The mouth of a dense-core vesicle opens and closes in a concerted action regulated by calcium and amphiphysin.
Secretion of hormones and peptides by neuroendocrine cells occurs through fast and slow modes of vesicle fusion but the mechanics of these processes are not understood. We used interference reflection microscopy to monitor deformations of the membrane surface and found that both modes of fusion involve the tightly coupled dilation and constriction of the vesicle. The rate of opening is calcium dependent and occurs rapidly at concentrations <5 muM [corrected] The fast mode of fusion is blocked selectively by a truncation mutant of amphiphysin. Vesicles do not collapse when fusion is triggered by strontium, rather they remain locked open and membrane scission is blocked. In contrast, constriction of the vesicle opening continues when endocytosis is blocked by inhibiting the function of dynamin. Thus, fast and slow modes of fusion involve similar membrane deformations and vesicle closure can be uncoupled from membrane scission. Regulation of these processes by calcium and amphiphysin may provide a mechanism for controlling the release of vesicle contents.
Llobet
Artur
A
Medical Research Council Laboratory of Molecular Biology, Cambridge, England, UK. allobet@ub.edu
Wu
Minnie
M
Lagnado
Leon
L
D000818:Animals; D002118:Calcium / Q000378:metabolism / Q000502:physiology*; D002417:Cattle; D002478:Cells, Cultured; D034281:Dynamins / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure; D008561:Membrane Fusion / Q000502:physiology*; D008857:Microscopy, Interference; D009419:Nerve Tissue Proteins / Q000378:metabolism / Q000502:physiology*; D022142:Secretory Vesicles / Q000502:physiology* / Q000648:ultrastructure
D009419:Nerve Tissue Proteins; C075637:amphiphysin; D034281:Dynamins; D002118:Calcium
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