Biomedical subjects

Alexey Khodjakov

Publications and source records attributed to Alexey Khodjakov.

23 records · Page 2Linked to original sources

PubMed2003

Mitosis through the microscope: advances in seeing inside live dividing cells.

The most visually spectacular events in the life of a cell occur when it divides. This is especially true in higher eukaryotes, where the size and geometry of cells allow the division process to be followed through a microscope with considerable clarity. In these organisms, the membrane surrounding the nucleus breaks down after the replicated DNA has condensed to form discrete chromosomes. Several new structures are then assembled to separate the chromosomes and partition the cytoplasm into two separate cells.

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PubMed2003

Minus-end capture of preformed kinetochore fibers contributes to spindle morphogenesis.

Near-simultaneous three-dimensional fluorescence/differential interference contrast microscopy was used to follow the behavior of microtubules and chromosomes in living alpha-tubulin/GFP-expressing cells after inhibition of the mitotic kinesin Eg5 with monastrol. Kinetochore fibers (K-fibers) were frequently observed forming in association with chromosomes both during monastrol treatment and after monastrol removal. Surprisingly, these K-fibers were oriented away from, and not directly connected to, centrosomes and incorporated into the spindle by the sliding of their distal ends toward centrosomes via a NuMA-dependent mechanism. Similar preformed K-fibers were also observed during spindle formation in untreated cells. In addition, upon monastrol removal, centrosomes established a transient chromosome-free bipolar array whose orientation specified the axis along which chromosomes segregated. We propose that the capture and incorporation of preformed K-fibers complements the microtubule plus-end capture mechanism and contributes to spindle formation in vertebrates.

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PubMed2002

De novo formation of centrosomes in vertebrate cells arrested during S phase.

The centrosome usually replicates in a semiconservative fashion, i.e., new centrioles form in association with preexisting "maternal" centrioles. De novo formation of centrioles has been reported for a few highly specialized cell types but it has not been seen in vertebrate somatic cells. We find that when centrosomes are completely destroyed by laser microsurgery in CHO cells arrested in S phase by hydroxyurea, new centrosomes form by de novo assembly. Formation of new centrosomes occurs in two steps: approximately 5-8 h after ablation, clouds of pericentriolar material (PCM) containing gamma-tubulin and pericentrin appear in the cell. By 24 h, centrioles have formed inside of already well-developed PCM clouds. This de novo pathway leads to the formation of a random number of centrioles (2-14 per cell). Although clouds of PCM consistently form even when microtubules are completely disassembled by nocodazole, the centrioles are not assembled under these conditions.

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PubMed2002

Dynamic microtubules in Dictyostelium.

The term 'microtubule dynamics' is often used to describe assembly/disassembly characteristics of this important cytoskeletal polymer. The ability to image microtubules in live Dictyostelium cells has revealed additional dynamic components, acting on the individual assembled tubules. At least two separate forces are involved, in generation of pronounced bending motions during interphase and in creating tension with the cell cortex. This review attempts to summarize what is known about conventional microtubule dynamics in Dictyostelium as well as to describe these two additional motility components. We propose that these forces are important both in maintaining the overall structure of the microtubule array and in supporting intracellular traffic.

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PubMed2002

Centrosome reorientation in wound-edge cells is cell type specific.

The reorientation of the microtubule organizing center during cell migration into a wound in the monolayer was directly observed in living wound-edge cells expressing gamma-tubulin tagged with green fluorescent protein. Our results demonstrate that in CHO cells, the centrosome reorients to a position in front of the nucleus, toward the wound edge, whereas in PtK cells, the centrosome lags behind the nucleus during migration into the wound. In CHO cells, the average rate of centrosome motion was faster than that of the nucleus; the converse was true in PtK cells. In both cell lines, centrosome motion was stochastic, with periods of rapid motion interspersed with periods of slower motion. Centrosome reorientation in CHO cells required dynamic microtubules and cytoplasmic dynein/dynactin activity and could be prevented by altering cell-to-cell or cell-to-substrate adhesion. Microtubule marking experiments using photoactivation of caged tubulin demonstrate that microtubules are transported in the direction of cell motility in both cell lines but that in PtK cells, microtubules move individually, whereas their movement is more coherent in CHO cells. Our data demonstrate that centrosome reorientation is not required for directed migration and that diverse cells use distinct mechanisms for remodeling the microtubule array during directed migration.

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