The preparation and characterization of synaptic vesicles of high purity

Brain Res. 1976 Jun 11;109(2):285-309. doi: 10.1016/0006-8993(76)90531-x.


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

  • Acetylcholine / analysis
  • Adenosine Triphosphatases / metabolism
  • Adenosine Triphosphate / analysis
  • Animals
  • Cell Fractionation / methods
  • Centrifugation, Zonal
  • Cerebral Cortex / ultrastructure*
  • Chromatography
  • Electric Organ / ultrastructure*
  • Fishes
  • Guinea Pigs
  • Lipids / analysis
  • Synaptic Vesicles* / analysis
  • Synaptic Vesicles* / enzymology


  • Lipids
  • Adenosine Triphosphate
  • Adenosine Triphosphatases
  • Acetylcholine