Analysis of metaphase spindle mechanics in HeLa cells by laser microsurgery

Polak, Bruno (2015) Analysis of metaphase spindle mechanics in HeLa cells by laser microsurgery. Diploma thesis, Faculty of Science > Department of Biology.

Language: English

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Main structure that orchestrates mitosis is the mitotic spindle, and one of the microtubule classes comprised within are the kinetochore fibers, which bind to chromosomes via kinetochores on the centromere. It is currently believed that there is no direct interaction between two sister kinetochore fibers. The focus of this research is to describe a novel class of microtubules in the spindle, which was named bridging microtubules. This new class forms antiparallel bundles, laterally connects sister kinetochore fibers and contributes to spindle mechanics. In conducted research, confocal microscope was used for live-cell imaging of HeLa, and the laser microsurgery was applied on outermost kinetochore fiber in order to confirm bridging microtubule presence and to test their function in force distribution in the spindle. By using cells with decreased level of PRC1 protein, protein involved in metaphase spindle mechanics, we showed that bridging bundle contributes to force balance in the spindle. Microtubule dynamics in the region between sister kinetochores was also analyzed in cells with labeled EB3 protein involved in the growth of microtubule 's + end. Laser microsurgery confirmed compressive and tensile force in the metaphase spindle in both experiments.

Item Type: Thesis (Diploma thesis)
Keywords: mitosis, microtubule, kinetochore, kinetochore fiber, ablation
Supervisor: Tolić, Iva
Co-supervisor: Matulić, Maja
Date: 2015
Number of Pages: 74
Subjects: NATURAL SCIENCES > Biology
Divisions: Faculty of Science > Department of Biology
Depositing User: Silvana Šehić
Date Deposited: 03 Mar 2016 13:40
Last Modified: 03 Mar 2016 13:40

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