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Mol Biol Cell. 2013 May;24(9):1375-86. doi: 10.1091/mbc.E12-12-0901. Epub 2013 Mar 13.

Measurements of forces produced by the mitotic spindle using optical tweezers.

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1
Department of Biology, York University, Toronto, ON M3J 1P3, Canada. ferraroj@yorku.ca

Abstract

We used a trapping laser to stop chromosome movements in Mesostoma and crane-fly spermatocytes and inward movements of spindle poles after laser cuts across Potorous tridactylus (rat kangaroo) kidney (PtK2) cell half-spindles. Mesostoma spermatocyte kinetochores execute oscillatory movements to and away from the spindle pole for 1-2 h, so we could trap kinetochores multiple times in the same spermatocyte. The trap was focused to a single point using a 63× oil immersion objective. Trap powers of 15-23 mW caused kinetochore oscillations to stop or decrease. Kinetochore oscillations resumed when the trap was released. In crane-fly spermatocytes trap powers of 56-85 mW stopped or slowed poleward chromosome movement. In PtK2 cells 8-mW trap power stopped the spindle pole from moving toward the equator. Forces in the traps were calculated using the equation F = Q'P/c, where P is the laser power and c is the speed of light. Use of appropriate Q' coefficients gave the forces for stopping pole movements as 0.3-2.3 pN and for stopping chromosome movements in Mesostoma spermatocytes and crane-fly spermatocytes as 2-3 and 6-10 pN, respectively. These forces are close to theoretical calculations of forces causing chromosome movements but 100 times lower than the 700 pN measured previously in grasshopper spermatocytes.

PMID:
23485565
PMCID:
PMC3639049
DOI:
10.1091/mbc.E12-12-0901
[Indexed for MEDLINE]
Free PMC Article
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