Skip to main content
SpringerLink
Log in

Injury-induced remodelling and regeneration of the ribbon presynaptic terminalin vitro

  • Published:
Journal of Neurocytology

Summary

The neuronal response to axonal injury may relate to the type of insult incurred. Recently, neuritic and presynaptic varicosity regeneration by isolated adult salamander photoreceptors was demonstrated. We have used this system to compare the rod photoreceptor response to two types of injury:denervation/detargeting, the removal of pre-and postsynaptic partners from the axon terminal, andaxotomy, the removal of the axon terminal itself. Cells were followed with time-lapse video microscopy for 24–48h in culture and immunolabelled for SV2 or synaptophysin to identify synaptic vesicle-containing varicosities. Although all injured cells responded with regenerative growth, denervated/detargeted photoreceptors (i.e. neurons which retain their axon terminal) grew 80% more processes and fourfold more presynaptic varicosities than axotomized neurons. In cells which retained their original axon and terminal, varicosity formation generally began with axon retraction. Retraction was followed by elaboration of a lamellipodium and, by 48 h, development of varicosity-bearing neurites from the lamellipodium. Synaptic vesicle protein localization in denervated/detargeted cells paralleled axon terminal reorganization. Axotomized cells, in contrast, lacked synaptic vesicle protein immunoreactivity during this period. To detect synaptic protein synthesis, photoreceptors were examined for colocalization of synaptic vesicle protein with rab6, a Golgi marker, by confocal microscopy. As expected, synaptic vesicle protein staining was present in the Golgi complex during regeneration; however, in cells with an axon, new synaptic vesicle protein-labelled varicosities were found at early stages, prior to the appearance of immunolabel in the Golgi complex. The data demonstrate remarkable plasticity in the ribbon synapse, and suggest that in adult rod cells with an intact axon terminal, synaptic vesicle protein synthesis is not a prerequisite for the formation of new presynaptic-like terminals. We propose that preexisting axonal components are reutilized to expedite presynaptic renewal as an early response to denervation/detargeting.

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

  • Aguayo, A. J., Bray, G. M., Rasminsky, M., Zwimpfer, T., Carter, D. &Vidal-Sanz, M. (1990) Synaptic connections made by axons regenerating in the central nervous system of adult mammals.Journal of Experimental Biology 153, 199–224.

    Google Scholar 

  • Anglister, L., Farber, I. C., Shahar, A. &Grinvald, A. (1982) Localization of voltage-sensitive calcium channels along developing neurites: their possible role in regulating neurite elongation.Developmental Biology 94, 351–65.

    Google Scholar 

  • Antony, C., Cibert, C., Géraud, G., Santa Maria, A., Maro, B., Mayau, V. &Goud, B. (1992) The small GTP-binding protein rab6p is distributed from medial Golgi to thetrans Golgi network as determined by a confocal microscopic approach.Journal of Cell Science 103, 785–96.

    Google Scholar 

  • Applegate, M. D., Kerr, D. S. &Landfield, P. W. (1987) Redistribution of synaptic vesicles during long-term potentiation in the hippocampus.Brain Research 401, 401–6.

    Google Scholar 

  • Atwood, H. L., Dixon, D. &Wojtowicz, J. M. (1989) Rapid introduction of long-lasting synaptic changes at crustacean neuromuscular junctions.Journal of Neurobiology 20, 373–85.

    Google Scholar 

  • Bader, C. R., Macleish, P. R. &Schwartz, E. A. (1978) Responses to light of solitary rod photoreceptors isolated from tiger salamander retina.Proceedings of the National Academy of Sciences (USA) 75, 3507–11.

    Google Scholar 

  • Bailey, C. H. &Kandel, E. R. (1993) Structural changes accompanying memory storage.Annual Review of Physiology 55, 397–426.

    Google Scholar 

  • Bailey, C. H., Chen, M., Keller, F. &Kandel, E. R. (1992) Serotonin-mediated endocytosis of apCAM: an early step of learning-related synaptic growth in Aplysia.Science 256, 645–9.

    Google Scholar 

  • Binder, L. I., Frankfurter, A. &Rebhun, L. I. (1985) The distribution of tau in the mammalian central nervous system.Journal of Cell Biology 101, 1371–8.

    Google Scholar 

  • Bray, D. (1970) Surface movements during the growth of single explanted neurons.Proceedings of the National Academy of Sciences (USA) 65, 905–10.

