Summary
Synaptogenesis has been studied in the electric organ of embryonic Torpedo marmorata by use of two antisera directed against components of synaptic vesicles (anti-SV) and presynaptic plasma membranes (ap-anti-TSM), respectively. The anti-SV serum was previously shown to recognize a proteoglycan specific for synaptic vesicles. The ap-anti-TSM serum was raised to plasma membranes of synaptosomes derived from the electromotor nerve terminals and affinity-purified on electric-organ gangliosides. The vesicular antigen was first detectable at the 81-mm stage of development, which is 1–2 weeks earlier than the formation of morphologically mature presynaptic terminals, but is coincident with a rise in choline acetyltransferase levels and the ability of the electric organ to generate discharges. The gangliosidic antigen recognized by the ap-anti-TSM was first detectable on the ventral electrocyte surface at the 93-mm stage of development. This indicates that specific carbohydrate epitopes, not present on the growth cones, are expressed during maturation of the nerve terminal. The nerve terminal components recognized by these sera arose pari passu with neurite coverage of the ventral surface of the electrocyte, reaching a maximum in the adult. In contrast, postsynaptic aggregates of acetylcholine receptor, rendered visible with rhodamine-labeled α-bungarotoxin, arose previous to the presynaptic antigens, reaching a maximum surface density at 110 mm and then declining in the adult.
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Berg DK, Kelly RB, Sargent PB, Williamson P, Hall ZW (1972) Binding of α-bungarotoxin to acetylcholine receptors in mammalian muscle. Proc Natl Acad Sci USA 69:147–151
Blackburn CC, Swank-Hill P and Schnaar RL (1986) Gangliosides support neural retina cell adhesion. J Biol Chem 261:2873–2881
Borroni E, Ferretti P, Fiedler W, Fox GQ (1985) The localization and rate of disappearance of a synaptic vesicle antigen following denervation. Cell Tissue Res 241:367–372
Brockaus M, Magnani JL, Blaszczyk, Steplewski Z, Koprowski H, Karlsson K, Larson G, Ginsburgh V (1981) Monoclonal antibodies directed against the human Leb blood group antigen. J Biol Chem 256:13223–13225
Cartaud J, Benedetti EL, Sobel A, Changeux JP (1978) A morphological study of the receptor protein from Torpedo marmorata in its membrane environment and in its detergent-extracted purified form. J Cell Sci 29:313–337
Ferretti P, Borroni E (1984) The effect of denervation on a cholinergic-specific ganglioside antigen (Chol-1) present in Torpedo electromotor presynaptic plasma membranes. J Neurochem 42:1085–1093
Ferreti P, Borroni E (1986) Putative cholinergic-specific gangliosides in guinea-pig forebrain. J Neurochem 46:1888–1894
Fiedler W, Walker JH, Stadler H, Fox GQ, Whittaker VP (1985) Mechanism of acetylcholine release: exocytosis of synaptic vesicles demonstrated by immunohistochemistry with a vesicle-specific antibody. Biol Chem Hoppe-Seyler 366:133
Fox GQ, Kötting D (1984) Torpedo electromotor system development: a quantitative analysis of synaptogenesis. J Comp Neurol 224:337–343
Fox GQ, Richardson GP (1979) The developmental morphology of Torpedo marmorata: electric organ — electrogenic phase. J Comp Neurol 185:293–315
Grunwald GB, Fredman P, Magnani JL, Trisler D, Ginsburg V, Nirenberg M (1985) Monoclonal antibody 18 B8 detects gangliosides associated with neuronal differentiation and synapse formation. Proc Natl Acad Sci USA 82:4008–4012
Heuser JE, Salpeter SR (1979) Organization of acetylcholine receptors in quick-frozen, deep-etched and rotary-replicated Torpedo postsynaptic membrane. J Cell Biol 82:150–173
