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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 21 (1992), S. 101-110 
    ISSN: 0886-1544
    Keywords: F-actin ; silk gland ; phalloin ; periluminal circumferential actin bundles ; actin-coated vacuoles ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Labeling of silk glands with rhodaminyl-phalloin shows that most F-actin is restricted to parallel bundles that form rings around the gland lumen at the apical cell surface. The bundles are lost when larval feeding stops at moulting, and the F-actin is redistributed through the cytoplasm as coats to vacuoles and, occasionally, in variably oriented strands. After moulting there is a return to the distribution of filamentous actin in the apical periluminal rings of bundles. These events occur at the same time as F-actin in the nuclear shell [Henderson and Locke, submitted] undergoes its own set of changes. In silk gland cells two kinds of f-actin deployment take place concurrently.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 23 (1992), S. 169-187 
    ISSN: 0886-1544
    Keywords: nuclear actin ; nuclear myosin ; nuclear shell ; nuclear shape ; nuclear matrix ; silk gland ; nuclear structure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The branched nuclei from silk gland cells of larvae of Calpodes ethlius label with antibodies to actin and myosin and with rhodaminyl-phalloin, which is specific for f-actin. Optical sectioning localizes this actin and myosin to the nuclear periphery. Residual nuclear-associated fractions prepared from these cells contain sheets of nuclear lamina-like structures that bind heavy meromyosin and gold-tagged antibodies to actin and myosin. The results suggest that both actin and myosin, or a myosin-like protein, are components of a layer at the nucleocytoplasmic boundary that we call the nuclear shell. The nuclear shell appears to be associated with the nuclear envelope and may correspond to a zone on the cytoplasmic face of the envelope seen in electron micrographs of unextracted cells. The residual nuclear-associated fraction has a unique isoform of actin (43 kD, pl 6.45) that might allow the nuclei to associate with an actin network structurally and developmentally distinct from that of the cytoplasm. © 1992 Wiley-Liss, Inc.
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 3 (1983), S. 683-691 
    ISSN: 0886-1544
    Keywords: α-spectrin ; coelomocytes ; filopodia ; actin/membrane interactions ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We have investigated the presence and localization of an α-spectrinlike protein and its potential role in the morphological transformation of sea urchin coelomocytes. In immunofluorescence images there is a diffuse fluorescence throughout the petaloid cytoplasm, indicating a random distribution of the spectrinlike protein prior to the transformation. As these cells form filopodia, there is a coincident appearance of a spectrinlike protein, as seen in fluorescent images, at the site of filopodial initiation. As the filopodia continue to form and lengthen, the spectrin localization parallels their development. There is a single polypeptide observed on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels of whole coelomocyte lysates that cross-reacts with the anti-α-spectrin immunogen and comigrates with it at 240 kilodaltons.
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  • 4
    ISSN: 0886-1544
    Keywords: microtubule bending ; cytoskeletal assembly ; cochlea ; mouse ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Mature inner pillar cells in the mammalian organ of Corti are curved through about 60°, where they arch over adjacent epithelial cells and the apex of an intercellular space called the tunnel of Corti. This report deals with changes in microtubule organization that are associated with cell bending and tunnel formation during morphogenesis of the mouse organ of Corti.A large bundle of up to 3,000 microtubules assembles in each inner pillar cell. Microtubule rearrangement occurs about 5 days after bundle assembly begins. The lumen of each initially straight hollow tube-shaped microtubule bundle is occluded as the bundle becomes more compact and elliptical in cross section. This event anticipates the once-only bending which subsequently occurs between particular levels (abut 9-19 μm) below the top of a bundle as it curves into its final shape about 2 days later. Microtubule rearrangement presumably facilitates bending which is effected in the plane of lest mechanical resistance parallel to the short axis of a bundle's elliptical cross-sectional profile.Precocious bending of bundles has been induced about 1.5 days in advance of the natural event. Abnormal positioning of these prematurely curved bundles indicates that bending is effected by a contractile mechanism located within bundles rather than being a response to externally applied forces. The potential importance of such microtubule-associated contractions for active modulation of the vibratory response in the cochlea during hearing is considered. © 1993 Wiley-Liss, Inc.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 28 (1994), S. 165-178 
    ISSN: 0886-1544
