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Lobster neuromuscular junctions treated with black widow spider venom: Correlation between ultrastructure and physiology

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Journal of Neurocytology

Summary

Black widow spider venom (BWSV) causes marked physiological and morphological alterations at the lobster neuromuscular junction. BWSV is also active at vertebrate neuromuscular junctions but the component which acts on the lobster preparation is different from the one which affects vertebrates. Following exposure to BWSV, lobster neuromuscular junctions showed elevated frequencies of spontaneous miniature synaptic potentials for 15–30 min. Nerve-evoked synaptic potentials became blocked during this period. Subsequently, spontaneous miniature potentials disappeared and less frequent ‘giant’ spontaneous potentials appeared. Ultrastructural examination of excitatory and inhibitory nerve terminals showed that both types were affected by venom treatment. In untreated terminals, synaptic vesicles were grouped near the dense specialized membranes of the synapses. Soon after venom treatment, the synaptic vesicles were dispersed throughout the terminals and many larger and elongated vesicular structures were apparent. At the time of appearance of lsgiant’ spontaneous potentials, few synaptic vesicles were seen in the terminals, but large irregular vacuoles were present. Many mitochondria within the nerve terminals were swollen or disrupted, while nearby muscle mitochondria remained normal in size and appearance. Very few presynaptic dense bodies (‘active zones’) were seen at synapses of affected terminals. The observations are consistent with the hypothesis that BWSV allows an abnormal amount of Ca2+ to enter the nerve terminals, causing the various physiological and morphological changes.

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References

  • Atwood, H. L. (1976) Organization and synaptic physiology of crustacean neuromuscular systems.Progress in Neurobiology 7, 291–391.

    Google Scholar 

  • Atwood, H. L. &Jahromi, S. S. (1978) Fast-axon synapses of a crab leg muscle.Journal of Neurobiology 9, 1–15.

    Google Scholar 

  • Atwood, H. L. &Johnston, H. S. (1968) Neuromuscular synapses of a crab motor axon.Journal of Experimental Zoology 167, 457–70.

    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 

  • Banks, P., Sharpard, M. &Peace, C. (1978) The formation of actin-containing gels by extracts of bovine splenic nerve.Neurosdence 3, 1109–16.

    Google Scholar 

  • Blitz, A. L. &Fine, R. E. (1974) Muscle-like contractile proteins and tubulin in synaptosomes.Proceedings of the National Academy of Sciences (U.S.A.) 71, 4472–6.

    Google Scholar 

  • Blomberg, F., Cohen, R. S. &Siekevitz, P. (1977) The structure of postsynaptic densities isolated from dog cerebral cortex. II. Characterization and arrangement of some of the major proteins within the structure.Journal of Cell Biology 74, 204–25.

    Google Scholar 

  • Ceccarelli, B., Grohovaz, F. &Hurlbut, W. P. (1979) Freeze-fracture studies of frog neuromuscular junctions during intense release of neurotransmitter. I. Effects of black widow spider venom and Ca2+-free solutions on the structure of the active zone.Journal of Cell Biology 81, 163–77.

    Google Scholar 

  • Ceccarelli, B. &Hurlbut, W. P. (1975) Transmitter release and the vesicle hypothesis. InGolgi Centennial Symposium (edited bySantini, M.), pp. 529–545. New York: Raven Press.

    Google Scholar 

  • 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 

  • Clark, A. W., Mauro, A., Longenecker, H. E. &Hurlbut, W. P. (1970) Effects of black widow spider venom on the frog neuromuscular junction. Effects on the fine structure.Nature 225, 703–5.

    Google Scholar 

  • Couteaux, R. &PÉcot-Dechavassine, M. (1970) Vésicules synaptiques et poches au niveau des ‘zones actives’ de la jonction neuromusculaire.Comptes rendus hebdomadaire des séances de l'Academie des sciences, Serie D 271, 2346–9.

    Google Scholar 

  • Cull-Candy, S. G., Neal, H. &Usherwood, P. N. R. (1973) Action of black widow spider venom on an aminergic synapse.Nature 241, 353–4.

    Google Scholar 

  • Del Castillo, J. &Pumplin, D. W. (1975) Discrete and discontinuous action of brown widow spider venom on the presynaptic nerve terminals of frog muscle.Journal of Physiology 252, 491–508.

    Google Scholar 

  • Douglas, W. W. (1974) Involvement of calcium in exocytosis and the exocytosis-vesiculation sequence.Biochemical Society Symposium 39, 1–28.

    Google Scholar 

  • Dreyer, F., Peper, K., Akert, K., Sandri, C. &Moor, H. (1973) Ultrastructure of the ‘active zone’ in the frog neuromuscular junction.Brain Research 62, 373–80.

    Google Scholar 

  • Dudel, J. &Kuffler, S. W. (1961) The quantal nature of transmission and spontaneous miniature potentials at the crayfish neuromuscular junction.Journal of Physiology 155, 514–29.

