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A more recent version of this article appeared on February 1, 2004
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Submitted on June 14, 2003
Revised on September 15, 2003
Accepted on October 11, 2003
1 Dept. of Cell Biology and The Center for Neurodegenerative Diseases. Emory University
2 Dept. of Cell Biology, Neurology, and The Center for Neurodegenerative Diseases. Emory University
3 The Center for Neurodegenerative Diseases. Emory University, Department of Cell Biology, Emory University, Whitehead Biomedical Research Bld., 615 Michael Street, Room 446, Atlanta, GA 30322
* Corresponding author. E-mail address: faundez{at}cellbio.emory.edu.
Synaptic vesicles (SV) are generated by two different mechanisms, one AP-2 dependent and one AP-3 dependent. It has been uncertain, however, whether these mechanisms generate SV that differ in molecular composition. We explored this hypothesis by analyzing the targeting of ZnT3 and synaptophysin both to PC12 synaptic-like microvesicles (SLMV) as well as synaptic vesicles (SV) isolated from wild-type and AP-3 deficient mocha brains. ZnT3 cytosolic tail interacted selectively with AP-3 in cell free assays. Accordingly, pharmacological disruption of either AP-2- or AP-3-dependent SLMV biogenesis preferentially reduced synaptophysin or ZnT3 targeting, respectively; suggesting that these antigens were concentrated in different vesicles. As predicted, immuno-isolated SLMV revealed that ZnT3 and synaptophysin were enriched in different vesicle populations. Likewise, morphological and biochemical analyses in hippocampal neurons indicated that these two antigens were also present in distinct but overlapping domains. ZnT3 SV content was reduced in AP-3 deficient neurons, but synaptophysin was not altered in the AP-3 null background. Our evidence indicates that neuroendocrine cells assemble molecularly heterogeneous SV, and suggest that this diversity could contribute to the functional variety of synapses.
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