Visualization of cell microtubules in their native state.

Bouchet-Marquis, Cédric; Zuber, Benoît; Glynn, Anne-Marie; Eltsov, Mikhail; Grabenbauer, Markus; Goldie, Kenneth N; Thomas, Daniel; Frangakis, Achilleas S; Dubochet, Jacques; Chrétien, Denis (2007). Visualization of cell microtubules in their native state. Biology of the cell, 99(1), pp. 45-53. Wiley 10.1042/BC20060081

Text 06.Bouchet-Marquis_Visualization_CellMicrotubules_2007.pdf - Published Version Restricted to registered users only Available under License Publisher holds Copyright. Download (933kB) | Request a copy

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.

Interest & Impact

Downloads

1 since deposited on 28 Jan 2016
0 in the past 12 months

Citations

5 Citations in Web of Science ®
6 Citations in Scopus

Search

in Google Scholar™

Services

Actions (login required)

Edit item

Actions (login required)

Edit item