    Google Scholar 

  • Fritz, L. C., Tzeng, M.-C. &Mauro, A. (1980) Different components of black widow spider venom mediate transmitter release at vertebrate and lobster neuromuscular junctions.Nature 283, 486–7.

    Google Scholar 

  • Fritz, L. C., Wang, C. C. &Gorio, A. (1979) Avermectin B1a irreversibly blocks postsynaptic potentials at the lobster neuromuscular junction by reducing muscle membrane resistance.Proceedings of the National Academy of Sciences (U.S.A.) 76, 2062–6.

    Google Scholar 

  • Frontali, N. (1972) Catecholamine-depleting effect of black widow spider venom on iris nerve fibers.Brain Research 37, 146–8.

    Google Scholar 

  • Frontali, N., Ceccarelli, B., Gorio, A., Mauro, A., Siekevitz, P., Tzeng, M.-C. &Hurlbut, W. P. (1976) Purification from black widow spider venom of a protein factor causing the depletion of synaptic vesicles at neuromuscular junctions.Journal of Cell Biology 68, 461–79.

    Google Scholar 

  • Gorio, A., Hurlbut, W. P. &Ceccarelli, B. (1978) Acetylcholine compartments in mouse diaphragm: Comparison of the effects of black widow spider venom, electrical stimulation and high concentrations of potassium.Journal of Cell Biology 78, 716–33.

    Google Scholar 

  • Gray, E. G. (1975) Presynaptic microtubules and their association with synaptic vesicles.Proceedings of the Royal Society of London, Series B 190, 369–72.

    Google Scholar 

  • Gray, E. G. (1978) Synaptic vesicles and microtubules in frog motor endplates.Proceedings of the Royal Society of London, Series B 203, 219–27.

    Google Scholar 

  • Heuser, J. E., Katz, B. &Miledi, R. (1971) Structural and functional changes of frog neuromuscular junctions in high calcium solutions.Proceedings of the Royal Society of London, Series B 178, 407–15.

    Google Scholar 

  • Heuser, J. E. &Miledi, R. (1971) Effect of lanthanum ions on function and structure of frog neuromuscular junctions.Proceedings of the Royal Society of London, Series B 179, 247–60.

    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. (1977) The origin and fate of secretory packages, especially synaptic vesicles.Neuroscience 2, 327–55.

    Google Scholar 

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

    Google Scholar 

  • Holtzman, E., Teichberg, S., Abrahams, S. J., Citkowitz, E., Crain, S. M., Kawai, N. &Peterson, E. R. (1973) Notes on synaptic vesicles and related structures, endoplasmic reticulum, lysosomes and peroxisomes in nervous tissue and the adrenal medulla.Journal of Histochemistry and Cytochemistry 21, 349–85.

    Google Scholar 

  • Hunter, D. R., Haworth, R. A. &Southard, J. H. (1976) Relationship between configuration, function, and permeability in calcium-treated mitochondria.Journal of Biological Chemistry 251, 5069–77.

    Google Scholar 

  • Ichikawa, A. (1965) Fine structural changes in response to hormonal stimulation of the perfused canine pancreas.Journal of Cell. Biology 24, 369–85.

    Google Scholar 

  • Jahromi, S. S. &Atwood, H. L. (1974) Three-dimensional ultrastructure of the crayfish neuromuscular apparatus.Journal of Cell Biology 63, 599–613.

    Google Scholar 

  • Kawai, N., Mauro, A. &Grundfest, H. (1972) Effect of black widow spider venom on the lobster neuromuscular junctions.Journal of General Physiology 60, 650–64.

    Google Scholar 

  • Longenecker, H. E., Hurlbut, W. P., Mauro, A. &Clark, A. W., (1970) Effects of black widow spider venom on the frog neuromuscular junction. Effects on end-plate potential, miniature end-plate potential and nerve terminal spike.Nature 225, 701–3.

    Google Scholar 

  • Marcum, J. M., Dedmen, J. R., Brinkley, B. R. &Means, A. R. (1978) Control of microtubule assembly-disassembly by calcium-dependent regulator protein.Proceedings of the National Academy of Sciences (U.S.A.) 75, 3771–5.

    Google Scholar 

  • Marotta, C. A., Strocchi, P. &Gilbert, J. M. (1978) Microheterogeneity of brain cytoplasmic synaptoplasmic actins.Journal of Neurochemistry 30, 1441–51.

    Google Scholar 

  • Misler, S. &Hurlbut, W. P. (1979) Action of black widow spider venom on quantized release of acetylcholine at the frog neuromuscular junction: Dependence upon external Mg2+.Proceedings of the National Academy of Sciences (U.S.A.) 76, 991–5.

    Google Scholar 

  • Mushynski, W. E., Glen, S. &Therien, H.-M. (1978) Actin-like and tubulin-like proteins in synaptic junctional complexes.Canadian Journal of Biochemistry 56, 820–30.

    Google Scholar 

  • Olmstead, J. B. &Borisy, G. G. (1975) Ionic and nucleotide requirements for microtubule polymerizationin vitro.Biochemistry 14, 2996–3005.

