Detection of mitotic spindle

Due to its essential role in cell division, the mitotic spindle has become an important therapeutic target for anticancer agents such as the Vinca alkaloids or taxanes. However, various mechanisms of developed resistance cause that these drugs often fail to induce desired mitotic block and death of cancer cells. Further investigation of microtubular system, namely full understanding its assembly, dynamics and regulation, is thus an incentive goal both for clinical and basic researchers.
 
In animal cells, a center of microtubule organization is the centrosome composed of a pair of cylindrical centrioles surrounded by fibrous pericentriolar material. Microtubules are nucleated from γ-tubulin ring structures embedded in the pericentriolar material. Formation of mitotic spindle is preceded by duplication of centrosome during S phase. Before mitosis, both centrosomes increase their microtubule nucleation capacity and form two microtubule asters that are pushed apart from each other by the forces of motor proteins associated at the microtubule surface. Upon nuclear envelope breakdown, formation of spindle apparatus is finalized by binding chromosomal kinetochores to centrosomal microtubules.
 
Fig. 1: Microtubules of mitotic spindle (α/β tubulin, green) originate from γ-tubulin rings (red) localized to pericentriolar material of centrosomes.




Fig. 2: Immunocytochemistry staining of tubulin (α/β-tubulin dimers) in HeLa cells using mouse monoclonal antibody TU-10 Alexa Fluor® 488. Mitotic spindle is visible even without detection of DNA.
 

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