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Dynamic Spatiotemporal Organization of Exocytosis During Cellular Shape Change

Fabio Urbina, Shawn Gomez, Stephanie L. Gupton
doi: https://doi.org/10.1101/185249
Fabio Urbina
1Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA
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Shawn Gomez
2UNC/NCSU Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill/North Carolina State University, Chapel Hill, NC 27517, USA
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Stephanie L. Gupton
1Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA
3Neuroscience Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA
4Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA
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Abstract

Morphological elongation of developing neurons requires plasmalemma expansion, hypothesized to occur primarily via exocytosis. We posited that exocytosis in developing neurons and non-neuronal cells exhibit distinct spatiotemporal organization. We exploited TIRF microscopy to image VAMP-pHluorin mediated exocytosis in murine embryonic cortical neurons and interphase melanoma cells, and developed computer-vision software and statistical tools to uncover spatiotemporal aspects of exocytosis. Vesicle fusion behavior differed between vesicle types, cell types, developmental stage, and extracellular environment. Experiment-based mathematical calculations indicated that VAMP2-mediated vesicle fusion supplied excess material for the plasma membrane expansion that occurred early in neuronal morphogenesis, which was balanced in part by clathrin-mediated endocytosis. Spatial statistics uncovered distinct spatiotemporal regulation of exocytosis in the soma and neurites of developing neurons that was modulated by developmental stage, exposure to the guidance cue netrin-1, and the brain-enriched ubiquitin ligase TRIM9. In melanoma cells, exocytosis occurred less frequently with distinct spatial clustering patterns.

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Posted September 06, 2017.
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Dynamic Spatiotemporal Organization of Exocytosis During Cellular Shape Change
Fabio Urbina, Shawn Gomez, Stephanie L. Gupton
bioRxiv 185249; doi: https://doi.org/10.1101/185249
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Dynamic Spatiotemporal Organization of Exocytosis During Cellular Shape Change
Fabio Urbina, Shawn Gomez, Stephanie L. Gupton
bioRxiv 185249; doi: https://doi.org/10.1101/185249

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