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Co-movement of astral microtubules, organelles and F-actin suggests aster positioning by surface forces in frog eggs

James Pelletier, Christine Field, Sebastian Fürthauer, Matthew Sonnett, Timothy Mitchison
doi: https://doi.org/10.1101/2020.06.17.154260
James Pelletier
1Harvard Medical School, Department of Systems Biology, Boston, MA 02115
2Marine Biological Laboratory, Woods Hole, MA 02543
3Massachusetts Institute of Technology, Department of Physics, Cambridge, MA 02139
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Christine Field
1Harvard Medical School, Department of Systems Biology, Boston, MA 02115
2Marine Biological Laboratory, Woods Hole, MA 02543
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Sebastian Fürthauer
4Flatiron Institute, Center for Computational Biology, New York, NY 10010
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Matthew Sonnett
1Harvard Medical School, Department of Systems Biology, Boston, MA 02115
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Timothy Mitchison
1Harvard Medical School, Department of Systems Biology, Boston, MA 02115
2Marine Biological Laboratory, Woods Hole, MA 02543
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  • For correspondence: timothy_mitchison@hms.harvard.edu
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Abstract

How bulk cytoplasm generates forces to separate post-anaphase microtubule (MT) asters in Xenopus laevis and other large eggs remains unclear. Previous models proposed dynein-based organelle transport generates length-dependent forces on astral MTs that pull centrosomes through the cytoplasm, away from the midplane. In Xenopus egg extracts, we co-imaged MTs, endoplasmic reticulum (ER), mitochondria, acidic organelles, F-actin, keratin, and fluorescein in moving and stationary asters. In asters that were moving in response to dynein and actomyosin forces, we observed that all cytoplasmic components moved together, i.e., as a continuum. Dynein-mediated organelle transport was restricted by interior MTs and F-actin. Organelles exhibited a burst of dynein-dependent inward movement at the growing aster surface, then mostly halted inside the aster. Dynein-coated beads were slowed by F-actin, but in contrast to organelles, beads did not halt inside asters. These observations call for new models of aster positioning based on surface forces and internal stresses.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted June 17, 2020.
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Co-movement of astral microtubules, organelles and F-actin suggests aster positioning by surface forces in frog eggs
James Pelletier, Christine Field, Sebastian Fürthauer, Matthew Sonnett, Timothy Mitchison
bioRxiv 2020.06.17.154260; doi: https://doi.org/10.1101/2020.06.17.154260
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Co-movement of astral microtubules, organelles and F-actin suggests aster positioning by surface forces in frog eggs
James Pelletier, Christine Field, Sebastian Fürthauer, Matthew Sonnett, Timothy Mitchison
bioRxiv 2020.06.17.154260; doi: https://doi.org/10.1101/2020.06.17.154260

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