Oblique circle method for measuring the curvature and twist of mitotic spindle microtubule bundles

Ivec, Arian; Trupinić, Monika; Tolić, Iva M.; Pavin, Nenad (2021) Oblique circle method for measuring the curvature and twist of mitotic spindle microtubule bundles. Biophysical journal, 120 (17). pp. 3641-3648. ISSN 0006-3495

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Abstract

The highly ordered spatial organization of microtubule bundles in the mitotic spindle is crucial for its proper functioning. The recent discovery of twisted shapes of microtubule bundles and spindle chirality suggests that the bundles extend along curved paths in three dimensions, rather than being confined to a plane. This, in turn, implies that rotational forces, i.e., torques, exist in the spindle in addition to the widely studied linear forces. However, studies of spindle architecture and forces are impeded by a lack of a robust method for the geometric quantification of microtubule bundles in the spindle. In this work, we describe a simple method for measuring and evaluating the shapes of microtubule bundles by characterizing them in terms of their curvature and twist. By using confocal microscopy, we obtain three-dimensional images of spindles, which allows us to trace the entire microtubule bundle. For each traced bundle, we first fit a plane and then fit a circle lying in that plane. With this robust method, we extract the curvature and twist, which represent the geometric information characteristic for each bundle. As the bundle shapes reflect the forces within them, this method is valuable for the understanding of forces that act on chromosomes during mitosis.

Item Type:	Article
Uncontrolled Keywords:	Mitotic spindle ; chirality ; twist ; helicity ; methods
Subjects:	NATURAL SCIENCES > Physics NATURAL SCIENCES > Biology NATURAL SCIENCES > Interdisciplinary Natural Sciences
Divisions:	Division of Molecular Biology
Projects:	Project title Project leader Project code Project type Ravnoteža sila i momenta sila u diobenom vretenu-SpindleForceBalance Nenad Pavin IP-2019-04-5967 HRZZ Molecular origins of aneuploidies in healthy and diseased human tissues-ANEUPLOIDY Iva Tolić; Nenad Pavin 855158 EK A new class of microtubules in the spindle exerting forces on kinetochores-NewSpindleForce Iva Tolić 647077 EK Mehanizmi nastajanja snopova mikrotubula potrebni za sazrijevanje diobenog vretena-SpindleForm Iva Tolić PZS-2019-02-7653 HRZZ
Depositing User:	Iva Tolić
Date Deposited:	09 Nov 2021 14:59
URI:	http://fulir.irb.hr/id/eprint/6602
DOI:	10.1016/j.bpj.2021.07.024

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