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  • 1
    Electronic Resource
    Electronic Resource
    Two different monoclonal antibodies to alpha-tubulin inhibit the bending of reactivated sea urchin spermatozoa (1982)
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 2 (1982), S. 599-614 
    Details
    ISSN: 0886-1544
    Keywords: monoclonal antibodies to tubulin ; radioimmune assay ; immunoautoradiography ; Western blots ; immunofluorescence ; tubulin heterogeneity ; eukaryotic flagellar motility ; immunomotility ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Two monoclonal antibodies reactive for α-tubulin but not for β-tubulin have been prepared, characterized in terms of their relative binding to tubulins from differnt sources by a solid-phase binding assay, immunoautoradiography, and indirect immunofluorescence, and utilized to study flagellar motility. Our results demonstrate that α-tubulins from different species, and even from different tissues of the same species, are nonidentical. Especially interesting was the observation that one of the antibodies, Ab2, immunofluorescently stained microtubules of chick embryo fibroblast cells, but was completely unreactive for microtubules of rat kangaroo (PtK2) fibroblasts; a different antibody, Ab1, stained both cell types. Results of these and additional experiments clearly show that Ab1 and Ab2 recognize discrete and different epitopes on α-tubulin.Monoclonal antitubulins Ab1 and Ab2 each inhibited the bend amplitude of reactivated sea urchin spermatozoa without affecting beat frequencies or the ability of the outer doublet microtubules to slide past each other in elastase-digested models. These results, together with those obtained previously using rabbit polyclonal antitubulin antibodies [Asai and Brokaw, 1980], demonstrate that inhibition of bend amplitude is a common property of antitubulin antibodies and is not due to the binding of antibodies to one specific site on the axoneme. Our results suggest that tubulin subunit conformational changes may occur on the outer doublet lattice and may be integrally involved in the mechanism and control of flagellar bending.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/cm.970020608
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  • 2
    Electronic Resource
    Electronic Resource
    Taxol induces microtubule assembly at low temperature (1981)
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 1 (1981), S. 445-454 
    Details
    ISSN: 0886-1544
    Keywords: taxol ; microtubules ; polymerization ; tubulin ; mitotic inhibitor ; protein self-assembly ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Dissociated bovine brain microtubule protein has been shown to reassemble at 0°C in the presence of the drug taxol. Tubulin polymerization was monitored both by electron microscopy of the polymeric structures and by incorporation of tritiated GTP into filterable polymeric structures. Most of the labeled guanine nucleotide uptake into tubulin polymeric structures occurred in the first 30 minutes of incubation with the drug. The initial polymerization event results in the formation of protofilamentous tubulin ribbons. The first microtubules were noted after 1 hour of incubation with the drug. After 20 hours of incubation at 0°C with taxol, the bulk of the polymerized tubulin appeared to be in the form of microtubules. Cold-stable tubulin rings with a mean diameter of 34 nm were present in the reaction mixture before the addition of taxol and throughout the 20-hour incubation. Most of the rings were apparantly not involved in the taxol-induced microtubule assembly. The results are consistant with a model whereby taxol induces an initial formation of protofilamentous ribbon structures, mostly from free tubulin dimers, and a slower subsequent folding of the ribbon structures into microtubules.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/cm.970010405
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  • 3
    Electronic Resource
    Electronic Resource
    Regulation of cell motility, morphology, and growth by sulfated glycosaminoglycans (1986)
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 6 (1986), S. 273-281 
    Details
    ISSN: 0886-1544
