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  • Cell & Developmental Biology  (13)
  • 1990-1994  (13)
  • 1
    Electronic Resource
    Electronic Resource
    Proliferation-associated oxygen consumption and morphology of tumor cells in monolayer and spheroid culture (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 153 (1992), S. 44-52 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The oxygen consumption rate, proliferative activity, and morphology of EMT6/Ro mouse mammary sarcoma cells in monolayer and multicellular spheroid culture have been investigated in a comparative study. During the transition of monolayer cells from the exponential into the plateau growth phase, there is a distinct decrease in the cellular volume that is associated with a corresponding decrease in the proliferative and respiratory activity of the cells. The decline in cell volume is mainly due to a decrease in the content of cytoplasm, whereas the size of the nucleus is only slightly reduced. A concomitant decrease in the number of mitochondria per cell obviously accounts for the reduction in cellular oxygen uptake. Despite a continuous decrease of cell proliferation from the surface to interior regions of EMT6 spheroids reflected by a gradient in tritiated thymidine labeling, volume-related oxygen consumption is rather uniform in viable regions of these aggregates. The finding can be explained by the results of the morphometric evaluation showing a uniform volume density of mitochondria, i.e., of oxygenconsuming sites within these spheroids. © 1992 Wiley-Liss, Inc.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041530108
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  • 2
    Electronic Resource
    Electronic Resource
    Invadopodia promote proteolysis of a wide variety of extracellular matrix proteins (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 158 (1994), S. 299-308 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Chicken embryo fibroblasts (CEF) transformed by Rous sarcoma virus invade the extracellular matrix (ECM) using plasma membrane protrusions, termed invadopodia, that contact and dissolve the matrix. NormalThroughout this manuscript normal cells refers to chicken embryo fibroblasts and transformed cells refers to these cells infected with Rous sarcoma virus. cells neither form invadopodia nor degrade the ECM. Here we show that cells expressing invadopodia degrade and enter into a fibronectin-rich matrix produced by normal fibroblasts. Within 6 h after seeding onto the matrix, the invasive cells create an area devoid of matrix fibrils surrounding the cell body. Proteolysis mediates this matrix clearing because sevenfold more radiolabeled matrix is released into the growth media by the transformed cells relative to the normal cells. In addition to this assembled matrix, transformed cells were grown on thin layers of purified ECM proteins, revealing that invadopodia can degrade fibronectin, collagen type I, collagen type IV, and laminin. A 160 kDa protease that is extracted from transformed cells by Triton X-114 partitions into the detergent phase and is prominent in ventral plasma membranes that contact the ECM suggesting that it is a membrane associated protease. © 1994 Wiley-Liss, Inc.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041580212
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  • 3
    Electronic Resource
    Electronic Resource
    Epitope mapping and functional studies with three monoclonal antibodies to the C-KIT receptor tyrosine kinase, YB5.B8, 17F11, and SR-1 (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 158 (1994), S. 545-554 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Three monoclonal antibodies (MAbs) to the human c-kit receptor tyrosine kinase (P145c-kit), derived in independent laboratories, have been extensively used in studies of c-kit expression and the role of its ligand, steel factor (SLF), in hemopoiesis and mast cell differentiation and function. In this study, the relationship between the epitopes they identify, and their effects on SLF binding, receptor internalization, and signal transduction are compared.Epitope mapping studies carried out on the high P145c-kit-expressing cell line HEL-DR showed that SR-1 identifies an epitope independent of those bound by YB5.B8 and 17F11, while the latter two antibodies bound to distinct but interacting epitopes. SR-1 potently blocked the binding of SLF to P145c-kit on these cells and also on cells of the factor-dependent line MO7e. In contrast, YB5.B8 and 17F11 had minimal effects on ligand binding. Conversely, SLF partially blocked the binding of SR-1 and YB5.B8 to cells, while binding of 17F11 was actually enhanced by SLF on some target cells. Preincubation of HEL-DR and MO7e cells with MAbs prior to exposure to SLF revealed that 17F11 itself brought about partial down-regulation of P145c-kit and did not inhibit SLF-mediated down-regulation. SR-1 caused minimal down-regulation and inhibited SLF-mediated receptor internalization. YB5.B8 had minimal effects on either cell line in this assay. To determine whether the antibodies had any agonist activity, they were compared with SLF for their ability to bring about receptor phosphorylation in intact MO7e cells. All three antibodies induced detectable tyrosine phosphorylation with 17F11 being the most effective, while YB5.B8 was the least effective. Finally, the ability of the antibodies to influence the proliferation of the MO7e cells was examined. As expected, SR-1 potently inhibited the proliferative response to SLF, while 17F11 weakly inhibited and YB5.B8 had negligible effect. In the absence of SLF both 17F11 and YB5.B8 displayed very weak but reproducible agonist activity. © 1994 Wiley-Liss, Inc.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041580321
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  • 4
    Electronic Resource
    Electronic Resource
    Immunoelectron microscope localization of snRNP, hnRNP, and ribosomal proteins in mouse spermatogenesis (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 35 (1993), S. 261-271 
    Details
    ISSN: 1040-452X
