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The Journal of Neuroscience, October 1, 1998, 18(19):7662-7673
Morphologically Docked Synaptic Vesicles Are Reduced in
synaptotagmin Mutants of Drosophila
Noreen E.
Reist1,
JoAnn
Buchanan2,
Jing
Li2,
Aaron
DiAntonio2,
Elizabeth M.
Buxton1, and
Thomas L.
Schwarz2
1 Department of Anatomy and Neurobiology, Colorado
State University, Fort Collins, Colorado 80523, and
2 Department of Molecular and Cellular Physiology, Stanford
University School of Medicine, Stanford, California 94305
Nerve terminal specializations include mechanisms for maintaining a
subpopulation of vesicles in a docked, fusion-ready state. We have
investigated the relationship between synaptotagmin and the number of
morphologically docked vesicles by an electron microscopic analysis of
Drosophila synaptotagmin (syt) mutants.
The overall number of synaptic vesicles in a terminal was reduced,
although each active zone continued to have a cluster of vesicles in
its vicinity. In addition, there was an increase in the number of large
vesicles near synapses. Examining the clusters, we found that the pool
of synaptic vesicles immediately adjacent to the presynaptic membrane,
the pool that includes the docked population, was reduced to 24 ± 5% (means ± SEM) of control in the
sytnull mutation.
To separate contributions of overall vesicle depletion and increased
spontaneous release from direct effects of synaptotagmin on
morphological docking, we examined syt mutants in an
altered genetic background. Recombining syt alleles onto
a second chromosome bearing an as yet uncharacterized mutation resulted
in the expected decrease in evoked release but suppressed the increase
in spontaneous release frequency. Motor nerve terminals in this
genotype contained more synaptic vesicles than control, yet the number
of vesicles immediately adjacent to the presynaptic membrane near
active zones was still reduced (33 ± 4% of control).
Our findings demonstrate that there is a decrease in the number of
morphologically docked vesicles seen in syt mutants. The decreases in docking and evoked release are independent of the increase
in spontaneous release. These results support the hypothesis that
synaptotagmin stabilizes the docked state.
Key words:
synaptic vesicles; Drosophila; synaptotagmin; electron microscopy; vesicle docking; vesicle
recycling
Copyright © 1998 Society for Neuroscience 0270-6474/98/18197662-12$05.00/0
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