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Vol. 13, Issue 11, 3967-3975, November 2002
Center for Genetics and Development and Section of Molecular and
Cellular Biology, University of California, Davis, California 95616
We proposed that spindle morphogenesis in Drosophila
embryos involves progression through four transient isometric
structures in which a constant spacing of the spindle poles is
maintained by a balance of forces generated by multiple microtubule
(MT) motors and that tipping this balance drives pole-pole
separation. Here we used fluorescent speckle microscopy to
evaluate the influence of MT dynamics on the isometric state that
persists through metaphase and anaphase A and on pole-pole separation
in anaphase B. During metaphase and anaphase A, fluorescent punctae on
kinetochore and interpolar MTs flux toward the poles at 0.03 µm/s,
too slow to drive chromatid-to-pole motion at 0.11 µm/s, and during
anaphase B, fluorescent punctae on interpolar MTs move away from the
spindle equator at the same rate as the poles, consistent with MT-MT
sliding. Loss of Ncd, a candidate flux motor or brake, did not affect
flux in the metaphase/anaphase A isometric state or MT sliding in
anaphase B but decreased the duration of the isometric state. Our
results suggest that, throughout this isometric state, an outward force exerted on the spindle poles by MT sliding motors is balanced by flux,
and that suppression of flux could tip the balance of forces at the
onset of anaphase B, allowing MT sliding and polymerization to push the
poles apart.
Online version contains video materials
for Figures 1 and 2. Online version is available at
www.molbiolcell.org.
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