    Keywords: Immunocytochemistry ; Ultrastructure ; Perichromatin granules ; Interchromatin granules ; Mouse spermatids ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have studied the ultrastructural distribution of heterogeneous nuclear ribonucleoproteins (hnRNPs), small nuclear ribonucleoproteins (snRNPs), and ribosomal proteins during mouse spermatogenesis and spermiogenesis by means of specific antibodies and immunocytochemistry.All the above components were detectable from primary spermatocytes until the spermatid elongation phase, when the RNA synthetic activity is known to cease. Ribosomal protein (P1/P2 and L7) labeling disappeared as early as during the acrosome phase, and nucleoli were no longer labeled even during the cap phase. The nucleoplasmic structures labeled with the different anti-nucleoplasmic RNP immunoprobes corresponded, until the acrosome phase, to those previously observed as targets of the same antibodies in the nucleoplasm of somatic cell nuclei. Clusters of interchromatin granules of spermatocyte and early spermatid nuclei exhibit some labeling for hnRNP when compared with nuclei of Sertoli cells or previously analyzed liver or tissue culture cells, where these structural constituents usually remain weakly labeled or unlabeled.In spermatids in step 10, another type of nuclear granule, resembling perichromatin granules, but occurring in aggregates, can be observed. These structural constituents were labeled with antibodies recognizing nucleoplasmic snRNP antigens and therefore suggesting a non-nucleolar origin of these granules.Finally, we have observed nucleoplasmic areas of fibrogranular material, occurring only in primary spermatocytes. These components were labeled with anti-ribosomal protein antibodies but did not contain either hnRNPs or snRNPs. © 1993 Wiley-Liss, Inc.
    Additional Material: 15 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/mrd.1080350308
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  • 5
    Electronic Resource
    Electronic Resource
    Quantitative aspects of synapses on Golgi-impregnated neurons (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 23 (1992), S. 334-352 
    Details
    ISSN: 1059-910X
    Keywords: Identified neurons ; Quantification ; Rotating/tilting ; Synaptic contacts ; Electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: With the classical Golgi techniques, numerous types of neurons can be distinguished in the cerebral cortex, each with a specific dendritic geometry and pattern of axonal ramifications. In the present review we describe two techniques which allow quantification of synapses on identified neurons: (1) Golgi-rapid impregnation-gold toning-electron microscopy, and (2) Golgi-Kopsch impregnation-gold toning-electron microscopy in combination with staining of the tissue with ethanolic phosphotungstic acid (E-PTA). Both techniques were applied on neurons in the visual cortex of young and adult rabbits. By means of rotating and tilting specimens in the electron microscope, the nondistinctive ultrastructure of obliquely sectioned synapses can be circumvented, leading to precise estimates of asymmetrical vs. symmetrical synapses without complete reconstruction of the neuron. © 1992 Wiley-Liss, Inc.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070230408
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  • 6
    Electronic Resource
    Electronic Resource
    In vitro maturation of the potential for movement of carp spermatozoa (1991)
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 29 (1991), S. 259-270 
    Details
    ISSN: 1040-452X
    Keywords: Sperm motility ; Fish ; Motility ; Sperm ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Carp semen obtained from isolated fish after hormonal stimulation was highly variable in terms of volume of semen, osmotic pressure of the seminal plasma, and sperm capacity to move. Moreover, this last parameter was unstable when the spermatozoa were kept within the seminal plasma, and the present work was designed to investigate and possibly correct this phenomenon. Sperm potential movement was the major parameter studied and was measured by the percentage of motile cells in a final 3,000-fold dilution in a medium of low osmotic pressure in which sperm movement is known to occur (Morisawa and Suzuki, Science 210:1145-1147, 1980). This was completed with occasional measurements of flagellar beat frequencies and demembranation-reactivation of axonemal movement. The results showed that sperm potential movement was preserved upon dilution of the semen into cold 200 mM KCl medium and that semen of initially “poor” quality or spermatozoa that had lost their capacity to move during storage in the semen recovered gradually their potential movement during incubation at 2°C in the same medium. The K+ dependence for both the conservation and the regeneration of sperm capacity to move showed a minimal requirement of 50 mM KCl in media of high osmotic pressure. Na+ ions had similar properties but not divalent cations. The K+ activation was not pH dependent between pH 9.03 and 6.04. Whatever the functional state of live spermatozoa, demembranation-reactivation occurred in ATP-Mg2+. It is concluded that, with dilution of the semen in appropriate conditions, carp spermatozoa retain or acquire potential movement and therefore are a lower vertebrate spermatozoa model available year-round. In addition, obtaining potentially nonmotile sperm and reversion in vitro might be useful to study the control of in vitro maturation.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/mrd.1080290308
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  • 7
    Electronic Resource
    Electronic Resource
    Transgenic offspring by transcaryotic implantation of transgenic ovaries into normal mice (1990)
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 25 (1990), S. 42-44 
    Details
    ISSN: 1040-452X
    Keywords: Ovary transfer ; Mice ; Transgenic offspring ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Female transgenic mice may be unable to reproduce successfully if the product encoded by the transgene results in pathological changes or affects the fertility of the mouse. To approach this problem, we have produced chimaeras by transferring the ovaries of transgenic mice into normal mice of the same strain. Such chimaeras will be an ideal tool for investigating the interactions between transgenic ovaries and normal mice or vice versa. Here we show that, using this method, we were able to get large numbers of transgenic offspring even from founder transgenic female mice that were themselves infertile as a result of the overexpression of growth hormone genes. Although none of the ovary recipients were given immunosuppressant treatment, 60% of the recipients had biologically active ovaries over a mean period of about 100 days.