    Keywords: WISH ; Keratin ; 3-D reconstruction ; mitosis ; intermediate filaments ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Three dimensional (3-D) reconstruction of four mitotic WISH cells from ultrathin sections gave an informative representation of the spatial distribution of keratin densities in these cells. The correspondence between the densities as studied by transmission electron microscopy (TEM) and the Keratin bodies initially revealed by immunoflourescent colabeling of cultures, was confirmed by immunoelectron-microscopy. The smaller, and sometimes more elongated densities, were relatively abundant just beneath the subplasmalemmal microfilament band; and at certain levels of the mitotic cell they were observed to be connected to neighboring densities by intact intermediate filaments (IFs). The larger and more spherical densities appeared to be somewhat more discrete and randomly distributed. Other observed associations of the keratin densities included the telophase contractile ring of microfilaments, chromosomes, the reformed telophase nucleus, and desmosomal junctions with neighboring interphase cells. Cytochalasin D (CD) treatment of cells displaced the peripheral keratin densities toward the cell membrane. The density volume constituted 0.52% to 1.57% of the total cell volume, and the proportional density size was decreased in the cells that had progressed into anaphase and telophase. The observed formation and subsequent dissolution of keratin densities during mitosis may represent a dynamic mechanism of restructuring the keratin cytoskeleton in an unpolymerized form in order to allow for rapid reformation of interphase cell junctions. The physical associations observed between intact IFs and the keratin densities may provide support at certain depths of the mitotic cell, and the juxtaposition of densities with nuclear components suggests a possible source of and role for keratin IFs during nuclear events. © 1994 Wiley-Liss, Inc.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 154 (1993), S. 229-237 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Chronic anoxia, glucose starvation, low pH, and numerous other conditions induce the glucose-regulated system of stress proteins (GRPs), whose principal members are observed at 78, 94, and 170 kDa. These stresses may be expected to occur during growth in untreated tumors. To examine the possibility that GRPs are correspondingly induced, we have examined the protein profiles of small (〈0.1 g), intermediate (0.2-0.8 g), and large (〉1.8 g) radiation-induced fibrosarcoma (RIF) tumors grown on C3H mice. One and two-dimensional gel electrophoresis indicate that the principal GRPs at 78 and 94 are coordinately and substantially increased in large tumor masses, relative to the small, and may be partially increased in the intermediate tumors. Necrotic material removed from large tumors exhibited an identical pattern of GRP induction with no visible indication of protein degradation and also contained a significant fraction of viable cells. Western blot analysis using rabbit antisera raised against the 78 and 170 kDa GRPs also demonstrated the enhanced accumulation of these proteins in the large tumors. The antibody against the 170 kDa GRP was also capable of detecting the induction of this stress protein in large tumors by indirect immunofluorescence analysis. Northern blot studies using a probe for the GRP 78 gene also showed an increase in GRP 78 message in large tumors as well as in RIF cells exposed to anoxic stress in vitro. Two-dimensional gel electrophoresis indicated that the major heat shock proteins at 70 and 90 kDa were not increased in the larger tumors, and the amount of the 90 kDa species was reduced. Finally, the quantity of vimentin and its degradation products is significantly diminished in large tumors and in anoxic cells. This study demonstrates that RIF tumor cells undergo a glucose regulated stress response in situ during tumor growth. © 1993 Wiley-Liss, Inc.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 16 (1987), S. 323-341 
    ISSN: 0148-7280
    Keywords: zona pellucida ; deglycosylation ; antisera ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Zonae pellucidae (ZP) were isolated from 1,500 porcine ovaries and heat solubilized to generate approximately 15 mg ZP glycoprotein. Analysis of this material by isoelectric focusing, one-dimensional electrophoresis, and gas chromatography indicated the presence of a major glycoprotein species that exhibited considerable microheterogeneity with respect to its charge (pI 7.5-3.5) and molecular mass (45-85 kDa) and that contained 39.6% carbohydrate, predominantly N-acetylglucosamine.Chemical deglycosylation of porcine ZP using trifluoromethanesulphonic acid (TFMS) resulted in the production of five discrete protein bands on one-dimensional sodium dodecyl sulphate/polyacrylamide gel electrophoresis (SDS/PAGE) with molecular masses of 66, 52, 36, 32, and 16 kDa.Antisera raised in rabbits and marmosets to ZP and/or deglycosylated ZP (DGZP) were used in immunoblotting experiments to demonstrate the retention of immunogenicity by DGZP and the cross-reactivity of the antisera with their heterologous antigen. These studies indicated that antisera that were capable of inhibiting the fertility of primates in vivo and the penetration of the human ZP in vitro reacted preferentially with 3 of the 5 products of deglycosylation, with molecular masses of 66, 52, and 36 kDa. Anti-DGZP antibodies were also shown to interact with intact porcine and human ZP and, with the latter, to block the ability of human spermatozoa to both bind to and penetrate this structure.
