Visualization of cell microtubules in their native state.
Details
Serval ID
serval:BIB_188E77703C00
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Visualization of cell microtubules in their native state.
Journal
Biology of the Cell
ISSN
0248-4900
Publication state
Published
Issued date
2007
Peer-reviewed
Oui
Volume
99
Number
1
Pages
45-53
Language
english
Notes
Journal Article --- Old month value: Jan
Abstract
BACKGROUND INFORMATION: Over the past decades, cryo-electron microscopy of vitrified specimens has yielded a detailed understanding of the tubulin and microtubule structures of samples reassembled in vitro from purified components. However, our knowledge of microtubule structure in vivo remains limited by the chemical treatments commonly used to observe cellular architecture using electron microscopy. RESULTS: We used cryo-electron microscopy and cryo-electron tomography of vitreous sections to investigate the ultrastructure of microtubules in their cellular context. Vitreous sections were obtained from organotypic slices of rat hippocampus and from Chinese-hamster ovary cells in culture. Microtubules revealed their protofilament ultrastructure, polarity and, in the most favourable cases, molecular details comparable with those visualized in three-dimensional reconstructions of microtubules reassembled in vitro from purified tubulin. The resolution of the tomograms was estimated to be approx. 4 nm, which enabled the detection of luminal particles of approx. 6 nm in diameter inside microtubules. CONCLUSIONS: The present study provides a first step towards a description of microtubules, in addition to other macromolecular assemblies, in an unperturbed cellular context at the molecular level. As the resolution appears to be similar to that obtainable with plunge-frozen samples, it should allow for the in vivo identification of larger macromolecular assemblies in vitreous sections of whole cells and tissues.
Keywords
Animals, CHO Cells, Cell Polarity, Cricetinae, Cricetulus, Cryoelectron Microscopy, Hippocampus, Microtubules, Neurons, Rats
Pubmed
Web of science
Open Access
Yes
Create date
24/01/2008 11:25
Last modification date
20/08/2019 13:49