    Google Scholar 

  • Ornberg, R. L., Smyth, T. &Benton, A. w. (1976) Isolation of a neurotoxin with a presynaptic action from the venom of the black widow spider (Lactrodectus mactans Fabr.)Toxicon 14, 329–33.

    Google Scholar 

  • Papahadjopoulos, D., Portis, A. &Pangborn, W. (1978) Calcium-induced lipid phase transitions and membrane fusion.Annals of the New York Academy of Sciences 308, 50–63.

    Google Scholar 

  • Peachey, L. D. (1964) Electron microscopic observations on the accumulation of divalent cations in intramitochondrial granules.Journal of Cell Biology 20, 95–111.

    Google Scholar 

  • Pécot-Dechavassine, M. &Couteaux, R. (1975) Modifications structurales des terminaisons motrices de muscles de grenouille soumis à l'actions de la vinblastine.Comptes rendus hebdomadaire des séances de l'Académie des sciences, Serie D 280, 1099–101.

    Google Scholar 

  • Pollard, T. D. (1976) The role of actin in the temperature-dependent gelation and contraction of extracts ofAcanthamoeba.Journal of Cell Biology 68, 579–601.

    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 

  • Rees, D. &Usherwood, P. N. R. (1972) Fine structure of normal and degenerating motor axons and nerve-muscle synapses in the locust,Schistocerca gregaria.Comparative Biochemistry and Physiology 43A, 83–101.

    Google Scholar 

  • Rubin, L. L., Gorio, A. &Mauro, A. (1978) Effect of concanavalin A on black widow spider venom activity at the neuromuscular junction: implications for mechanisms of venom action.Brain Research 143, 107–24.

    Google Scholar 

  • Sharoni, Y., Eimerl, S. &Schramm, M. (1976) Secretion of old versus new exportable protein in rat parotid slices. Control by neurotransmitters.Journal of Cell Biology 71, 107–22.

    Google Scholar 

  • Sherman, R. G. &Atwood, H. L. (1971) Structure and neuromuscular physiology of a newly discovered muscle in the walking legs of the lobster,Homarus americanus.Journal of Experimental Zoology 176, 461–74.

    Google Scholar 

  • Smith, J. E., Clark, A. W. &Kuster, T. A. (1977) Suppression by elevated calcium of black widow spider venom activity at frog neuromuscular junctions.Journal of Neurocytology 6, 519–39.

    Google Scholar 

  • Stossel, T. P. &Hartwig, J. H. (1976) Interactions of actin, myosin, and a new actin-binding protein of rabbit pulmonary macrophages. II. Role in cytoplasmic movement and phagocytosis.Journal of Cell Biology 68, 602–19.

    Google Scholar 

  • Suszkiw, J. B. &Whittaker, V. P. (1979) Role of vesicle recycling in vesicular storage and release of acetylcholine inTorpedo electroplaque synapses.Progress in Brain Research 49, 153–62.

    Google Scholar 

  • Tzeng, M.-C. &Siekevitz, P. (1978) The effect of the purified major protein factor (alpha-latrotoxin) of black widow spider venom on the release of acetylcholine and norepinephrine from mouse cerebral cortex slices.Brain Research 139, 190–6.

    Google Scholar 

  • Uchizono, K. (1967) Inhibitory synapses on the stretch receptor neurone of the crayfish.Nature 214, 833–4.

    Google Scholar 

  • Usherwood, P. N. R. (1973) Release of transmitter from degenerating locust motoneurones.Journal of Experimental Biology 59, 1–16.

    Google Scholar 

  • Usherwood, P. N. R. &Rees, D. (1972) Quantitative studies of the spatial distribution of synaptic vesicles within normal and degenerating motor axons of the locust,Schistocerca gregaria.Comparative Biochemistry and Physiology 43A, 103–18.

    Google Scholar 

  • Walters, B. B. &Matus, A. I. (1975) Tubulin in postsynaptic junctional lattice.Nature 257, 496–8.

    Google Scholar 

  • Winlow, W. &Usherwood, P. N. R. (1975) Ultrastructural studies of normal and degenerating mouse neuromuscular junctions.Journal of Neurocytology 4, 377–94.

    Google Scholar 

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Authors and Affiliations

  1. The Rockefeller University, 10021, New York, New York, USA

    L. C. Fritz

  2. Department of Zoology, University of Toronto, M5S 1A1, Toronto, Ontario, Canada

    H. L. Atwood & S. S. Jahromi

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  1. L. C. Fritz
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  2. H. L. Atwood
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  3. S. S. Jahromi
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Fritz, L.C., Atwood, H.L. & Jahromi, S.S. Lobster neuromuscular junctions treated with black widow spider venom: Correlation between ultrastructure and physiology. J Neurocytol 9, 699–721 (1980). https://doi.org/10.1007/BF01205034

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  • DOI: https://doi.org/10.1007/BF01205034

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