title: The preparation and characterization of synaptic vesicles of high purity.
abstract: Very pure preparations of synaptic vesicles have been obtained from guinea pig cerebral cortex and from the electromotor synapses of Torpedo marmorata by density gradient centrifugation in a zonal rotor followed by chromatography on columns of glass beads of controlled pore size. Markers for soluble cytoplasm (lactate dehydrogenase), plasma and endoplasmic membranes membranes (Na-K-ATPase; acetylcholinesterase, NADPH-cytochrome c reductase], mitochondrial membranes [cytochrome oxidase] and lysosomes [acid phosphatase] were used to assess contamination and were undetectable. The only enzymes detected in the highly purified preparations from guinea pig cerebral cortex were Mg- and Ca-activated ATPases, but their content relative to acetylcholine fell  on chromatography suggesting that they may be constituents of non-cholinergic vesicles. Lipids analyses of the highly purified vesicles confirmed earlier results and showed that glycolipids and lysolecithin are present in negligible amounts; this suggests that lysolecithin is not required for exocytosis of synaptic vesicles. A discussion of the probable limiting concentration of acetycholine in cerebral cortical vesicles derived solely from cholinergic terminals suggests that from 13 to 56% of the vesicles isolated are cholinergic, depending on the assumptions made.
mesh_terms: D000109:Acetylcholine / Q000032:analysis; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000032:analysis; D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D002501:Centrifugation, Zonal; D002540:Cerebral Cortex / Q000648:ultrastructure*; D002845:Chromatography; D004557:Electric Organ / Q000648:ultrastructure*; D005399:Fishes; D006168:Guinea Pigs; D008055:Lipids / Q000032:analysis; D013572:Synaptic Vesicles / Q000032:analysis / Q000201:enzymology
chemical_list: D008055:Lipids; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D000109:Acetylcholine