    Google Scholar 

  • Bray, D. (1973) Model for membrane movements in the neural growth cone.Nature 244, 93–6.

    Google Scholar 

  • Brown, M. C., Holland, R. L. &Hopkins, W. G. (1981) Motor nerve sprouting.Annual Review of Neuroscience 4, 17–42.

    Google Scholar 

  • Buckley, K. &Kelly R. B. (1985) Identification of a transmembrane glycoprotein specific for secretory vesicles of neural and endocrine cells.Journal of Cell Biology 100, 1284–94.

    Google Scholar 

  • Chiang, R. G. &Govind, C. K. (1986) Reorganization of synaptic ultrastructure at facilitated lobster neuromuscular terminals.Journal of Neurocytology 15, 63–74.

    Google Scholar 

  • Cooke, I., Graf, R., Grau, S., Haylett, B., Meyers, D. &Ruben, P. (1989) Crustacean peptidergic neurons in culture show immediate outgrowth in simple medium.Proceedings of the National Academy of Sciences (USA) 86, 402–6.

    Google Scholar 

  • Cotman, C. W. &Nadler, J. V. (1978) Reactive synaptogenesis in the hippocampus. InNeuronal plasticity (edited byCotman, C. W.) pp. 227–71. New York: Raven Press.

    Google Scholar 

  • Cotman, C. W. &Nieto-Sampedro, M. (1984) Cell biology of synaptic plasticity.Science 224, 1287–94.

    Google Scholar 

  • Cotman, C. W., Nieto-Sampedro, M. &Harris, E. W. (1981) Synapse replacement in the nervous system of adult vertebrates.Physiological Reviews 61, 684–784.

    Google Scholar 

  • Deretic, D. &Papermaster, D. S. (1993) Rab6 is associated with a compartment that transports rhodopsin from thetrans-Golgi to the site of rod outer segment disk formation in frog retinal photoreceptors.Journal of Cell Science 106, 803–13.

    Google Scholar 

  • Dowling, J. E. (1987)The retina: an approachable part of the brain. Cambridge, MA, London: The Belknap Press of Harvard University Press.

    Google Scholar 

  • Dräger, U. C., Edwards, D. L. &Barnstable, C. J. (1984) Antibodies against filamentous components in discrete cell types of the mouse retina.Journal of Neuroscience 4, 2025–42.

    Google Scholar 

  • Goslin, K. &Banker, G. (1989) Experimental observations on the development of polarity by hippocampal neurons in culture.Journal of Cell Biology 108, 1507–16.

    Google Scholar 

  • Goud, B., Zahraoui, A., Tavitian, A. &Saraste, J. (1990) Small GTP-binding protein associated with Golgi cisternae.Nature 345, 553–6.

    Google Scholar 

  • Gouras, P., Du, J., Gelanze, M., Lopez, R., Kwun, R., Kjeldbye, H. &Krebs, W. (1991a) Survival and synapse formation of transplanted rat rods.Journal of Neural Transplantation and Plasticity 2, 91–100.

    Google Scholar 

  • Gouras, P., Du, J., Kjeldbye, H., Kwun, R., Lopez, R. &Zack, D. J. (1991b) Transplanted photoreceptors identified in dystrophic mouse retina by a transgenic reporter gene.Investigative Ophthalmology and Visual Science 32, 3167–74.

    Google Scholar 

  • Govind, C. K. &Walrond, J. P. (1989) Structural plasticity at crustacean neuromuscular synapses.Journal of Neurobiology 20, 409–21.

    Google Scholar 

  • Greenough, W. T. &Bailey, C. H. (1988) The anatomy of a memory: convergence of results across a diversity of tests.Trends in Neurosciences 11, 142–7.

    Google Scholar 

  • Hicks, D. &Molday, R. S. (1986) Differential immunogold-dextran labeling of bovine and frog rod and cone cells using monoclonal antibodies against bovine rhodopsin.Experimental Eye Research 42, 55–71.

    Google Scholar 

  • Holtzman, E., Augenbraun, E., St. Jules, R. &Santa-hernandez, M. (1993) Endosomes, lysosomes, andtrans-Golgi related systems in conventional neurons and the frog retina.Advances in Cell and Molecular Biology of Membranes 1, 225–67.

    Google Scholar 

  • Jasmin, B. J., Goud, B., Camus, G. &Cartaud, J. (1992) The low molecular weight guanosine triphosphate-binding protein Rab6p associates with distinct post-Golgi vesicles inTorpedo marmorata electrocytes.Neuroscience 49, 849–55.

