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Ganglioside composition of subcellular fractions,including pre- and postsynaptic membranes,fromTorpedo electric organ
Authors:Robert W Ledeen  Marie-Françoise Diebler  Gusheng Wu  Zi-Hua Lu  Hélène Varoqui
Institution:(1) Departments of Neurosciences and Physiology, UMDNJ New Jersey Medical School, 185 South Orange Avenue, 07103 Newark, New Jersey;(2) Laboratoire de Neurobiologie Cellulaire, Department de Neurochimie, CNRS, 91198 Gif sur Yvette, France
Abstract:Gangliosides were isolated from four subcellular fractions of the electric organ ofTorpedo marmorata: synaptosomes, presynaptic membranes, postsynaptic membranes, and synaptic vesicle membranes. This exploited a principal advantage offered by this tissue: facile separation of pre-and postyynaptic elements. Total ganglioside concentration in presynaptic membranes was approximately twice that of synaptosomes and 15 times that of postsynaptic membranes (47.7, 24.4, and 3.21 mgrg of lipid sialic acid per mg protein, respectively). Synaptic vesicle membranes had the highest overall concentration (78.9) relative to protein, but a concentration approximately comparable to that of presynaptic membranes when expressed relative to phospholipid. The thin-layer patterns of these two fractions were similar, both in terms of total pattern and the specific pattern of gangliotetraose structures as revealed by overlay with cholera toxin B subunit; these were notable for the paucity of monosialo structures and the virtual absence of GM1. Postsynaptic membranes, on the other hand, had a significantly higher content of monosialogangliosides including the presence of GM1. The synaptosomal pattern resembled that of the presynaptic membranes and synaptic vesicles. Thus, a clear difference in ganglioside pattern could be discerned between the pre- and postsynaptic elements of the electric organ.Abbreviations SVs synaptic vesicles - TLC thin-layer chromatography - cholera B-HRP B subunit of cholera toxin linked to horseradish peroxidase
Keywords:Gangliosides  Torpedo marmorata  presynaptic membranes  postsynaptic membranes  synaptic vesicles
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