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Visualization of cell microtubules in their native state
Visualization of cell microtubules in their native state
7329
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.
Bouchet-Marquis C, Zuber B, Glynn AM, Eltsov M, Grabenbauer M, Goldie KN, Thomas D, Frangakis AS, Dubochet J, Chrétien D
Biology of the cell / under the auspices of the European Cell Biology Organization
2007-01-01 00:00
99
1
45-53
Animals,CHO Cells,Cell Polarity,Cricetinae,Cricetulus,Cryoelectron Microscopy,Hippocampus,Microtubules,Neurons,Rats
Laboratory for Ultrastructural Analysis, Biophore, University of Lausanne, CH-1015 Lausanne, Switzerland.
Biol. Cell
0248-4900
10.1042/BC20060081
BC20060081
0
False
17049046
