Chromosome elasticity and mitotic polar ejection force measured in living Drosophila embryos by four-dimensional microscopy-based motion analysis.

Abstract

Mitosis involves the interaction of many different components, including chromatin, microtubules, and motor proteins. Dissecting the mechanics of mitosis requires methods of studying not just each component in isolation, but also the entire ensemble of components in its full complexity in genetically tractable model organisms. We have developed a mathematical framework for analyzing motion in four-dimensional microscopy data sets that allows us to measure elasticity, viscosity, and forces by tracking the conformational movements of mitotic chromosomes. We have used this approach to measure, for the first time, the basic biophysical parameters of mitosis in wild-type Drosophila melanogaster embryos. We found that Drosophila embryo chromosomes are significantly less rigid than the much larger chromosomes of vertebrates. Anaphase kinetochore force and nucleoplasmic viscosity were comparable with previous estimates in other species. Motion analysis also allowed us to measure the magnitude of the polar ejection force exerted on chromosome arms during metaphase by individual microtubules. We find the magnitude of this force to be approximately 1 pN, a number consistent with force generation either by collision of growing microtubules with chromosomes or by single kinesin motors. Motion analysis allows noninvasive mechanical measurements to be made in complex systems. This approach should allow the functional effects of Drosophila mitotic mutants on chromosome condensation, kinetochore forces, and the polar ejection force to be determined.