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A quantitative analysis of ultrastructural changes induced by electrical stimulation of identified spinal cord axons in the larval lamprey

  • Published:
Journal of Neurocytology

Summary

Microelectrodes filled with horseradish peroxidase (HRP) were used to label single identified giant axons in the isolated lamprey spinal cord. Subsequent to the iontophoretic injection of HRP, the spinal cord was stimulated at repetition rates of 20–30/s and the activity in labelled axons monitored. Immediately following failure of the action potential, the spinal cord was fixed by immersion and processed for light and electron microscopy. Electron micrographs were taken of synaptic contacts made by the labelled axons. Several quantitative measures were made from each synapse using a digitizing tablet interfaced with a digital computer. These measures included vesicle number (VN), vesicle area (VA), length of differentiated membrane (DM), vesicle density (VD=VN/VA), vesicle frequency (VF = VN/DM), and a relative measure of the amount of vesicle membrane added to the axolemma during the stimulation period, the curvature ratio (CR). Measures from 106 stimulated synapses were compared with 134 synapses from injected but unstimulated giant axons. The results from these experiments suggest that measurable ultrastructural changes occur during transmitter release at identified C.N.S. synapses, which are consistent with the hypothesis of synaptic vesicle recycling.

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References

  • Akert, K., Moor, H., Pfenninger, K. &Sandri, C. (1969) Contributions of new impregnation methods and freeze etching to the problem of synaptic fine structure.Progress in Brain Research 31, 223–40.

    Google Scholar 

  • Akert, K., Pfenninger, K., Sandri, C. &Moor, H. (1972) Freeze etching and cytochemistry of vesicles and membrane complexes in synapses of the central nervous system. InStructure and function of Synapses (edited byPappas, G. D. andPurpura, D. P.), pp. 67–86. Raven Press: New York.

    Google Scholar 

  • Akert, K. &Sandri, C. (1970) Identification of the active synaptic region by means of histochemical and freeze etching techniques. InExcitatory Synaptic Mechanisms (edited byAndersen, P. andJansen, J. K. S.), pp. 27–41. Universitetsforlaget: Oslo.

    Google Scholar 

  • Atwood, H. L., Lang, F. &Morin, W. A. (1972) Synaptic vesicles: selective depletion in crayfish excitatory and inhibitory axons.Science 176, 1353–5.

    Google Scholar 

  • Bennett, M. V. L., Model, P. G. &Highstein, S. M. (1975) Stimulation induced depletion of vesicles, fatigue of transmission, and recovery processes at a vertebrate central synapse. InCold Spring Harbor Symposia on Quantitative Biology 40, 25–35.

    Google Scholar 

  • Birks, R. I. (1971) Effects of stimulation on synaptic vesicles in sympathetic ganglia, as shown by fixation in the presence of magnesium.Journal of Physiology 216, 26P.

    Google Scholar 

  • Birks, R. I. (1974) The relationship of transmitter release and storage to fine structure in a sympathetic ganglion.Journal of Neurocytology 3, 133–60.

    Google Scholar 

  • Ceccarelli, B. &Hurlbut, W. P. (1975) The effects of prolonged repetitive stimulation in hemicholinium on the frog neuromuscular junction.Journal of Physiology 247, 163–88.

    Google Scholar 

  • Ceccarelli, B., Hurlbut, W. P. &Mauro, A. (1972) Depletion of vesicles from frog neuromuscular junctions by prolonged tetanic stimulation.Journal of Cell Biology 54, 30–58.

    Google Scholar 

  • Ceccarelli, B., Hurlbut, W. P. &Mauro, A. (1973) Turnover of transmitter and synaptic vesicles at the frog neuromuscular junction.Journal of Cell Biology 57, 499–524.

    Google Scholar 

  • Christensen, B. N. (1974) Procion brown: an intracellular dye for light and electron microscopy.Science 1255–6.

  • Christensen, B. N. (1976) Morphological correlates of synaptic transmission in lamprey spinal cord.Journal of Neurophysiology 39, 197–212.

