Golgi apparatus studied in vitreous sections.
Details
Serval ID
serval:BIB_7CCB93A81DE4
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Golgi apparatus studied in vitreous sections.
Journal
Journal of microscopy
ISSN
1365-2818 (Electronic)
ISSN-L
0022-2720
Publication state
Published
Issued date
05/2008
Peer-reviewed
Oui
Volume
230
Number
Pt 2
Pages
308-316
Language
english
Notes
Publication types: Evaluation Study ; Journal Article
Publication Status: ppublish
Publication Status: ppublish
Abstract
Cryo-electron microscopy of vitrified specimen is the method of choice to explore cellular ultrastructure at high resolution as close as possible to the native state and environment. In this study, we investigated the Golgi apparatus - the main organelle of the secretory pathway. Cultured mammalian cells were fixed by high-pressure freezing, sectioned in vitreous ice and subjected to cryo-electron microscopy and cryo-electron tomography. Although the overall morphology of Golgi stacks was comparable to well prepared and plastic-embedded samples, in detail we reached much higher resolution in terms of distinction between biological structures based on their native density. On cisternal buds and peri-Golgi vesicles--some associated with microtubules--we detected two different subtypes of COPI coats: (1) a homogenous coat and (2) an inhomogeneous spiky coat, providing an 8-9 nm regularity, clearly distinct from clathrin coat. Next, we monitored the secretion of cargo, namely, procollagen I, through the Golgi complex. Temporally correlated with fluorescence microscopy, we performed three-dimensional cryo-electron tomography analysis and detected Golgi cisternae enlarged to saccules, containing cargo and showing inter-cisternal connections. Our work provides a first step towards the high-resolution description of the secretory pathway in native vitrified samples and describes the challenges associated with this attempt.
Keywords
Animals, CHO Cells/ultrastructure, COP-Coated Vesicles/ultrastructure, Cricetinae, Cricetulus, Cryoelectron Microscopy/methods, Cryopreservation/methods, Freezing, Frozen Sections, Golgi Apparatus/ultrastructure, Hydrostatic Pressure, Microscopy, Fluorescence, Tomography
Pubmed
Web of science
Open Access
Yes
Create date
29/01/2009 23:14
Last modification date
13/02/2024 8:23