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  • Wiley-Blackwell  (2)
  • American Chemical Society
  • American Society of Hematology
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  • National Academy of Sciences
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  • Wiley-Blackwell  (2)
  • American Chemical Society
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  • Institute of Physics
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  • 1
    Digitale Medien
    Digitale Medien
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 3 (1983), S. 199-210 
    ISSN: 0886-1544
    Schlagwort(e): sperm ; flagellum ; motility ; cAMP ; freeze-thawing ; Life and Medical Sciences ; Cell & Developmental Biology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie , Medizin
    Notizen: Demembranated and membrane disrupted bull sperm models exhibit an increase in motility when exposed to cAMP. Tritium-labeled cAMP was used to locate the initial site of action of cAMP in the modeled sperm preparations. cAMP did not bind selectively to the modeled cells, and the presence or absence of plasma membrane fragments on the models did not significantly alter this result. When suspension medium taken from modeled sperm preparations was subjected to gel filtration on Sephadex G25-150 columns, cAMP bound to a high molecular weight component that eluted with the void volume. The responsible binding factor is a soluble component that is released when the plasma membranes of the sperm are disrupted during the modeling procedure. To test the importance of the cAMP binding factor, modeled sperm were centrifuged, the super-natant solution was decanted, and the cells were resuspended in fresh medium. After this treat-ment the cells could be restored to motility with Mg-ATP but no longer exhibited a response to cAMP. Furthermore, addition of cAMP binding factor isolated by gel filtration partially restored the response of these sperm to cAMP. Investigation of the properties of the cAMP-binding factor have confirmed that it is specific for cAMP, with a much lower affinity for AMP and cGMP. In the pre-sence of a large excess of unlabeled cAMP the labeled complex has a half-life of approximately 1 hour. Our results indicate that the action of cAMP on the motility of modeled sperm is mediated by its attachment to a high molecular weight, soluble component of the cell cytoplasm.
    Zusätzliches Material: 2 Ill.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 2
    Digitale Medien
    Digitale Medien
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 26 (1993), S. 66-76 
    ISSN: 0886-1544
    Schlagwort(e): nickel ; cadmium ; dynein ; motility ; flagella ; Life and Medical Sciences ; Cell & Developmental Biology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie , Medizin
    Notizen: Bovine sperm, extracted with 0.1% Triton X-100, frozen at -20°C for 48-120 hours, and thawed, disintegrated by microtubule sliding when 1 mM MgATP was added. Microtubules and outer dense fibers (ODFs) were usually extruded in groups or “bundles.” A total of 44.5% of the cells extruded two distinct bundles, one from each side of the connecting piece, exhibiting opposite curvatures. Only one bundle was observed in 46.2% of the cells, and 9.2% showed no signs of sliding. Transmission electron microscopy (T.E.M.) showed one group consisting of the 4,5-6,7 elements, with the 9,1,2 elements on the other side of the axoneme making up the other bundle. T.E.M. revealed that when only one side of the axoneme had extruded elements, they were always from the 4,5-6,7 group. The remainder of the axoneme (8,9,1,2,3 and the central pair) was left relatively intact, suggesting a difference in the sliding response of the nine pairs of axonemal microtubules. These results indicate a predisposition for sliding between elements 7 and 8 over that between doublets 2 and 3, perhaps due to a disparity in activation thresholds. Also, both Ni2+ and Cd2+ appear to selectively block activation of 2-3 interdoublet sliding.Incubation with 0.25 mM Ni2+ prior to adding MgATP modified the percentages of sliding patterns: 8.6% demonstrated two-sided extrusion, 58.2% showed one-sided, and 33.2% had no extruded bundles. Again, when half the axoneme was missing, it was always the 4,5-6,7 group. Ten micromolar Cd2+ altered the sliding pattern similarly to Ni2+, with 28% two-sided extrusion, 55.9% one-sided extrusion and 16.1% with no extruded bundles.Either pretreatment regimen impeded extrusion of the 9,1,2 group in a high percentage of cells, compared to untreated cells. This specific inhibition of the 9,1,2 side by Ni2+ or Cd2+ is especially significant since Ni2+ also inhibits spontaneous wave initiation in bull sperm (Lindemann et al.: Journal of Cell Biology 87:420-426, 1980), and both Ni2+ and Cd2+ reportedly block the flagellar Ca2+-response in rat sperm (Lindemann and Goltz: Cell Motility and the Cytoskeleton 10:420-431, 1988; Lindemann et al.: Cell Motility and the Cytoskeleton 20:316-324, 1991). © 1993 Wiley-Liss, Inc.
    Zusätzliches Material: 6 Ill.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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