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

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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.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > Institute of Anatomy

UniBE Contributor:

Zuber, Benoît

Subjects:

600 Technology > 610 Medicine & health

ISSN:

0248-4900

Publisher:

Wiley

Language:

English

Submitter:

Benoît Zuber

Date Deposited:

28 Jan 2016 15:17

Last Modified:

28 Jan 2016 15:17

Publisher DOI:

10.1042/BC20060081

PubMed ID:

17049046

BORIS DOI:

10.7892/boris.74799

URI:

https://boris.unibe.ch/id/eprint/74799

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