    Google Scholar 

  • Christensen, B. N. &Ebner, F. E. (1978) The synaptic architecture of neurons in opossum somatic sensory-motor cortex: a combined anatomical and physiological study.Journal of Neurocytology 7, 39–60.

    Google Scholar 

  • Christensen, B. N. &Quinn, J. (1979) The relationship between some measures of synaptic ultrastructure as a function of distance from the soma on lamprey reticulospinal neurons.Journal of Neurocytology 8, 737–750.

    Google Scholar 

  • Christensen, B. N. &Teubl, W. P. (1979) Localization of synaptic input on dendrites of a lamprey spinal cord neurone from physiological measurements of membrane properties.Journal of Physiology 297 (in press).

  • Clark, A. W., Hurlbut, W. P. &Mauro, A. (1972) Changes in the fine structure of the neuromuscular junction of the frog caused by black widow spider venom.Journal of Cell Biology 52, 1–14.

    Google Scholar 

  • Del Castillo, J. &Katz, B. (1954) Quantal components of the endplate potential.Journal of Physiology 124, 560–73.

    Google Scholar 

  • Del Castillo, J. &Katz, B. (1956) Biochemical aspects of neuro-muscular transmission.Progress in Biophysics and Biophysical Chemistry 6, 121–70.

    Google Scholar 

  • Fatt, P. &Katz, B. (1950) Some observations on biological noise.Nature 166, 597–8.

    Google Scholar 

  • Fatt, P. &Katz, B. (1952) Spontaneous subthreshold activity at motor nerve endings.Journal of Physiology 117, 109–28.

    Google Scholar 

  • Gennaro, J. F., Nastuk, W. L. &Rutherford, D. J. (1978) Reversible depletion of synaptic vesicles induced by application of high external potassium to the frog neuromuscular junction.Journal of Physiology 280, 237–47.

    Google Scholar 

  • Gray, E. G. (1959) Axosomatic and axodendritic synapses of the cerebral cortex: an electron microscopic study.Journal of Anatomy 193, 420–33.

    Google Scholar 

  • Gray, E. G. (1963) Electron microscopy of presynaptic organelles of the spinal cord.Journal of Anatomy 97, 101–6.

    Google Scholar 

  • Gray, E. G. &Willis, R. A. (1970) On synaptic vesicles, complex vesicles and dense projections.Brain Research 24, 149–68.

    Google Scholar 

  • Heuser, J. E. &Reese, T. S. (1973) Evidence for recycling of synaptic vesicle membrane during transmitter release at the frog neuromuscular junction.Journal of Cell Biology 57, 315–44.

    Google Scholar 

  • Heuser J. E., Reese, T. S. &Landis, D. M. D. (1974) Functional changes in frog neuromuscular junctions studied with freeze fracture.Journal of Neurocytology 3, 109–31.

    Google Scholar 

  • Holtzman, E., Freeman, A. R. &Kashner, A. (1971) Stimulation-dependent alteration in peroxidase uptake at lobster neuromuscular junctions.Science 173, 733–6.

    Google Scholar 

  • Jones, S. F. &Kwanbunbumpen, S. (1970) The effects of nerve stimulation and hemicholinium on synaptic vesicles at the mammalian neuromuscular junction.Journal of Physiology 207, 31–50.

    Google Scholar 

  • Kaneseki, T. &Kadota, K. (1969) The ‘Vesicle in a basket.’Journal of Cell Biology 42, 202–20.

    Google Scholar 

  • Korneliussen, H. (1972) Ultrastructure of normal and stimulated motor endplates.Zeitschrift für Zellforschung und mikroskopische Anatomie 130, 28–57.

    Google Scholar 

  • Lavail, J. H. &Lavail, M. M. (1972) Retrograde axonal transport in the central nervous system.Science 176, 1416–8.

    Google Scholar 

  • Model, P. G., Highstein, S. M. &Bennett, M. V. L. (1975) Depletion of vesicles and fatigue of transmission at a vertebrate central synapse.Brain Research 98, 209–28.

