Anastral meiotic spindles are thought to be organized differently from astral mitotic spindles, but the field lacks the basic structural information required to describe and model them, including the location of microtubule-nucleating sites and minus ends. We measured the distributions of oriented microtubules in metaphase anastral spindles in Xenopus laevis extracts by fluorescence speckle microscopy and cross-correlation analysis. We localized plus ends by tubulin incorporation and combined this with the orientation data to infer the localization of minus ends. We found that minus ends are localized throughout the spindle, sparsely at the equator and at higher concentrations near the poles. Based on these data, we propose a model for maintenance of the metaphase steady-state that depends on continuous nucleation of microtubules near chromatin, followed by sorting and outward transport of stabilized minus ends, and, eventually, their loss near poles.
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6 November 2006
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November 06 2006
A new method reveals microtubule minus ends throughout the meiotic spindle
Kendra S. Burbank,
Kendra S. Burbank
1Department of Physics, Harvard University, Cambridge, MA 03138
2Department of Systems Biology, Harvard Medical School, Boston, MA 02115
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Aaron C. Groen,
Aaron C. Groen
2Department of Systems Biology, Harvard Medical School, Boston, MA 02115
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Zachary E. Perlman,
Zachary E. Perlman
2Department of Systems Biology, Harvard Medical School, Boston, MA 02115
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Daniel S. Fisher,
Daniel S. Fisher
1Department of Physics, Harvard University, Cambridge, MA 03138
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Timothy J. Mitchison
Timothy J. Mitchison
2Department of Systems Biology, Harvard Medical School, Boston, MA 02115
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Kendra S. Burbank
1Department of Physics, Harvard University, Cambridge, MA 03138
2Department of Systems Biology, Harvard Medical School, Boston, MA 02115
Aaron C. Groen
2Department of Systems Biology, Harvard Medical School, Boston, MA 02115
Zachary E. Perlman
2Department of Systems Biology, Harvard Medical School, Boston, MA 02115
Daniel S. Fisher
1Department of Physics, Harvard University, Cambridge, MA 03138
Timothy J. Mitchison
2Department of Systems Biology, Harvard Medical School, Boston, MA 02115
Correspondence to Kendra S. Burbank: [email protected]
Abbreviation used in this paper: MT, microtubule.
Received:
November 28 2005
Accepted:
September 29 2006
Online ISSN: 1540-8140
Print ISSN: 0021-9525
The Rockefeller University Press
2006
J Cell Biol (2006) 175 (3): 369–375.
Article history
Received:
November 28 2005
Accepted:
September 29 2006
Citation
Kendra S. Burbank, Aaron C. Groen, Zachary E. Perlman, Daniel S. Fisher, Timothy J. Mitchison; A new method reveals microtubule minus ends throughout the meiotic spindle . J Cell Biol 6 November 2006; 175 (3): 369–375. doi: https://doi.org/10.1083/jcb.200511112
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