Irwin LN (1984) Phylogeny and ontogeny of vertebrate brain gangliosides. In: Ledeen RW, Yu RK, Rapport MM, Suzuki K (eds) Advances in Experimental Medicine and Biology, Plenum Press New York, Vol 174, pp 319–329
Krenz WD, Tashiro T, Wächtler K, Whittaker VP, Witzemann V (1980) Aspects of the chemical embryology of the electromotor system of Torpedo marmorata with special reference to synaptogenesis. Neuroscience 5:617–624
Ledeen RW (1983) Gangliosides. In: Lajtha A (ed) Handbook of Neurochemistry, Plenum Press, New York, London, Vol 3, pp 41–90
Levine JM, Beasley L, Stallcup WB (1984) The D 1.1 antigen: a cell surface marker for germinal cells of the central nervous system J Neurosci 4:820–831
Marchase RB (1977) Biochemical investigation of retinotectal adhesive specificity. J Cell Biol 75:237–257
Max SR, Nelson PG, Brady RO (1970) The effect of denervation on the composition of muscle gangliosides. J Neurochem 17:1517–1520
Ravdin P, Axelrod D (1977) Fluorescent tetramethyl rhodamine derivates of α-bungarotoxin: preparation, separation and characterization. Anal Biochem 80:585–592
Richardson GP, Krenz WD, Kirk C, Fox GQ (1981) Organotypic culture of embryonic electromotor system tissues from Torpedo marmorata. Neuroscience 6:1181–1200
Richardson PJ, Walker JH, Jones RT, Whittaker VP (1982) Identification of a cholinergic-specific antigen Chol-1 as a ganglioside. J Neurochem 38:1605–1614
Sealock R, Wray BE, Froehner SC (1984) Ultrastructural localization of the Mr 43000 protein and the acetylcholine receptor in Torpedo postsynaptic membranes using monoclonal antibodies. J Cell Biol 98:2239–2244
Seybold V, Rahmann H (1985) Changes in developmental profiles of brain gangliosides during ontogeny of a teleost fish (Saroth-erodon mossambicus, Cichlidae). Roux's Arch Dev Biol 194:166–172
Stadler H, Dowe GHC (1982) Identification of a heparan sulphate-containing proteoglycan as a specific core component of cholinergic synaptic vesicles from Torpedo marmorata. EMBO J 1:1381–1384
Svennerholm L (1980) Ganglioside designation. Adv Exp Med Biol 125:11
Tettamanti G, Bonali F, Marchesini S, Zambotti V (1973) A new procedure for the extraction, purification and fractionation of brain gangliosides. Biochim Biophys Acta 296:160–170
Walker JH, Jones RT, Obrocki J, Richardson GP, Stadler H (1982) Presynaptic plasma membranes and synaptic vesicles of cholinergic nerve endings demonstrated by means of specific sera. Cell Tissue Res 223:101–116
Walker JH, Obrocki J, Zimmermann CW (1983) Identification of a proteoglycan antigen characteristic of cholinergic synaptic vesicles. J Neurochem 41:209–216
Willinger M, Schachner M (1980) GMI ganglioside as a marker for neuronal differentiation in mouse. Dev Biol 74:101–107
Witzemann V, Richardson GP, Boustead C (1983) Characterization and distribution of acetylcholine receptors and acetylcholinesterase during electric organ development in Torpedo marmorata. Neuroscience 8:333–349
Young WW, Laine RA, Hakomori SI (1978) Covalent attachment of glycolipids to solid supports and macromlecules. In: Ginsburg V (ed), Methods in Enzymology. Academic Press, New York, Vol 50, pp 137–140
Yu RK (1972) Isolation of gangliosides from electric organ of Electrophorus electricus. J Neurochem 19:2467–2469
Zimmermann H (1979) Vesicle recycling and transmitter release. Neuroscience 4:1773–1804
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Fiedler, W., Borroni, E. & Ferretti, P. An immunohistochemical study of synaptogenesis in the electric organ of Torpedo marmorata by use of antisera to vesicular and presynaptic plasma membrane components. Cell Tissue Res. 246, 439–446 (1986). https://doi.org/10.1007/BF00215906
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DOI: https://doi.org/10.1007/BF00215906