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  • 1
    ISSN: 1573-6822
    Keywords: cytoskeleton ; embryonal carcinoma cells ; immunofluorescence ; methylmercury ; mitosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunofuorescence staining with antibodies to tubulin and vimentin and staining with phalloidin have been used to examine the effects of methylmercury on the cytoskeleton of embryonal carcinoma cells in culture. Exposure of embryonal carcinoma cells to methylmercury (0.01 to 10 μm) resulted in concentration- and time-dependent disassembly of microtubules in interphase and mitotic cells. These effects were reversible when cultures were washed free of methylmercury. Spindle microtubules were more sensitive than those of interphase cells. Spindle damage resulted in an accumulation of cells in prometaphase/metaphase, which; correlated with a temporary delay in the resumption of normal proliferation rate upon removal of methylmercury. Of the interphase cytoskeletal components, microtubules were the first affected by methylmercury. Vimentin intermediate filaments appeared relatively insensitive to methylmercury, but showed a reorganization secondary to the microtubule disassembly. Actin microfilaments appeared unchanged in cells showing complete absence of microtubules. Our results 1) support previous reports suggesting that microtubules are a primary target of methylmercury, 2) document a differential sensitivity of mitotic and interphase microtubule systems and 3) demonstrate the relative insensitivities of other cytoskeletal components.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 6 (1986), S. 122-127 
    ISSN: 0886-1544
    Keywords: microtubules ; rootlets ; monoclonals ; immunofluorescence ; mitosis ; cytokinesis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We have used immunofluorescence staining with monoclonal antibodies to tubulin and to components of the flagellar basal apparatus to examine duplication of the basal apparatus and the microtubule system assembled from it during cell division in the quadriflagellate alga Polytomella. The monoclonal antitubulin, prepared against Polytomella flagellar axonemes, detects Polytomella and mammalian tubulin by immunoblotting. By immunofluorescence and immunogold electron microscopy, it detects all microtubular structures that have been described in Polytomella. One of the antibodies generated using the isolated basal apparatus as immunogen appears to stain four of the eight basal body rootlets and is used in this study to detect early stages in the duplication of the flagellar apparatus. A cytoplasmic microtubule system is present, the elongate morphology of the cell is maintained, and the cells are motile throughout mitosis. The closed mitotic spindle forms perpendicular to the long axis of the cell. During mitosis, the newly formed basal bodies mature and four additional elongating flagella appear. Following mitosis, the eight flagella segregate into two groups, which begin to separate towards opposite poles of the cell. Concomitant with this separation, the rootlets of the parental basal apparatus separate and new rootlets are detected. We suggest that the components of the parental flagellar apparatus are segregated equally to the daughter cells. An interphase cytoskeletal microtubule array is assembled from each basal apparatus, and the morphology of the two cells is progressively established during cytokinesis.
    Additional Material: 3 Ill.
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 8 (1987), S. 227-237 
    ISSN: 0886-1544
    Keywords: Vimentin ; tubulin ; lymphocytes ; stimulation ; mitosis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We have used double immunofluorescence and electron microscopy to examine the distribution of tubulin and vimentin during the stimulation of mouse splenic lymphocytes by the mitogen concanavalin A. In unstimulated cells, vimentin forms a filamentous network partially coincident with the radial pattern of microtubules. In stimulated cells, the numbers of microtubules assembled from the centrosome. When these cells enter mitosis, vimentin is arranged into a filamentous cage enclosing the mitotic apparatus. During cytokinesis, the polar centrosomes are observed at a position adjacent to the midbody and vimentin is detected as an aggregate, similar to that seen prior to mitosis, close to the centrosome in each daughter cell. Using several agents, such as colchicine, colcemid, nocodazole, and taxol, which affect microtubule assembly, we have observed that the vimentin system, although closely related spatially to the microtubule complex in lymphocytes, can still reorganize independently as these cells progress through in the cell cycle. Throughout mitogenic stimulation in the continued presence of taxol, microtubules are reorganized into a few thick bundles while the vimentin system undergoes a sequence of rearragements similar to those observed during normal stimulation. These data suggest that vimentin dynamics may be important in the progression of lymphocytes through the cell cycle in response to mitogen.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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