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  • 1
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 447 (2007), S. 493-496 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] The architecture and adhesiveness of a cell microenvironment is a critical factor for the regulation of spindle orientation in vivo. Using a combination of theory and experiments, we have investigated spindle orientation in HeLa (human) cells. Here we show that spindle orientation can be ...
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  • 2
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] To determine whether the expression of Rab proteins varies during the cell cycle, whole cell extracts from interphase and mitotic HeLa cells were probed with specific polyclonal rabbit antisera raised against human Rab proteins expressed in Escherichia coll The levels of RablA, Rab2, Rab4 and Rab6 ...
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  • 3
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 355 (1992), S. 300-301 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] JUST over a century ago the centrosome was identified and dubbed the 'division organ' because of what seemed to be its main function at fertilization: organization of the first mitotic spindle of the embryo after duplication of the sperm centrosome. Although this picture has been shown to ...
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  • 4
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 295 (1982), S. 334-336 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] FCCP induced a disruption of HeLa cell microtubules (Fig. le and Table 1) which was complete after 1 h. The only tubulin-positive structures observed were midbodies and juxtanuclear micro tubule-organizing centres (MTOC). After 6 h treatment, short microtubules were seen regrowing from the MTOC. ...
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  • 5
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 270 (1977), S. 80-82 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Nuclei from rat liver were prepared according to Blobel and Potter5 with slight modifications to adapt the procedure to larger amounts of tissue. Liver tissue (33 g) was homogenised in 110ml of 2.2 M sucrose-TKM (50 mM Tris-HCl pH 7.5, 25 mM KC1, 5 mM MgCl2) with a Chauveau type homogeniser ...
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  • 6
    ISSN: 1432-0886
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract A rabbit serum which had previously been reported to have an immunological affinity for centrosomes of human cell lines was shown also to be specific for the nucleus. Optical and ultrastructural immunolocalization in HeLa cells showed that this specificity is restricted to the fibrillar centre of nucleoli either in untreated or actinomycin D treated interphase cells. In mitotic cells discrete labelling was observed on chromosomes and shown to correspond, on spread metaphase plates, to the short arms of acrocentric chromosomes, i.e. to the nucleolar organizer regions (NORs). Using independent cell fractionation procedures in the human T-lymphoblastic KE 37 cell line and purification of immunoglobulins by affinity to antigens detected by electrophoresis and blotting, a strict correlation between immunoreactive proteins and cytological staining was established. The nucleolar specificity was shown to correspond to a protein with an Mr of 80,000 while the centrosomal specificity corresponded principally to a protein doublet of 60,000–65,000. These antigens share common epitopes as shown by the staining of both NOR and centrosome by immunoglobulins purified by affinity to either type of protein.
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  • 7
    ISSN: 0886-1544
    Keywords: basal body ; centrosome ; ciliated epithelium ; ciliary rootlet ; cortex of metozoan ciliated cells ; monoclonal antibody ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Basal bodies from laying quail oviduct were semipurified and used as immunogen to produce monoclonal antibodies. On 38 clones obtained and among those staining the apical pole of the ciliated cell, CC-310 was chosen because it labeled the apical region with a punctated aspect, suggesting a staining of basal bodies or of basal body-associated structures; the basal pole was also labeled.The ultrastructural localization performed by the immunogold technique showed that the labeling was mainly associated with the striated rootlets. The basal feet, the side of the basal bodies, and the basal poles of the demembranated cells were also decorated. The identification of the antigen performed by immunoblots of deciliated cortices revealed two proteins of 175,000 and 40,000, whereas immunoblots of basal bodies showed only the 175,000-mw protein. The possibility of these two proteins sharing the same epitope, located at both poles of the cell, is discussed.Immunofluorescence ascertained that CC-310 decorated the striated rootlets in ciliated epithelia from other species: mussel, frog, and human tissue. Finally, when tested on cultured cell lines, CC-310 labeled the centrosome and its associated rootlets on PtK2 during interphase. During mitosis the poles of the mitotic spindle were stained without any apparent rootlet-like structure.
