Henderson, Gregory P. and Gan, Lu and Jensen, Grant J. (2007) 3-D Ultrastructure of O. tauri: Electron Cryotomography of an Entire Eukaryotic Cell. PLoS ONE, 2 (8). Art. No. e749. ISSN 1932-6203. PMCID PMC1939878. http://resolver.caltech.edu/CaltechAUTHORS:HENplosone07
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The hallmark of eukaryotic cells is their segregation of key biological functions into discrete, membrane-bound organelles. Creating accurate models of their ultrastructural complexity has been difficult in part because of the limited resolution of light microscopy and the artifact-prone nature of conventional electron microscopy. Here we explored the potential of the emerging technology electron cryotomography to produce three-dimensional images of an entire eukaryotic cell in a near-native state. Ostreococcus tauri was chosen as the specimen because as a unicellular picoplankton with just one copy of each organelle, it is the smallest known eukaryote and was therefore likely to yield the highest resolution images. Whole cells were imaged at various stages of the cell cycle, yielding 3-D reconstructions of complete chloroplasts, mitochondria, endoplasmic reticula, Golgi bodies, peroxisomes, microtubules, and putative ribosome distributions in-situ. Surprisingly, the nucleus was seen to open long before mitosis, and while one microtubule (or two in some predivisional cells) was consistently present, no mitotic spindle was ever observed, prompting speculation that a single microtubule might be sufficient to segregate multiple chromosomes.
|Additional Information:||© 2007 Henderson et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Received: May 21, 2007; Accepted: July 16, 2007; Published: August 15, 2007. We thank Hervé Moreau (Observatoire Océanologique, Laboratoire Arago) and Brian Palenik (Scripps Institution of Oceanography) for helping us obtain and work with O. tauri cultures and for reading the manuscript, Kent L. McDonald (U.C. Berkeley) for help high-pressure freezing, Wolfgang Baumeister's group (Max Planck Institute for Biochemistry) for the most current version of the MolMatch software, Andrea Manuell for discussions on the chloroplast, Christian Suloway (Caltech) for assistance with Leginon and Jane Ding (Caltech) for help running the cross-correlation search on the supercomputer. Author Contributions: Conceived and designed the experiments: GJ GH LG. Performed the experiments: GH LG. Analyzed the data: GH LG. Contributed reagents/materials/analysis tools: GJ. Wrote the paper: GJ GH LG. Funding: This work was supported in part by NIH grants R01 AI067548 and PO1 GM66521 and DOE grant DE-FG02-04ER63785 to GJJ, a Searle Scholar Award to GJJ, the Beckman Institute at Caltech, and gifts to Caltech from the Gordon and Betty Moore Foundation and Agouron Institute. Lu Gan was supported by a fellowship from the Damon Runyon Cancer Research Foundation (DRG-1940-07). These sponsors/funders had no role in the design and conduct of the study, in the collection, analysis, and interpretation of the data, and in the preparation, review, or approval of the manuscript. Competing interests: The authors have declared that no competing interests exist.|
|PubMed Central ID:||PMC1939878|
|Official Citation:||Henderson GP, Gan L, Jensen GJ (2007) 3-D Ultrastructure of O. tauri: Electron Cryotomography of an Entire Eukaryotic Cell. PLoS ONE 2(8): e749. doi:10.1371/journal.pone.0000749|
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|Deposited On:||09 Jan 2008|
|Last Modified:||13 May 2016 16:08|
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