Dissecting docking and tethering of secretory vesicles at the target membrane

Research output: Contribution to journalJournal articleResearchpeer-review

  • Ruud F Toonen
  • Olexiy Kochubey
  • Heidi de Wit
  • Attila Gulyas-Kovacs
  • Bas Konijnenburg
  • Sørensen, Jakob Balslev
  • Jurgen Klingauf
  • Matthijs Verhage
Secretory vesicles dock at their target in preparation for fusion. Using single-vesicle total internal reflection fluorescence microscopy in chromaffin cells, we show that most approaching vesicles dock only transiently, but that some are captured by at least two different tethering modes, weak and strong. Both vesicle delivery and tethering depend on Munc18-1, a known docking factor. By decreasing the amount of cortical actin by Latrunculin A application, morphological docking can be restored artificially in docking-deficient munc18-1 null cells, but neither strong tethering nor fusion, demonstrating that morphological docking is not sufficient for secretion. Deletion of the t-SNARE and Munc18-1 binding partner syntaxin, but not the v-SNARE synaptobrevin/VAMP, also reduces strong tethering and fusion. We conclude that docking vesicles either undock immediately or are captured by minimal tethering machinery and converted in a munc18-1/syntaxin-dependent, strongly tethered, fusion-competent state.
Original languageEnglish
JournalEMBO Journal
Volume25
Issue number16
Pages (from-to)3725-37
Number of pages12
ISSN0261-4189
DOIs
Publication statusPublished - 2006
Externally publishedYes

Bibliographical note

Keywords: Actins; Animals; Cell Membrane; Cells, Cultured; Chromaffin Cells; Exocytosis; Gene Dosage; Membrane Fusion; Mice; Mice, Knockout; Microscopy, Electron, Transmission; Munc18 Proteins; Qa-SNARE Proteins; R-SNARE Proteins; SNARE Proteins; Secretory Vesicles

ID: 16835434