Correlative Light-Electron Microscopy Reveals the Tubular-Saccular Ultrastructure of Carriers Operating between Golgi Apparatus and Plasma Membrane

Transport intermediates (TIs) have a central role in intracellular traffic, and much effort has been directed towards defining their molecular organization. Unfortunately, major uncertainties remain regarding their true structure in living cells. To address this question, we have developed an approa...

Full description

Saved in:
Bibliographic Details
Published in:The Journal of cell biology 2000-01, Vol.148 (1), p.45-58
Main Authors: Polishchuk, Roman S., Polishchuk, Elena V., Marra, Pierfrancesco, Alberti, Saverio, Buccione, Roberto, Luini, Alberto, Mironov, Alexander A.
Format: Article
Language:English
Subjects:
Animals
Antibodies
Cell Membrane - metabolism
Cell Membrane - ultrastructure
Cell membranes
Cells
Cellular biology
Chromaffin cells
Correlatives
COS Cells
Fluorescence
Freight
Golgi apparatus
Golgi Apparatus - metabolism
Golgi Apparatus - ultrastructure
Humans
Membrane Glycoproteins
Membranes
Microscopy
Microscopy, Electron - methods
Microtomy
Original
Proteins
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - metabolism
Somatic cells
Viral Envelope Proteins - genetics
Viral Envelope Proteins - metabolism
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Transport intermediates (TIs) have a central role in intracellular traffic, and much effort has been directed towards defining their molecular organization. Unfortunately, major uncertainties remain regarding their true structure in living cells. To address this question, we have developed an approach based on the combination of the green fluorescent protein technology and correlative light-electron microscopy, by which it is possible to monitor an individual carrier in vivo and then take a picture of its ultrastructure at any moment of its lifecycle. We have applied this technique to define the structure of TIs operating from the Golgi apparatus to the plasma membrane, whose in vivo dynamics have been characterized recently by light microscopy. We find that these carriers are large (ranging from 0.3-1.7 μm in maximum diameter, nearly half the size of a Golgi cisterna), comprise almost exclusively tubular-saccular structures, and fuse directly with the plasma membrane, sometimes minutes after docking to the fusion site.
ISSN:0021-9525
1540-8140
DOI:10.1083/jcb.148.1.45