    Google Scholar 

  • Johnson, G. D., Davidson, R. S., McNamee, K. C., Russel, G., Goodwin, D. &Holbrow, E. J. (1982) Fading of immunofluorescence during microscopy: a study of the phenomenon and its remedy.Journal of Immunological Methods 55, 231–42.

    Google Scholar 

  • Jones, S. M., Crosby, J. R., Salamero, J. &Howell, K. E. (1993) A cytosolic complex of p62 and rab6 associates with TGN38/41 and is involved in budding of exocytic vesicles from thetrans-Golgi network.Journal of Cell Biology 122, 775–88.

    Google Scholar 

  • Lasansky, A. (1973) Organization of the outer synaptic layer in the retina of the larval tiger salamander.Philosophical Transactions of the Royal Society of London. Series B: Biological Sciences 265, 471–89.

    Google Scholar 

  • Ludin, B. &Matus, A. (1993) The neuronal cytoskeleton and its role in axonal and dendritic plasticity.Hippocampus Special Issue 3, 61–72.

    Google Scholar 

  • Macleish, P. R. &Townes-Anderson, E. (1988) Growth and synapse formation among major classes of adult salamander retinal neurons in vitro.Neuron 1, 751–60.

    Google Scholar 

  • Macleish, P. R., Barnstable, C. J. &Townes-Anderson, E. (1983) Use of a monoclonal antibody as a substrate for mature neuronsin vitro.Proceedings of the National Academy of Sciences (USA) 80, 7014–18.

    Google Scholar 

  • Mandell, J. W., Townes-Anderson, E., Czernik, A. J., Cameron, R., Greengard, P. &De Camilli, P. (1990) Synapsins in the vertebrate retina: absence from ribbon synapses and heterogeneous distribution among conventional synapses.Neuron 5, 19–33.

    Google Scholar 

  • Mandell, J. W., Macleish, P. R. &Townes-Anderson, E. (1993) Process outgrowth and synaptic varicosity formation by adult photoreceptorsin vitro.Journal of Neuroscience 13, 3533–48.

    Google Scholar 

  • Martinez, O., Schmidt, A., Salaméro, J., Hoflack, B., Roa, M. &Goud, B. (1994) The small GTP-binding protein rab6 functions in intra-Golgi transport.Journal of Cell Biology 127, 1575–88.

    Google Scholar 

  • Matthews, D. A., Cotman, C. W. &Lynch, G. (1976) An electron microscopic study of lesion-induced synaptogenesis in the dentate gyrus of the adult rat. II. Reappearance of morphologically normal synaptic contacts.Brain Research 115, 1–21.

    Google Scholar 

  • McQuarrie, I. G. &Lasek, R. J. (1989) Transport of cytoskeletal elements from parent axons into regenerating daughter axons.Journal of Neuroscience 9, 436–46.

    Google Scholar 

  • Mosinger Ogilvie, J., Landgraf, M., Lett, J. M., Tenkova, T., Wang, X., Wu, M. &Silverman, M. S. (1994) synaptic connectivity of transplanted photoreceptors in RD mice expressing the transgenic marker LacZ.Investigative Ophthalmology and Visual Science Abstracts 35, 1523.

    Google Scholar 

  • Nachman-Clewner, M. &Townes-Anderson, E. (1994) Influence of the axon on regeneration of presynaptic varicosities by rod photoreceptors.Investigative Ophthalmology and Visual Science Abstracts 35, 1403.

    Google Scholar 

  • Nachman-Clewner, M. &Townes-Anderson, E. (1995) Influence of the axon on presynaptic regeneration by adult sensory neuronsin vitro.Society of Neuroscience Abstracts 21(3), 1769.

    Google Scholar 

  • Palade, G. E. (1975) Intracellular aspects of the process of protein synthesis.Science 189, 347–58.

    Google Scholar 

  • Purves, D. &Lichtman, J. W. (1985)Principles of neural development. Sunderland, MA: Sinauer.

    Google Scholar 

  • Purves, D. &Lichtman, J. W. (1987) Synaptic sites on reinnervated nerve cells visualized at two different times in living mice.Journal of Neuroscience 7, 1492–7.

    Google Scholar 

  • Schmied, R., Huang, C.-C., Zhang, X.-P., Ambron, D. A. &Ambron, R. T. (1993) Endogenous axoplasmic proteins and proteins containing nuclear localization signal sequences use the retrograde axonal transport/nuclear import pathway inAplysia neurons.Journal of Neuroscience 13, 4064–71.

