Summary
A synaptic vesicle fraction and a synaptic plasma membrane fraction obtained after subfractionation of synaptosomes from chick forebrain have been used to produce antisera in rabbits.
Immunofluorescence histology with the two antisera revealed that they reacted strongly with synaptic terminal regions present in the chick forebrain, cerebellum and spinal cord. In addition, the synaptic plasma membrane antiserum (but not the synaptic vesicle antiserum) reacted with preterminal axons in the cerebellum and spinal cord.
Comparison of the two antisera by two-dimensional immunoelectrophoresis, revealed the presence of common antigens in the synaptosomal vesicle and plasma membrane fractions.
Incubation of synaptosomesin vitro with the synaptosomal vesicle antiserum and complement produced a dose-dependent inhibition of synaptosome swelling up to a maximum of 55% of that obtained with the synaptosomal plasma membrane antiserum. The results of this test are consistent with the hypothesis that some synaptosomal vesicle antigens may be present also in the synaptosomal plasma membrane and imply that they face the external surface of the synaptosomes.
The fate of vesicle membrane components in synaptosomal plasma membranes is not known. The possibility is discussed that they may be recycled locally by a mechanism similar to that proposed by Heuser and Reese (1973) for re-use of synaptic vesicle membranes at the neuromuscular junction.
Similar content being viewed by others
References
Bock, E. andJørgensen, O. S. (1975) Rat brain synaptic vesicles and synaptic plasma membranes compared by crossed immunoelectrophoresis.FEBS Letters 52, 37–9.
Clarke, H. G. M. andFreeman, T. (1968) Quantitative immunoelectrophoresis of human serum proteins.Clinical Science 35, 403–13.
Cotman, C. W. (1974) Isolation of synaptosomal and synaptic plasma membrane fractions. InMethods in Enzymology (edited byFleischer, L. andPacker, L.) 31A pp. 445–52. Academic Press; New York.
Cotman, C. W. andMatthews, D. E. (1971) Synaptic plasma membranes from rat brain synaptosomes. Isolation and partial characterization.Biochimica Biophysica Acta 249, 380–94.
Douglas, W. W. (1973) How do neurones secrete peptides? Exocytosis and its consequences, including ‘synaptic vesicle’ formation, in the hypothalamo-neurohypophyseal system.Progress in Brain Research 39, 21–39.
Fried, R. C. andBlaustein, M. P. (1976) Synaptic vesicle recycling in synaptosomesin vitro.Nature 261, 255–6.
Gurd, J. W., Jones, L. R., Mahler, H. R. andMoore, W. J. (1974) Isolation and partial characterization of rat brain synaptic plasma membranes.Journal of Neuro chemistry 22, 281–90.
Heuser, J. E. andReese, T. S. (1973) Evidence of recycling of synaptic vesicle membrane during transmitter release at the frog neuromuscular junction.Journal of Cell Biology 57, 315–44.
Heuser, J. E., Reese, T. S. andLandis, D. M. D. (1974) Functional changes in frog neuromuscular junctions studied with freeze-fracture.Journal of Neurocytology 3, 109–31.
Howe, P. R. C., Rostas, J. A. P., Fenwick, E. M. andLivett, B. G. (1975) Further evidence for incorporation of synaptic vesicle proteins into synaptosomal plasma membranes.Proceedings of the Australian Bio chemical Society 8, 95.
Hurlbut, W. P. andCeccarelli, B. (1974) Transmitter release and recycling of synaptic vesicle membrane at the neuromuscular junction. InAdvances in Cytopharmacology (edited byCeccarelli, B., Clementi, F. andMeldolesi, J.), 2, pp. 141–54. New York: Raven Press.
Jamieson, J. D. andPalade, G. E. (1971) Condensing vacuole conversion and zymogen granule discharge in pancreatic exocrine cells: metabolicstudies.Journal of Cell Biology 48, 503–22.
Jeffrey, P. L. andGunning, P. (1976) The nature and location of chick synaptosomal plasma membrane RNA. (Submitted for publication.)
