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Journal of Neuroscience, Vol 11, 1032-1039, Copyright © 1991 by Society for Neuroscience
Distribution of Ca2+ channels on frog motor nerve terminals revealed by fluorescent omega-conotoxin
MW Cohen, OT Jones and KJ Angelides
Department of Physiology, McGill University, Montreal, Quebec, Canada.
Tetramethylrhodamine-conjugated omega-conotoxin was used as a fluorescent
stain (Jones et al., 1989) to determine the spatial distribution of
voltage-gated Ca2+ channels along frog motor nerve terminals. Like native
omega-conotoxin, the fluorescent toxin blocked neuromuscular transmission
irreversibly. The fluorescent staining was confined to the neuromuscular
junction and consisted of a series of narrow bands (in face views) or dots
(in side views) approximately 1 micron apart. This characteristic staining
pattern was prevented by pretreatment with omega-conotoxin and by prior
denervation for 5-7 d. Combined fluorescence and phase-contrast optics
indicated that the stain was on the synaptic rather than the nonsynaptic
side of the nerve terminal. The bands and dots of stain proved to be in
spatial register with the postsynaptic junctional folds, as revealed by
combined staining of ACh receptors. It is concluded that the voltage-gated
Ca2+ channels on frog motor nerve terminals are concentrated at active
zones. The findings are consistent with the suggestion (Heuser et al.,
1974; Pumplin et al., 1981) that the large intramembraneous particles seen
at freeze-fractured active zones are voltage-gated Ca2+ channels.
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