    Keywords: heparin ; glycosaminoglycans ; fibronectin ; cell growth factors ; cell migration ; cell adhesion ; cell morphology ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Due to the recent observation that heparin binds to several growth factors and cell adhesion molecules, the effect of heparin on biological processes governed by growth factors and cell adhesion molecules was investigated. Pharmacological doses of heparin were found to alter cell growth rate, cellular morphology, and cell motility.Concentrations (μg/ml) of heparin or dextran sulfate decreased cell growth rate, but not the final cell density attained in plateau phase. The effect of heparin on cell growth rate was most pronounced when cells were cultured in low concentrations of serum. A heparin-induced decrease in cell growth rate could be reversed by addition of platelet-derived growth factor (PDGF), a heparin-binding growth factor.Heparin altered the morphology of all cell lines studied to various degrees. The effect of heparin on cell morphology was quantitated by measuring the heparin-induced change in cell surface area. HT-1080 and HeLa cells nearly doubled in surface area upon exposure to 10μg/ml heparin. Since several heparin-binding cell adhesion proteins mediate both cell spreading and cell migration, the influence of heparin on cell migration was investigated with an improved version of the phagokinetic track technique. Low concentrations of heparin and dextran sulfate were found to increase the rate of cell migration in a dose-dependent fashion.Since the quantitative effect of heparin on cell growth rate, morphology, and migration depends on the cell line studied, it is suggested that three separate phenomena may be involved. The results presented indicate a central role for sulfated glycosaminoglycans in the control of both cell growth and cell-cell interactions.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/cm.970060304
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  • 4
    Electronic Resource
    Electronic Resource
    Thrombin, epidermal growth factor, and phorbol myristate acetate stimulate tubulin polymerization in quiescent cells: A potential link to mitogenesis (1992)
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 23 (1992), S. 265-278 
    Details
    ISSN: 0886-1544
    Keywords: growth factors ; phorbol 12-myristate 13-acetate ; microtubule-tubulin equilibrium ; initiation of DNA synthesis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Previous studies suggest that alterations in the microtubule (MT)-tubulin equilibrium during G0/G1 affect mitogenesis. To determine the effect of growth factors on the MT-tubulin equilibrium, we developed a radioactive monoclonal antibody binding assay (Ball et al.: J. Cell. Biol. 103:1033-1041, 1986). With this assay, 3H-Ab 1 - 1.1 binding to cytoskeletons in confluent populations of cultured cells is proportional to the number of tubulin subunits polymerized into MTs. We now show that purified α-thrombin increases 3H-Ab 1 - 1.1 binding to cytoskeletons of serum-arrested mouse embryo (ME) fibroblasts from 1.5- to 3-fold. This stimulation is dose-dependent and correlates with concentrations of thrombin required for initiation of DNA synthesis. Other mitogenic factors, epidermal growth factor (EGF) and phorbol 12-myristate 13-acetate (PMA), also stimulate MT polymerization. Addition of colchicine (0.3 μM) eight hours after growth factor addition blocks stimulation of 3H-thymidine incorporation by thrombin, EGF, or PMA, suggesting that tubulin polymerization or subsequent events triggered by MT polymerization are required for cells to enter a proliferative cycle. Consistent with models for autoregulation of tubulin synthesis, thrombin, EGF, and PMA all increase tubulin synthesis 9 to 15 hr after growth factor addition, raising the possibility that the decrease in free tubulin and subsequent stimulation of tubulin synthesis is linked to progression of cells into a proliferative cycle. Colchicine addition to these cells also stimulates DNA synthesis, but colchicine-stimulated cells enter S phase 6 to 8 hr later than those stimulated by growth factors. This delayed stimulation may be related to the time required for degradation of tubulin- colchicine complexes below a critical level. These data suggest that regulation of cell proliferation may be linked to increased MT polymerization and the resulting decrease in free tubulin pools. © 1992 Wiley-Liss, Inc.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/cm.970230406
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  • 5
    Electronic Resource
    Electronic Resource
    Three microtubule-organizing centers are required for ascus growth and sporulation in the fungus Sordaria macrospora (1992)
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 22 (1992), S. 257-273 
    Details
    ISSN: 0886-1544
