Modulating vesicle priming reveals that vesicle immobilization is necessary but not sufficient for fusion-competence

Ofer Yizhar*, Uri Ashery

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

In neurons and neuroendocrine cells, docked vesicles need to undergo priming to become fusion competent. Priming is a multi-step process that was shown to be associated with vesicle immobilization. However, it is not known whether vesicle immobilization is sufficient to acquire complete fusion competence. To extend our understanding of the physical manifestation of vesicle priming, we took advantage of tomosyn, a SNARE-related protein that specifically inhibits vesicle priming, and measured its effect on vesicle dynamics in live chromatin cells using total internal reflection fluorescence microscopy. We show here that while in control cells vesicles undergo immobilization before fusion, vesicle immobilization is attenuated in tomosyn overexpressing cells. This in turn increases the turnover rate of vesicles near the membrane and attenuates the fusion of newcomer vesicles. Moreover, the release probability of immobile vesicles in tomosyn cells is significantly reduced, suggesting that immobilization is an early and necessary step in priming but is insufficient, as further molecular processes are needed to acquire complete fusion competence. Using tomosyn as a molecular tool we provide a mechanistic link between functional docking and priming and suggest that functional docking is the first step in vesicle priming, followed by molecular modifications that do not translate into changes in vesicle mobility.

Original languageEnglish
Article numbere2694
JournalPLoS ONE
Volume3
Issue number7
DOIs
StatePublished - 16 Jul 2008

Funding

FundersFunder number
National Institute of Neurological Disorders and StrokeR01NS053978

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