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  • 8
    ISSN: 0148-7280
    Keywords: deglycosylated polypeptides ; porcine zonae pellucidae ; mammalian oocyte ; contraceptive potential ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The major deglycosylated polypeptides of the porcine zona pellucida (ZP), with molecular masses of 66, 52, 36, and 32 kDa, were purified to homogenity with one-dimensional sodium dodecyl sulphate/polyacrylamide gel electrophoresis (SDS/PAGE). Immunofluorescence studies demonstrated that antibodies to the DGZP fraction, and the 66- and 32-kDa polypeptides, bound predominantly to the outer ZP; however, only the first two of these antisera formed an immunoprecipitate around the outer human ZP. In immunoblotting experiments using polyclonal antisera raised to these molecules all four polypeptides exhibited cross-reactivity with each other and their parental glycoprotein families (ZP 1-4). In addition, the antisera were tested in an in vitro human gamete bioassay to determine their contraceptive potential; antibodies to the 32-kDa deglycosylated polypeptide inhibited human gamete interaction to the greatest extent, 5.3% (± 1.2%), relative to a control value of 100%.
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  • 9
    ISSN: 0197-8462
    Keywords: c-myc ; histone H2B ; dot-blot hybridization ; transcription ; human cells ; Life and Medical Sciences ; Occupational Health and Environmental Toxicology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Physics
    Notes: Human HL-60 cells were exposed for 20 min to an electromagnetic field at frequencies ranging from 15 to 150 Hz and at densities from 0.2 to 2.3 mT (2 to 23 gauss). Following each exposure, quantitative levels of c-myc and histone H2B transcripts were determined by dot-blot hybridization analyses and compared with unexposed control samples. The most pronounced increase in each transcript occurred after exposure at 45 Hz, with levels more than four times that found in unexposed controls.
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  • 10
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 473-483 
    ISSN: 0192-253X
    Keywords: N-glycosylation ; glycoprotein sorting ; oligosaccharide processing ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have used metabolic radiolabeling with oligosaccharide precursors, coupled with subcellular fractionation, to examine the distribution of several classes of asparagine-linked oligosaccharides during early development. In Dictyostelium, we have observed endoglycosidase H (endo H)-sensitive structures with sizes corresponding to 10 (Hex10) and 11 (Hex11) hexose residues on the chitobiose core. Only Hex11 was detected as the major structure on fucosylated endo H-resistant species. All Hex11 species cofractionated with plasma membrane and secreted glycoproteins, whereas Hex10 appeared to be confined to intracellular membrane and soluble glycoproteins. Sulfated species correlated with lysosomal and secreted fractions, and glucose residues were markedly depressed in Hex11 of secreted glycoproteins. Outer branch structural studies have revealed several components of the endo H-sensitive species. Usingα-mannosidase and β-hexosaminidase as diagnostic tools, species elucidated thus far are: a structure with 10 mannoses, a structure with nine mannoses and an intersecting N-acetylglucosamine, structures with three glucoses and seven or eight mannoses and several larger species with multiple blocks to digestion.
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