Jørgensen, O. S. (1976) Localization of the antigens D1, D2 and D3 in the rat brain synaptic membrane.Journal of Neurochemistry 27, 1223–7.
Jørgensen, O. S. andBock, E. (1974) Brain specific synaptosomal membrane proteins demonstrated by crossed immunoelectrophoresis.Journal of Neurochemistry 23, 879–80.
Livett, B. G., Fenwick, E. M. andEadie, J. (1974a) Ca++-dependent redistribution of antigenic components during membrane fusion.Proceedings of the Australian Biochemical Society 7, 57.
Livett, B. G., Rostas, J. A. P., Jeffrey, P. L. andAustin, L. (1974b) Antigenicity of isolated synaptosomal membranes.Experimental Neurology 43, 330–8.
Livett, B. G., Howe, P. R. C., Fenwick, E. M. andAustin, L. (1975) An immunochemical approach to the transport of axonal vesicular proteins and their release from nerve terminals. InRecent Advances in Myology (edited byBradley, W. G., Gardner-Medwin, D. andWalton, J. N.), pp. 215–23. Amsterdam: Excerpta Medica.
Meldolesi, J. (1974) Membranes and membrane surfaces. Dynamics of cytoplasmic membranes in pancreatic acinar cells.Philosophical Transactions of the Royal Society of London, Series B 268, 39–53.
Morgan, I. G. (1976) Commentary: Synaptosomes and cell separation.Neuroscience 1, 159–65.
Morgan, I. G., Wolfe, L. S., Mandel, P. andGombos, G. (1971) Isolation of plasma membranes from rat brain.Biochimica Biophysica Acta 241 737–51.
Nairn, R. C. (1969)Fluorescent Protein Tracing. London: Livingston.
Pfenninger, K. H. andRovainen, C. M. (1974) Stimulation — and calcium-dependence of vesicle attachment sites in the presynaptic membrane; a freeze-cleave study of the lamprey spinal cord.Brain Research 72, 1–23.
Pfenninger, K., Akert, K., Moor, H. andSandri, C. (1971) Freeze-fracturing of presynaptic membranes in the central nervous system.Philosophical Transactions of the Royal Society of London, Series B 261, 387.
Raiteri, M., Bertollini, A., La Bella, R. (1972) Synaptosome antisera effect permeability of synaptosomal membranesin vitro.Nature New Biology 238, 242–4.
Raiteri, M., Angelini, F., Bertollini, A. andFederico, R. (1973) Quantitative evaluation of antigen antibody interactions at the external surface of the nerve ending membrane.Neurobiology 3, 225–31.
Rostas, J. A. P. andJeffrey, P. L. (1977a) Immunochemical characterization of synaptosomal membrane antigens from chicken brain. I. Histochemical localization in the day-old chick.Neurochemical Research 2, 59–85.
Rostas, J. A. P. andJeffrey, P. L. (1977b) Immunochemical characterization of synaptosomal membrane antigens from chicken brain. II. Histochemical demonstration of changes in distribution during development.Neurochemical Research (In press).
Rostas, J. A. P. andJeffrey, P. L. (1977c) Immunochemical characterization of synaptosomal membrane antigens from chicken brain. III. Quantitative determination and partial characterization.Neurochemical Research (In press).
Tartakoff, A. M., Greene, L. J., Jamieson, J. S. andPalade, G. E. (1974) Parellelism in the processing of pancreatic proteins. InAdvances in Cytopharmacology (edited byCeccarelli, B., Clementi, F. andMeldolesi, J.), 2, pp. 177–94. New York: Raven Press.
Van Leeuwen, C., Stam, H. andOestreicher, A. B. (1976) Isolation and partial characterization of chick brain synaptic plasma membranes.Biochimica Biophysica Acta 436, 53–67.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Howe, P.R.C., Fenwick, E.M., Rostas, J.A.P. et al. Immunochemical comparison of synaptic plasma membrane and synaptic vesicle membrane antigens. J Neurocytol 6, 339–352 (1977). https://doi.org/10.1007/BF01175195
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01175195