Actin dynamics provides membrane tension to merge fusing vesicles into the plasma membrane

Nat Commun. 2016 Aug 31:7:12604. doi: 10.1038/ncomms12604.

Abstract

Vesicle fusion is executed via formation of an Ω-shaped structure (Ω-profile), followed by closure (kiss-and-run) or merging of the Ω-profile into the plasma membrane (full fusion). Although Ω-profile closure limits release but recycles vesicles economically, Ω-profile merging facilitates release but couples to classical endocytosis for recycling. Despite its crucial role in determining exocytosis/endocytosis modes, how Ω-profile merging is mediated is poorly understood in endocrine cells and neurons containing small ∼30-300 nm vesicles. Here, using confocal and super-resolution STED imaging, force measurements, pharmacology and gene knockout, we show that dynamic assembly of filamentous actin, involving ATP hydrolysis, N-WASP and formin, mediates Ω-profile merging by providing sufficient plasma membrane tension to shrink the Ω-profile in neuroendocrine chromaffin cells containing ∼300 nm vesicles. Actin-directed compounds also induce Ω-profile accumulation at lamprey synaptic active zones, suggesting that actin may mediate Ω-profile merging at synapses. These results uncover molecular and biophysical mechanisms underlying Ω-profile merging.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism*
  • Animals
  • Cattle
  • Cell Membrane / metabolism*
  • Chromaffin Cells
  • Endocytosis
  • Exocytosis
  • Female
  • Gene Knockout Techniques
  • Image Processing, Computer-Assisted
  • Lampreys / genetics
  • Male
  • Membrane Fusion*
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Microscopy / methods
  • Models, Biological*
  • Molecular Imaging / methods
  • Neurons / metabolism
  • Patch-Clamp Techniques
  • Primary Cell Culture
  • Secretory Vesicles / metabolism
  • Synapses / metabolism
  • Synaptic Vesicles / metabolism

Substances

  • Actins