    Google Scholar 

  • Silverman M. S., Hughes, S. E., Valentino, T. L. &Liu, Y. (1992) Photoreceptor transplantation: anatomic, electrophysiologic, and behavioral evidence for the functional reconstruction of retinas lacking photoreceptors.Experimental Neurology 115, 87–94.

    Google Scholar 

  • Sobue, K. (1993) Actin-based cytoskeleton in growth cone activity.Neuroscience Research 18, 91–102.

    Google Scholar 

  • Speidel, C. C. (1941) Adjustments of nerve endings.The Harvey Lectures 36, 126–58.

    Google Scholar 

  • Tashiro, T. &Komiya, Y. (1991) Changes in organization and axonal transport of cytoskeletal proteins during regeneration.Journal of Neurochemistry 56, 1557–63.

    Google Scholar 

  • Tixier-Vidal, A., Faivre-Bauman, A., Picart, R., Loudes, C. &Wiedenmann, B. (1988) Immunoelectron microscopic localization of synaptophysin in a Golgi subcompartment of developing hypothalamic neurons.Neuroscience 26, 847–61.

    Google Scholar 

  • Tixier-Vidal, A., Barret, A., Picart, R., Mayau, V., Vogt, D., Wiedenmann, B. &Goud, B. (1993) The small GTP-binding protein, Rab6p, is associated with both Golgi and post-Golgi synaptophysincontaining membranes during synaptogenesis of hypothalamic neurons in culture.Journal of Cell Science 105, 935–47.

    Google Scholar 

  • Townes-Anderson, E. (1995) Intersegmental fusion in vertebrate rod photoreceptors: rod cell structure revisited.Investigative Ophthalmology and Visual Science 36, 1918–33.

    Google Scholar 

  • Townes-Anderson, E., Macleish, P. R. &Raviola, E. (1985) Rod cells dissociated from mature salamander retina: ultrastructure and uptake of horseradish peroxidase.Journal of Cell Biology 100, 175–88.

    Google Scholar 

  • Townes-Anderson, E., Mandell, J. W. &Macleish, P. R. (1992) Effect of the outer segment on the formation of synaptic varicosities by adult rod cells in vitro.Investigative Ophthalmology and Visual Science Abstracts 33, 1059.

    Google Scholar 

  • Tucker, R. P. &Matus, A. I. (1988) Microtubuleassociated proteins characteristic of embryonic brain are found in the adult mammalian retina.Developmental Biology 130, 423–34.

    Google Scholar 

  • Ullrich, B. &Südhof, T. C. (1994) Distribution of synaptic markers in the retina: implications for synaptic vesicle traffic in ribbon synapses.Journal of Physiology 88, 249–57.

    Google Scholar 

  • Valtorta, F., Jahn, R., Fesce, R., Greengard, P. &Ceccarelli, B. (1988) Synaptophysin (p38) at the frog neuromuscular junction: its incorporation into the axolemma and recycling after intense quantal secretion.Journal of Cell Biology 107, 2717–27.

    Google Scholar 

  • Virtanen, I., Ekblom, P. &Laurila, P. (1980) Subcellular compartmentalization of saccharide moieties in cultured normal and malignant cells.Journal of Cell Biology 85, 429–34.

    Google Scholar 

  • Walters, E. T. &Ambron, R. T. (1995) Long-term alterations induced by injury and by 5-HT inAplysia sensory neurons: convergent pathways and common signals?Trends in Neurosciences 18, 137–42.

    Google Scholar 

  • Wojtowicz, J. M., Marin, L. &Atwood, H. L. (1989) Synaptic restructuring during long-term facilitation at the crayfish neuromuscular junction.Canadian Journal of Physiology and Pharmacology 67, 167–71.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Cornell University Medical College, 10021, New York, NY

    Michele Nachman-Clewner

  2. Department of Neurosciences, University of Medicine and Dentistry of New Jersey, New Jersey Medical School, 07103, Newark, NJ, USA

    Ellen Townes-Anderson

Authors
  1. Michele Nachman-Clewner
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ellen Townes-Anderson
    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

Nachman-Clewner, M., Townes-Anderson, E. Injury-induced remodelling and regeneration of the ribbon presynaptic terminalin vitro . J Neurocytol 25, 597–613 (1996). https://doi.org/10.1007/BF02284827

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02284827

Keywords

Search

Navigation