A comparative analysis of methods to measure kinetochore-microtubule attachment stability. 2020

Jessica D Warren, and Bernardo Orr, and Duane A Compton
Department of Biochemistry and Cell Biology, Geisel School of Medicine, Hanover, NH, United States; Norris Cotton Cancer Center, Lebanon, NH, United States.

During mitosis, spindle microtubules dynamically attach to and detach from kinetochores in a precise and regulated fashion. To ensure mitotic fidelity, kinetochore-microtubule (k-MT) attachments must be stable enough to satisfy the spindle assembly checkpoint (SAC), but sufficiently unstable to facilitate the correction of maloriented attachments. Different methods are available to assess k-MT stability in both live and fixed cells, but a comparative survey of these methods has not yet been reported. Here, we evaluate several quantitative and semiquantitative methods for determining k-MT stability and apply each technique to illustrate changes in spindle microtubule dynamics upon perturbation with physiologically relevant concentrations of microtubule stabilizing (Taxol) and destabilizing (UMK57 and nocodazole) compounds. We discuss the utility of each technique for defining specific features of spindle microtubule dynamics and k-MT attachment stability.

UI MeSH Term Description Entries
D008027 Light That portion of the electromagnetic spectrum in the visible, ultraviolet, and infrared range. Light, Visible,Photoradiation,Radiation, Visible,Visible Radiation,Photoradiations,Radiations, Visible,Visible Light,Visible Radiations
D008870 Microtubules Slender, cylindrical filaments found in the cytoskeleton of plant and animal cells. They are composed of the protein TUBULIN and are influenced by TUBULIN MODULATORS. Microtubule
D008941 Spindle Apparatus A microtubule structure that forms during CELL DIVISION. It consists of two SPINDLE POLES, and sets of MICROTUBULES that may include the astral microtubules, the polar microtubules, and the kinetochore microtubules. Mitotic Apparatus,Mitotic Spindle Apparatus,Spindle Apparatus, Mitotic,Meiotic Spindle,Meiotic Spindle Apparatus,Mitotic Spindle,Apparatus, Meiotic Spindle,Apparatus, Mitotic,Apparatus, Mitotic Spindle,Apparatus, Spindle,Meiotic Spindles,Mitotic Spindles,Spindle Apparatus, Meiotic,Spindle, Meiotic,Spindle, Mitotic,Spindles, Meiotic,Spindles, Mitotic
D002118 Calcium A basic element found in nearly all tissues. It is a member of the alkaline earth family of metals with the atomic symbol Ca, atomic number 20, and atomic weight 40. Calcium is the most abundant mineral in the body and combines with phosphorus to form calcium phosphate in the bones and teeth. It is essential for the normal functioning of nerves and muscles and plays a role in blood coagulation (as factor IV) and in many enzymatic processes. Coagulation Factor IV,Factor IV,Blood Coagulation Factor IV,Calcium-40,Calcium 40,Factor IV, Coagulation
D002460 Cell Line Established cell cultures that have the potential to propagate indefinitely. Cell Lines,Line, Cell,Lines, Cell
D003584 Cytological Techniques Methods used to study CELLS. Cytologic Technics,Cytological Technic,Cytological Technics,Cytological Technique,Technic, Cytological,Technics, Cytological,Technique, Cytological,Techniques, Cytological,Cytologic Technic,Technic, Cytologic,Technics, Cytologic
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D015739 Nocodazole Nocodazole is an antineoplastic agent which exerts its effect by depolymerizing microtubules. NSC-238159,Oncodazole,R-17934,R17934,NSC 238159,NSC238159,R 17934
D058639 Cold-Shock Response A sequence of responses that occur when an organism is exposed to excessive cold. In humans, a fall in skin temperature triggers gasping, hypertension, and hyperventilation. Cold Shock,Cold Shock Stress,Cold Stress,Cold-Stress Reaction,Cold-Stress Response,Cold Shock Response,Cold Shock Stresses,Cold Shocks,Cold Stress Reaction,Cold Stress Response,Cold-Shock Responses,Cold-Stress Reactions,Cold-Stress Responses,Response, Cold-Stress,Responses, Cold-Stress,Shock Stress, Cold,Shock Stresses, Cold,Stress, Cold,Stress, Cold Shock,Stresses, Cold Shock
D018386 Kinetochores Large multiprotein complexes that bind the centromeres of the chromosomes to the microtubules of the mitotic spindle during metaphase in the cell cycle. Kinetochore

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