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Title: Laser ablation and fluid flows reveal the mechanism behind spindle and centrosome positioning
Few techniques are available for studying the nature of forces that drive subcellular dynamics. Here we develop two complementary ones. The first is femtosecond stereotactic laser ablation, which rapidly creates complex cuts of subcellular structures and enables precise dissection of when, where and in what direction forces are generated. The second is an assessment of subcellular fluid flows by comparison of direct flow measurements using microinjected fluorescent nanodiamonds with large-scale fluid-structure simulations of different force transduction models. We apply these techniques to study spindle and centrosome positioning in early Caenorhabditis elegans embryos and to probe the contributions of microtubule pushing, cytoplasmic pulling and cortical pulling upon centrosomal microtubules. Based on our results, we construct a biophysical model to explain the dynamics of centrosomes. We demonstrate that cortical pulling forces provide a general explanation for many behaviours mediated by centrosomes, including pronuclear migration and centration, rotation, metaphase spindle positioning, asymmetric spindle elongation and spindle oscillations. This work establishes methodologies for disentangling the forces responsible for cell biological phenomena.  more » « less
Award ID(s):
1944156
NSF-PAR ID:
10318639
Author(s) / Creator(s):
; ; ; ; ;
Publisher / Repository:
Nature
Date Published:
Journal Name:
Nature physics
ISSN:
1745-2481
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
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