    Keywords: fungal cytoskeleton ; microtubules ; nocodazole ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The microtubule system of the Sordaria macrospora ascus was examined by antitubulin immunofluorescence, without the removal of the cell wall. The complex cytoskeleton revealed three possible microtubule-organizing centers (MTOCs): the spindle pole body (SPB), the nuclear envelope, and an apical organizing center. MPM-2, a mitotic phosphoprotein antibody which reacts with MTOCs, stained the apical center in a developmentally specific manner, and the nuclear envelope and SPB in a cell cycle-dependent fashion. Nocodazole was used in both high (10-15 μg/ml) and low (0.5 μg/ml) concentrations to depolymerize the networks and reveal their points of origin and recovery. The apical center was active from prophase I to the end of first meiosis. The nuclear envelope was the site of microtubule nucleation in early prophase and at the telophase/interphase transition, while SPBs were active in both nuclear division and sporulation.Mutant strains deficient in sporulation and with aberrant morphology were analyzed by antitubulin and MPM-2 immunofluorescence. Shape mutants showed abnormal or absent apical organizing centers and abnormal cortical microtubule patterns, indicating a possible role for the cortical network in the establishment and maintenance of ascus shape. © 1992 Wiley-Liss, Inc.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/cm.970220406
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  • 6
    Electronic Resource
    Electronic Resource
    The variable fine structure of elastin visualized with verhoeff's iron hematoxylinThis research was supported by N.I.H. Grants AM-10956, AM-11028 and VA TR-168a. (1975)
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 181 (1975), S. 83-94 
    Details
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Verhoeff's iron hematoxylin (VIH) followed by lead citrate (LC) applied to epoxy thin sections stained the dense component of elastic fibers heavily and the peripheral microfibrillar component lightly in guinea pig trachea and mouse testis fixed with a glutaraldehyde-osmium tetroxide sequence. This method stained large fimbriated fibers beneath tracheal epithelium, small fibers and stacked aggregates thereof in the deep lamina propria, cartilage and adventitia of the trachea and large stacked fibers in the fibroelastic band of the trachea. Fibers of the fetus differed from those of the adult, especially in the subepithelial elastic lamina of the trachea. Elastic fibers were intimately associated with fibroblasts and particularly slender fibroblast processes in tracheal stroma and with chondrocytes in tracheal cartilage. Fibroblasts associated with elastic fibers in the tracheal subepithelial lamina propria were often closely bordered by eosinophils, mast cells, or monocytes. Occasional mast cells extended slender processes around elastic fibers in the subepithelial lamina propria. In mouse testis and in many regions of the trachea, small elastic fibers were identified which were below the limits of resolution for the light microscope and were not apparent at the ultrastructural level in routinely stained thin sections.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/ar.1091810107
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  • 7
    Electronic Resource
    Electronic Resource
    Morphogenesis of the truncus arteriosus of the chick embryo heart: Movements of autoradiographic tattoos during septation (1987)
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 218 (1987), S. 434-440 
    Details
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: In order to trace tissue movements during septation of the embryonic truncus arteriosus into aortic and pulmonary cardiac outlets, the cephalic margin of the developing tubular heart of chick embryos was tattooed at Hamilton-Hamburger Stages 20-22 using diffusion micropipettes filled with 0.5% agarose and radioactive macromolecular precursors (tritiated thymidine, uridine, and leucine). Following further incubation for 2, 48, or 96 hours, the locations of such tatoos were determined by autoradiography of sectioned tissue and computer reconstruction of the developing outflow tract.Two hours after tattooing, radiolabeled cells were clustered at the right distal margin of the myocardial tube, as intended. Two days later, during septation of the outflow tract into the two arterial streams, label was concentrated along the posterior margin of the myocardium, between the developing aortic and pulmonary valve anlagen to the embryo's right and left, respectively. Four days following tattooing, as truncal septation neared completion, remaining label was found primarily to the left of the aortic valve ring posterior to the pulmonary outlet. The movements of thymidine tattoos during septation were demonstrated in a series of 31 embryos, 14 fixed at 2 hours, 12 at 2 days, and 5 at 4 days following tattooing; similar results were seen in uridine and leucine labeled hearts.The motion of such tattoos in the developing chick heart suggests that the left side of the definitive semilunar valve ring derives from the right distal margin of the primitive tubular heart and that normal morphogenesis of the great arterial streams involves both retraction and rotation of the embryonic truncus arteriosus.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/ar.1092180411
