) Axonal transport of a subclass of tau proteins: Evidence for the regional differentiation of microtubules in neurons. Proc Natl Acad Sci USA 81:1570–1574

    Google Scholar 

  • Venable JH, Coggeshall R (1964) A simplified lead-citrate stain for use in electron microscopy. J Cell Biol 25:407–408

    Google Scholar 

  • Wechsler W, Hager H (1962) Elektronenmikroskopische Untersuchung der Wallerschen Degeneration des peripheren Säugetiernerven. Beitr Pathol Anat Allg Pathol 126:352–380

    Google Scholar 

  • Weiss P, Pillai A (1965) Convection and fate of mitochondria in nerve fibers: Axonal flow as vehicle. Proc Natl Acad Sci USA 54:48–56

    Google Scholar 

  • Willard M, Simon C (1981) Antibody decoration of neurofilaments. J Cell Biol 89:198–205

    Google Scholar 

  • Willard M, Simon C (1983) Modulations of neurofilament axonal transport during the development of rabbit retinal ganglion cells. Cell 35:551–559

    Google Scholar 

  • Willard M, Skene JHP (1982) Molecular events in axonal regeneration. In: Nicholls JG (ed) Repair and Regeneration of the Nervous System, Dahlem Konferenzen 1982. Springer, Berlin Heidelberg New York, pp 71–89

    Google Scholar 

  • Willard M, Simon C, Baitinger C, Levine J, Skene P (1980) Association of an axonally transported polypeptide (H) with 100-A filaments. Use of immunoaffinitiy electron microscope grids. J Cell Biol 85:587–596

    Google Scholar 

  • Zelená J (1971) Neurofilaments and microtubules in sensory neurons after peripheral nerve section. Z Zellforsch 117:191–211

    Google Scholar 

  • Zelená J, Lubinska L, Gutmann E (1968) Accumulation of organelles at the ends of interrupted axons. Z Zellforsch 91:200–219

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut für Anatomie, Abteilung für Cytologie, Ruhr-Universität Bochum, Germany

    Karl Meller

Authors
  1. Karl Meller
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Meller, K. Early structural changes in the axoplasmic cytoskeleton after axotomy studied by cryofixation. Cell Tissue Res. 250, 663–672 (1987). https://doi.org/10.1007/BF00218961

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00218961

Key words

Search

Navigation