The Journal of Cell Biology, Vol 87, 98-103, Copyright © 1980 by The Rockefeller University Press

ARTICLES

An antiserum specific for cholinergic synaptic vesicles from electric organ

SS Carlson and RB Kelly

Rabbit antisera to highly purified synaptic vesicles from the electric organ of Narcine brasiliensis, an electric ray, reveal a unique population of synaptic vesicle antigens in addition to a population shared with other electric organ membranes. Synaptic vesicle antigens were detected by binding successively rabbit antivesicle serum and radioactive goat anti-rabbit serum. To remove antibodies directed against antigens common to synaptic vesicles and other electric organ fractions, the antivesicle serum was extensively preadsorbed against an electric organ membrane fraction that was essentially free of synaptic vesicles. The adsorbed serum retained 40% of its ability to bind to synaptic vesicles, suggesting that about half of the antigenic determinants are unique. Vesicle antigens were quantified with a radioimmunoassay (RIA) that utilized precipitation of antibody-antigen complexes with Staphylococcus aureus cells. By this assay, the vesicles, detected by their acetylcholine (ACh) content and the antigens detected by the RIA, have the same buoyant density after isopycnic centrifugation of crude membrane fractions on sucrose and glycerol density gradients. The ratio of ACh to antigenicity was constant across the vesicle peaks and was close to that observed for vesicles purified to homogeneity. Even though the vesicles make up only approximately 0.5% of the material in the original homogenate, the ratio of acetylcholine to vesicle antigenicity could still be measured and also was indistinguishable from that of pure vesicles. We conclude that synaptic vesicles contain unique antigenic determinants not present to any measurable extent in other fractions of the electric organ. Consequently, it is possible to raise a synaptic vesicle- specific antiserum that allows vesicles to be detected and quantified. These findings are consistent with earlier immunohistochemical observations of specific antibody binding to motor nerve terminals.
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This article has been cited by other articles:

• Ngsee, J.K., Trimble, W.S., Elferink, L.A., Wendland, B., Miller, K., Calakos, N., Scheller, R.H. (1990). Molecular Analysis of Proteins Associated with the Synaptic Vesicle Membrane. Cold Spring Harb Symp Quant Biol 55: 111-118 [Abstract]  
• Kelly, R.B., Buckley, K.M., Burgess, T.L., Carlson, S.S., Caroni, P., Hooper, J.E., Katzen, A., Moore, H.-P., Pfeffer, S.R., Schroer, T.A. (1983). Membrane Traffic in Neurons and Peptide-secreting Cells. Cold Spring Harb Symp Quant Biol 48: 697-705 [Abstract]  

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