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  • 8
    Electronic Resource
    Electronic Resource
    A computer graphic study of cardiac truncal septation (1983)
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 206 (1983), S. 207-214 
    Details
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Septation of the cardiac truncus arteriosus in the normal rat embryo was described and illustrated by three-dimensional reconstruction from serial sections through the developing outflow tract. Tracings of selected structures were digitized on an Apple II-plus microcomputer, relayed through a central computer facility, and displayed on a Hewlett-Packard 9845C graphics computer. Rotation and dissection of the color images allowed a kinetic description of the principal features of truncal septation and comparison with analogous events in the chick.A complex of tissue structures appeared in the downstream, distal truncus at 13 days of gestation and moved toward the ventricle(s) through the following day. As in the chick, the forming semilunar valves remained within 200 μm of the cephalic margin of the myocardial sleeve as they rotated and descended toward their definitive position against the heart. Mesenchymal condensations formed within the intervening aorticopulmonary (AP) septum and remained in close association with the principal bifurcation of the spiralling blood lumens, passing between the valves and descending along the conus septum. As the conus septum fused at 15 days of gestation, three outlets for cardiac outflow were seen: the major left and right streams to the aortic and pulmonary valves, respectively, and an apparently dwindling stream which crossed from the right ventricle behind the conus septum to exit through the aorta. In the chick, but not in the rat, nerve bundles invaded the distal truncus during septation to form the cardiac plexus.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/ar.1092060209
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  • 9
    Electronic Resource
    Electronic Resource
    Cytoplasmic stress fibers in the developing heart (1989)
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 223 (1989), S. 82-89 
    Details
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Rhodamine-conjugated phalloidin staining was used to study the distribution of filamentous actin in the developing heart of embryonic chicks and rats during the morphogenetic period of cardiac septation. In the chick, intense fluorescence indicative of abundant filamentous actin was observed along the myocardium and in the mesenchymal condensations that formed within the aorticopulmonary septum at day 5. Such cellular condensations and concentration of filamentous actin were not seen in the atrioventricular cushions nor in the preseptation outflow tract. Similar results were found in the 14-day rat embryo. In electron micrographs, microfilament bundles with irregular dense bodies were seen in elongated mesenchymal cells between the valve sites of both species. Cell-cell contacts were observed between such elongated cells and myocyte processes protruding from the nearby myocardial sheath. These histochemical and ultrastructural observations suggest that such mesenchymal condensations serve a specialized mechanical tensile role during embryonic septation of cardiac outflow channels.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/ar.1092230112
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  • 10
    Electronic Resource
    Electronic Resource
    Phagocytosis of supernumerary spermatozoa by two-cell mouse embryosThis study was supported by a U.S.P.H.S. Research Grant (RO1HD05725) and Research Contract (69-2220). (1974)
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 178 (1974), S. 3-13 
    Details
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Phagocytosis of supernumerary spermatozoa contained within the perivitelline space was observed in 7 of 28 two-cell mouse embryos cultured for 45 hours post-insemination (approximately 20-24 hours after cleavage). The spermatozoa were incarcerated as a result of elevations of the blastomere cytoplasm which gradually surrounded the sperm, overlapped and fused, thus forming a typical phagocytic vacuole. Phagocytosis was not observed in two-cell embryos cultured for less than 20-24 hours after cleavage; this indicates that the blastomeres of two-cell mouse embryos in vitro require approximately 24 hours to develop one of the characteristics of somatic cells, i.e., the ability to recognize and phagocytize foreign material.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/ar.1091780103
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