    Google Scholar 

  • Parducz, A., Fehér, O. &Joó, F. (1971) Effects of stimulation and hemicholinium on the fine structure of nerve endings in the superior cervical ganglia of the cat.Brain Research 34, 61–72.

    Google Scholar 

  • Pfenninger, K. (1971a)Freeze-etching of presynaptic membranes: ‘Synaptopores’ as a morphological basis of transmitter release? Inaugural Dissertation, University of Zurich.

  • Pfenninger, K. (1971b) The cytochemistry of synaptic densities. II. Proteinaceous components and mechanisms of synaptic connectivity.Journal of Ultrastructure Research 35, 451–75.

    Google Scholar 

  • Pfenninger, K. (1973) Synaptic morphology and cytochemistry.Progress in Histochemistry and Cytochemistry 5, 1–86.

    Google Scholar 

  • Pfenninger, K., Akert, K., Moor, H. &Sandri, C. (1971) Freeze-fracturing of presynaptic membranes in the central nervous system.Philosophical Transactions of the Royal Society of London Series B 261, 387.

    Google Scholar 

  • Pfenninger, K., Akert, K., Moor, H. &Sandri, C. (1972) The fine structure of freeze-fractured presynaptic membranes.Journal of Neurocytology 1, 129–49.

    Google Scholar 

  • Pfenninger, K. H. &Rovainen, C. M. (1974) Stimulus and calcium dependence of vesicle attachment sites in the presynaptic membrane.Brain Research 72, 1–23.

    Google Scholar 

  • Pumplin, D. W. &Reese, T. S. (1977) Action of brown widow spider venom and botulinum toxin on the frog neuromuscular junction examined with the freeze-fracture technique.Journal of Physiology,273, 443–57.

    Google Scholar 

  • Pysh, J. J. &Wiley, R. G. (1972) Morphological alterations of synapses in electrically stimulated superior cervical ganglia of the cat.Science 176, 191–3.

    Google Scholar 

  • Pysh, J. J. &Wiley, R. G. (1974) Synaptic vesicle depletion and recovery in cat sympathetic ganglia electrically stimulatedin vivo.Journal of Cell Biology 60, 365–74.

    Google Scholar 

  • Rovainen, C. M. (1967) Physiological and anatomical studies on large neurons of central nervous system of the sea lamprey (Petromyzon marinus) II. Dorsal cells and giant interneurons.Journal of Neurophysiology 30, 1024–42.

    Google Scholar 

  • Snow, P. J., Rose, P. K. &Brown, A. G. (1976) Tracing axons and axon collaterals of spinal neurons using intracellular injection of horseradish peroxidase.Science 191, 312–3.

    Google Scholar 

  • Westrum, L. E. (1965) On the origin of synaptic vesicles in cerebral cortex.Journal of Physiology 179, 4–6P.

    Google Scholar 

  • Whittaker, V. P. (1970) The vesicle hypothesis. InExcitatory Synaptic Mechanisms (edited byAndersen, P. andJansen, J. K. S.), pp. 67–76. Universitetsforlaget: Oslo.

    Google Scholar 

  • Wickelgren, W. O. (1975) Structural presynaptic changes following repetitive action potential activity in giant reticulospinal axons of lamprey.Society for Neuroscience Abstracts.

  • Wickelgren, W. O. (1977) Physiological and anatomical characteristics of reticulospinal neurones in lamprey.Journal of Physiology 270, 89–114.

    Google Scholar 

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Author notes

  1. Paul Kershaw

    Present address: School of Medicine, Tufts University, Boston, Massachusetts, USA

Authors and Affiliations

  1. Department of Physiology and Biophysics, University of Texas Medical Branch, 77550, Galveston, Texas, USA

    Paul Kershaw & Burgess N. Christensen

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  1. Paul Kershaw
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  2. Burgess N. Christensen
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Kershaw, P., Christensen, B.N. A quantitative analysis of ultrastructural changes induced by electrical stimulation of identified spinal cord axons in the larval lamprey. J Neurocytol 9, 119–138 (1980). https://doi.org/10.1007/BF01205231

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  • DOI: https://doi.org/10.1007/BF01205231

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