Format

Send to

Choose Destination
Cell. 2018 May 3;173(4):934-945.e12. doi: 10.1016/j.cell.2018.02.062. Epub 2018 Apr 5.

Visualization of Membrane Pore in Live Cells Reveals a Dynamic-Pore Theory Governing Fusion and Endocytosis.

Author information

1
National Institute of Neurological Disorders and Stroke, Bethesda, MD, USA.
2
National Institute of Neurological Disorders and Stroke, Bethesda, MD, USA. Electronic address: wul@ninds.nih.gov.

Abstract

Fusion is thought to open a pore to release vesicular cargoes vital for many biological processes, including exocytosis, intracellular trafficking, fertilization, and viral entry. However, fusion pores have not been observed and thus proved in live cells. Its regulatory mechanisms and functions remain poorly understood. With super-resolution STED microscopy, we observed dynamic fusion pore behaviors in live (neuroendocrine) cells, including opening, expansion, constriction, and closure, where pore size may vary between 0 and 490 nm within 26 milliseconds to seconds (vesicle size: 180-720 nm). These pore dynamics crucially determine the efficiency of vesicular cargo release and vesicle retrieval. They are generated by competition between pore expansion and constriction. Pharmacology and mutation experiments suggest that expansion and constriction are mediated by F-actin-dependent membrane tension and calcium/dynamin, respectively. These findings provide the missing live-cell evidence, proving the fusion-pore hypothesis, and establish a live-cell dynamic-pore theory accounting for fusion, fission, and their regulation.

KEYWORDS:

STED; chromaffin cell; endocytosis; exocytosis; fission; fusion; fusion pore; pore constriction; pore expansion; release; super-resolution imaging

PMID:
29606354
PMCID:
PMC5935532
[Available on 2019-05-03]
DOI:
10.1016/j.cell.2018.02.062

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center