    Additional Material: 2 Tab.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/mrd.1080250108
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  • 8
    Electronic Resource
    Electronic Resource
    Ontogeny of the limb skeleton in Alligator mississippiensis: Developmental invariance and change in the evolution of archosaur limbs (1990)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 203 (1990), S. 151-164 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Crocodilians and birds are the extant representatives of a monophyletic taxon known as archosaurs. Their limbs are highly derived in terms of reduction in number of skeletal elements in both the carpus and the tarsus. It is necessary to have a detailed description of crocodilian limb ontogeny to address the evolutionary issue dealing with the origin and organization of the avian limb. In this paper, we present an analysis of the early development of the crocodilian limb skeleton. Contrasting with earlier observations, we redefine the number and composition of carpal, tarsal, and phalangeal elements. This ontogenetic information is then used to introduce a revision of the homologies of the skeletal elements in the crocodilian limb. Some invariances are pointed out in the developmental organization of tetrapod limbs and this evidence serves to readdress several issues concerning the evolution of the avian limb. We present further embryological data in support of the hypothesis that digits 2-3-4 are the components of the wing skeleton in birds. In general, our comparative survey indicates that the elements that appear late in ontogeny are the ones lost in phylogeny. By comparing turtle (primitive) limb development with crocodilian and bird development, we propose a hypothesis in which the derived skeletal patterns found in crocodilians and birds have originated by a heterochronic process of paedomorphosis.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jmor.1052030204
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  • 9
    Electronic Resource
    Electronic Resource
    Natural and experimental reduction of the avian fibula: Developmental thresholds and evolutionary constraint (1992)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 214 (1992), S. 269-285 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Fibula reduction is a key feature of avian limb evolution. In a combined comparative and experimental approach the present study analyses the trends of fibula reduction in extant birds and their developmental basis. The study of 55 species of birds reveals four different types of tibiotarsus-to-fibula relationships. Extremely small fibulae are associated with two types of limb modification: (1) elongations of the limb primarily affect the tibiotarsus, increasing its length more than that of the fibula; (2) miniaturizations of the limb reduce both tibiotarsus and fibula length, but are reglarly associated with structural reductions of the distal parts of the fibula. True structrual reductions are distinguished from relative size reductions. The specific features of fibula reduction are analyzed through experimental mesenchyme excisions in chick limb buds. The methodical variation of experimental parameters resolves a long-standing controversy about the effects of mesenchyme reductions on the patterns of skeletal formation. Mesenchyme excisions are shown to have unequal effects on the two zeugopod bones, affecting the fibula to a greater degree than the tibiotarsus. Several of the features seen in birds with advanced fibula reductions are paralleled by the effects of mesenchyme reductions. The consequences of this differential susceptibility of the skeletal blastemata are discussed both in terms of pattern formation in limb development and in terms of its bearing on the patterns of evolutionary limb reduction. It is concluded that thresholds of cell number and blastema size in development constrain the patterns of phenotypic variation in avian limbs. © 1992 Wiley-Liss, Inc.
    Additional Material: 12 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jmor.1052140304
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  • 10
    Electronic Resource
    Electronic Resource
    Evidence for involvement of a nuclear envelope-associated RNA helicase activity in nucleocytoplasmic RNA transport (1990)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 145 (1990), S. 136-146 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: It seems well established that translocation of at least some mRNAs through the nuclear pore is (1) an energy-dependent process, and (2) dependent on the presence of the poly(A) segment attached to most mRNA species. We describe that RNA helicase (RNA duplex unwindase) activity is present in a nuclear envelope (NE) preparation, which also appears to be involved in nucleocytoplasmic RNA transport. This activity unwinds RNA : RNA hybrids. The helicase has a pH optimum of 7.5 and a temperature optimum of 30°C. Applying the sealed NE vesicle system, it was shown that duplex RNA species are readily released from the vesicles in an unidirectional manner, in contrast to single-stranded RNA, which is much slower transported into the extravesicular space. Attachment of a poly(A) segment to the RNA duplex additionally increases the efflux rate of this RNA. Efflux of duplex RNA but not efflux of single-stranded RNA was strongly inhibited by formycin B 5′-triphosphate. Our results suggest that, besides poly(A), duplex structures, if present in a given RNA, modulate and control the export of RNA.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041450119
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