    Additional Material: 8 Ill.
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 23 (1992), S. 122-132 
    ISSN: 0886-1544
    Keywords: centrosome ; microtubule ; microfilament ; myogenesis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Available data on the molecular composition of the centrosome, the typical microtubule-organizing center of animal cells, are still fragmentary. To address this important issue we have taken advantage of centrosome isolation from a human lymphoblastic cell line (KE37) to generate a monoclonal antibody (mAb) library. Here we present the characterization of one of these mAbs (CTR56). On the basis of both its immunofluorescence staining pattern and its reactivity with a major 200 kD antigen on immunoblots, CTR56 has been tentatively classified as an anticellular myosin heavy chain. In light of cytological and biochemical data obtained in parallel with two other well-characterized myosin antibodies, it appears that myosin cannot be considered as a genuine centrosomal protein. We have resolved the paradoxical results with CTR56 by showing that in addition to the cellular myosin heavy chain, this antibody also recognizes a high molecular weight protein specifically enriched in centrosomal fractions. The possible biological significance of this finding is discussed in structural and functional terms. © 1992 Wiley-Liss, Inc.
    Additional Material: 6 Ill.
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  • 9
    ISSN: 0886-1544
    Keywords: cytoskeleton ; cell morphogenesis ; immunofluorescence ; antimyosin monoclonal antibody ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A monoclonal antibody, CC212, raised against ciliated cortices of quail oviduct cells and characterized as an antimyosin of smooth muscle and nonmuscle cells, was shown to specifically label a regular cortical network in Paramecium and to recognize two Triton X-100-insoluble polypeptides at 130 and 50 kDa. However, no evidence was obtained that these polypeptides are related to myosin.An immunofluorescence study and ultrastructural immunogold localization allowed us to identify the CC212-decorated material as a component of the outer lattice, a submembrane cytoskeletal network which runs along the top of the ridges visible by scanning electron microscopy and delineates the periphery of each cortical unit. The dynamics of the outer lattice during the cell cycle was studied by immunofluorescence and it was found that the outer lattice growth is achieved without disruption of the preexisting meshes by longitudinal elongation and additon of new transverse partitions. A striking disorganization of the outer lattice was observed in a thermosensitive mutant primarily altered in basal body duplication. These observations suggest possible functions of the outer lattice and demonstrate the interdependence of basal body duplication, surface growth, and outer lattice remodelling.
    Additional Material: 7 Ill.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 8 (1987), S. 238-249 
    ISSN: 0886-1544
    Keywords: human lymphoblastoma cells ; microtubule organizing centers ; isolation centrioles ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A procedure adapted from that described by Mitchison and Kirschner [Nature 312:232-237, 1984] was used to isolate centrosomes from human lymphoid cells. High yields of homogeneous centrosomes (60% of the theoretical total, assuming one centrosome per cell) were obtained. Centrosomes were isolated as pairs of centrioles, plus their associated pericentriolar material. Ultrastructural investigation revealed: 1) a link between both centrioles in a centrosome formed by the gathering in of a unique bundle of thin filaments surrounding each centriole; 2) a stereotypic organization of the pericentriolar material, including a rim of constant width at the proximal end of each centriole and a disc of nine satellite arms organized according to a ninefold symmetry at the distal end and; 3) an axial hub in the lumen of each centriole at the distal end surrounded by some ill-defined material.The total protein content was 2 to 3 × 10-2 pg per isolated centrosome, a figure that suggests that the preparations were close to homogeneity. The protein composition was complex but specific, showing proteins ranging from 180 to 300 kD, one prominent band at 130 kD, and a group of proteins between 50 and 65 kD. Actin was also present in centrosome preparations.Functional studies demonstrated that the isolated centrosomes were competent to nucleate microtubules in vitro from purified tubulin in conditions in which spontaneous assembly could not occur. They were also very effective at inducing cleavage when microinjected into unfertilized Xenopus eggs.
    Additional Material: 9 Ill.
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