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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Morphology 190 (1986), S. 191-200 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Species of the salamander genus Plethodon have a characteristically uniform morphology. Morphological conservatism at the level of interspecific comparisons, however, is not always reflected within species. Perhaps the most extreme example of intraspecific variation is the recent description of extensive variability in limb-skeletal patterning both within and between populations of the widespread species P. cinereus. We utilized limb regeneration following experimental amputation as a tool (1) to examine whether naturally occurring variant skeletal patterns result from limb loss and regeneration in nature, and (2) to assay the intrinsic (i.e., genetic) component of between-individual variation in mesopodial patterning. We observed the following. First, regenerate patterns are strikingly different from native patterns: interelement fusions in regenerates are typically between proximodistally adjacent cartilages, whereas interelement fusions in native variant limbs occur exclusively between laterally adjacent cartilages. Fusions also are over ten times more frequent in regenerates than in native limbs. Second, there is no strong correlation between native limb pattern (typical vs. variant) and the regenerate pattern. We conclude that variability in field-collected P. cinereus reflects extensive intrapopulation variation in limb-skeletal patterning during original limb development, rather than regeneration in nature, and that limb regeneration analysis provides no evidence of a strong genetic component to between-individual variation. Finally, unusual mesopodial patterns produced during limb regeneration may be related to the mechanical factors impinging on the regenerating limb in this terrestrial species.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 125 (1985), S. 367-378 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We have reexamined the possible role of the Na+/H+ antiport in the cellular response by PC12 pheochromaytoma cells to nerve growth factor (NGF). In contrast to previous reports, we observe no activation of Na+/H+ exchange in these cells, using a very sensitive assay based on the measurement of cytoplasmic pH with dimethylfluorescein dextran (Rothenberg et al., J. Biol Chem., 258: 4883-4809, 1983). Our measurements indicate that the PC12 pheochromacytoma cells, under all conditions tested, show a high rate of Na+/H+ exchange. The discrepancy between these observations and previous experiments could be due to differences in cells in different laboratories, but also to changes in cell adhesion induced by NGF. We describe conditions where intracellular pH and rates of Na+ uptake can be measured reliably in PC12 cells with adequate controls for cell adhesion. We conclude that activation of Na+/H+ exchange is neither sufficient nor required for the differentiation of PC12 cells induced by NGF.
    Additional Material: 8 Ill.
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 124 (1985), S. 131-136 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The Ca++ requirement for in vitro lymphocyte stimulation by lectins is well known and can be demonstrated by the use of Ca++ chelators. In this study, three Ca++ antagonists were examined for their effects on lymphocyte proliferation. [3H]-thymidine incorporation was employed to measure DNA synthesis in several systems. Stimulation and proliferation were achieved by the addition of one of the following: the mitogenic lectin concanavalin A (ConA); the combination of two co-mitogens, the calcium ionophore A23187 and the phorbol ester, 12-0-tetradecanoylphorbol-13-acetate (TPA), neither of which is mitogenic alone; or the non-mitogenic lectin, wheat germ agglutinin (WGA) with TPA. These mitogenic systems were tested for their sensitivity to the Ca++ channel blockers verapamil and nicardipine and the intracellular Ca++ antagonist TMB-8. We found that the ConA and WGA plus TPA treated cells were inhibited approximately 50% by 10 μM verapamil, nicardipine or TMB-8. The stimulation caused by A23187 and TPA was only inhibited by TMB-8 and nicardipine. The inhibitory effects caused by the Ca++ antagonists could not be reversed by the addition of exogenous Ca++ (0.1-1.5 mM), but were reversed by repeated washings in antagonist free media. Using TMB-8 we saw an apparent intracellular Ca++ dependence throughout the G1 phase. Previous studies using Ca++ chelators or Ca++ antagonists suggested an endpoint at about halfway through this period.
    Additional Material: 3 Ill.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 141 (1989), S. 301-309 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The effect of removal of PC12 cell nuclei on neurite outgrowth was studied. Enucleation (80-90%) was accomplished in the presence of cytochalasin B using a centrifugation technique that exploited the very tight adhesivity of PC12 cells for a substratum composed of an extracellular matrix secreted by bovine corneal endothelial cells in response to epidermal growth factor treatment. Neither nucleated nor enucleated PC 12 cells showed significant neurite outgrowth on this particular matrix in the absence of nerve growth factor. In the presence of nerve growth factor both PC12 cell types initiated neurite outgrowth, but whereas neurites from nucleated cells grew continuously for two days, those from enucleated cells reached a maximum length after one day. The results suggest that neurite initiation but not continued neurite growth or stabilization can occur in the absence of transcription.
    Additional Material: 4 Ill.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 122 (1985), S. 171-177 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A primary cloning technique has been employed for the isolation of nine spontaneously transformed cell lines from mouse skeletal muscle. Four of these lines were isolated after selection for partial resistance to the purine (adenine) analog 2′6′diaminopurine and five were isolated from non-selected control dishes. Four of the nonselected lines and three of the selected lines demonstrated a fibroblastoid morphology in vitro. The other two cell lines (one from each group) were epithelioid. Two of the three selected fibroblastoid lines were found to contain significant quantities of the enzyme 5′nucleotidase (EC3.1.3.5), whereas the four nonselected fibroblastlike lines, one selected fibroblastlike line, and the two epithelioid lines did not. In the two cell lines expressing 5′nucleotidase activity, this expression was stable in the absence of selective pressure. Histochemical staining of mouse skeletal muscle for 5′nucleotidase activity demonstrated positive staining in the cells of small blood vessels and in a subset of the connective tissue cells. The bulk of the skeletal muscle tissue, however, had no detectable 5′nucleotidase activity. We propose that the two cultivatable types of fibroblastoid cell lines represent distinct classes of fibroblastlike cells in vivo, reflecting alternative states of stable cellular differentiation involving 5′nucleotidase expression.
    Additional Material: 3 Ill.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 125 (1985), S. 192-199 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: rac-1-O-Myristoyl-2-O-acetylglycerol, rac-1-O-palmitoyl-2-O-acetylglycerol, and rac-1-O-oleoyl-2-O-acetylglycerol acted like phorbol myristate acetate and mezerein in stimulating human neutrophil aggregation. Responses to these agents were equally influenced by cytochalasin B, extracellular calcium and magnesium, arachidonate antimetabolites, and procedures that rendered the cells desensitized to other agonists. The compounds also inhibited the binding of [3H]-phorobol myristate acetate to its receptor on neutrophils. Thus, these agents are biologically homologous. They act by binding to a common receptor. This receptor may function physiologically as a transducer for endogenous glycerides that form in cells challenged by other stimuli.
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