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The Journal of Neuroscience, April 26, 2006, 26(17):4630-4637; doi:10.1523/JNEUROSCI.0009-06.2006

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Cellular/Molecular
A Role for Synaptotagmin VII-Regulated Exocytosis of Lysosomes in Neurite Outgrowth from Primary Sympathetic Neurons

Rosa M. E. Arantes1 and Norma W. Andrews1,2

1Section of Microbial Pathogenesis and 2Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06536

Correspondence should be addressed to Norma W. Andrews, Section of Microbial Pathogenesis, Boyer Center for Molecular Medicine, Yale University School of Medicine, 295 Congress Avenue, New Haven, CT 06510. Email: norma.andrews{at}yale.edu

Neurite outgrowth is mediated by the exocytosis of intracellular vesicles at the tips of elongating neuronal processes. The lysosomal vesicle-associated soluble N-ethylmaleimide-sensitive factor attachment protein receptor tetanus neurotoxin insensitive vesicle-associated membrane protein (TI-VAMP)/VAMP7 was previously implicated in membrane fusion events mediating neurite outgrowth, but the participation of lysosomes in this exocytic process has remained unclear. Here, we show that VAMP7 and the lysosomal glycoprotein Lamp1 extensively colocalize in vesicles present throughout the soma and neurite outgrowths of primary sympathetic neurons. Synaptotagmin VII (Syt VII), a Ca2+-sensing synaptotagmin isoform previously shown to interact with VAMP7 during lysosomal exocytosis in fibroblasts, was detected on a subset of these lysosomal glycoprotein 1 (Lamp1)/VAMP7-positive neuronal vesicles. Ionophore-stimulated exocytosis triggered exposure of the luminal domains of both Lamp1 and Syt VII at overlapping sites on the neuronal surface, indicating that the Syt VII-containing lysosomal compartments fuse with the plasma membrane in response to [Ca2+]i elevation. To determine whether Syt VII was required for the exocytic events mediating neurite extension, we followed the development of superior cervical ganglion neurons explanted from Syt VII-deficient mice. The results revealed a marked defect in neurite outgrowth and arborization, suggesting that Ca2+-dependent, Syt VII-regulated exocytosis of late endosomes/lysosomes plays a role in the addition of new membrane to developing neurite extensions.

Key words: calcium; dendrite; development; exocytosis; fluorescence microscopy; knock-out mice


Received Jan. 3, 2006; revised March 17, 2006; accepted March 22, 2006.

Correspondence should be addressed to Norma W. Andrews, Section of Microbial Pathogenesis, Boyer Center for Molecular Medicine, Yale University School of Medicine, 295 Congress Avenue, New Haven, CT 06510. Email: norma.andrews{at}yale.edu




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