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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 161 (1994), S. 435-440 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The goal of this study was to relate insulin-like growth factor (IGF) binding with IGF-stimulated growth in a murine mammary epithelial (COMMA-D/MME) cell line. Affinity crosslinking with [125I]IGF-I showed major bands at 224,000 and 148,000 Mr that were ablated by the inclusion of IGF-I or -II at 130 nM. Scatchard-transformed [125I]IGF-I binding data was best fit by a two-site model, with 24,000 sites possessing a Kd of 0.33 nM and 1,000 sites possessing a Kd of 8.09 nM per cell. Competition analysis showed ED50 values for IGF-I, -II, and insulin to be 0.90 ± 0.03, 7.15 ± 4.27, and 335 ± 104 nM, respectively. Affinity crosslinking of [125I]IGF-II showed three major bands of 230,000, 148,000, and 61,000 to 58,000 Mr. Unlabeled IGF-II ablated all bands, while IGF-I and insulin ablated only the 148,000 Mr band. Competition analysis showed ED50 values for unlabeled IGF-I and -II to be 0.10 ± 0.01 and 5.31 ± 2.04 nM, respectively. In spite of the receptor affinity differences, no significant difference was noted in IGF-I and -II in capacity to stimulate cell growth. These data indicate that COMMA-D/MME cells express IGF receptors and that both IGFs are mitogenic. © 1994 Wiley-Liss, Inc.
    Additional Material: 5 Ill.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 25 (1990), S. 328-338 
    ISSN: 1040-452X
    Keywords: Nuclear run-on ; mRNA half-life ; Strongylocentrotus purpuratus ; Lytechinus pictus ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: mRNAs for Spec 1 and Spec 2 of Strongylocentrotus purpuratus and LpS1 of Lytechinius pictus accumulate only in the aboral ectoderm of developing embryos. In vitro nuclear transcription assays were done to study the transcriptional regulation of these cell type-specific genes. Spec 1, Spec 2c, and Spec 2d genes all appeared to be transcriptionally activated at the late cleavage-early blastula stage of S. purpuratus. Differences in the relative transcription rates during development appeared to play a major role in determining the relative levels of the various Spec mRNAs. The L. pictus LpS1 gene was transcriptionally activated at a similar developmental time as the corresponding S. purpuratus genes. Nuclei from gastrula or pluteus ectodermal and endodermal/mesodermal cell fractions were used to demonstrate that Spec 1 and LpS1 genes were transcriptionally active in ectoderm nuclei but not in endoderm/mesoderm nuclei, suggesting that in vivo the Spec 1 and LpS1 genes are spatially controllod at the transcriptional level. Estimations of the absolute rate constants for Spec 1 transcription were mado at the late cleavage, mesenchyme blastula, and midgastrula stages. Calculations using these rate constants and the known levels of Spec 1 mRNA suggested that Spec 1 mRNA stability gradually increased throughout development.
    Additional Material: 6 Ill.
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  • 3
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The guinea pig soleus muscle is a convenient model for the study of slow-twitch intermediate (STI) fiber ultrastructure because it is composed entirely of fibers of this class. Such fibers were compared with fast-twitch red (FTR) and fast-twitch white (FTW) fibers from the vastus lateralis muscle.FTW fibers are characterized by small, sparse mitochondria, a narrow Z line and, an extensive sarcoplasmic reticulum arranged primarily in longitudinal profiles at the A band and with numerous expansions at the I band. Abundant mitochondria with a dense matrix and subsarcolemmal and perinuclear aggregations are typical of FTR fibers. These fibers contain a plexus of sarcoplasmic reticulum at the A band and a less extensive network at the I band. The Z lines are wider (890 ± 74 Å) than those of FTW fibers (582 ± 62 Å). STI intermediate fibers are distinguished from other types by wide Z lines (1205 ± 58 Å), a faint M band, and a less extensive sarcoplasmic reticulum. Compared to FTR fibers, STI fiber mitochondria are usually smaller with less notable subsarcolemmal accumulations.FTW fibers have a more limited capillary supply, rarely contain lipid inclusions, and thus may be restricted to phasic activity. Extensive capillarity, mitochondrial and lipid context, and fast contraction times indicate possible phasic and tonic roles for FTR fibers. STI fibers, characterized by numerous lipid inclusions, extensive capillarity, relatively numerous mitochondria, but slow contraction-relaxation cycles, are morphologically suited for tonic muscle activity.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 45 (1991), S. 374-380 
    ISSN: 0730-2312
    Keywords: lipogenesis ; lipolysis ; protein phosphorylation ; second messenger ; glycogen synthesis ; adipocytes ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The pleiotropic nature of insulin action suggests diverse mechanisms of signal transduction for the hormone. The specific protein phosphatase inhibitor, okadaic acid, is utilized to differentiate metabolic pathways that may be regulated by phosphorylation or dephosphorylation of key enzymes. In H-35 hepatoma cells, okadaic acid inhibits insulin-stimulated glycogen synthesis with an IC50 of 400 nM. In contrast, activation of lipogenesis by insulin is inhibited with an IC50 of 50 nM okadaic acid. The toxin also inhibits stimulation of lipogenesis in these cells by the insulin-sensitive inositol glycan enzyme modulator. In isolated rat adipocytes, insulin-stimulated lipogenesis is also inhibited by okadaic acid with an IC50 of approximately 1,700 nM. The antilipolytic effect of insulin in these cells is more sensitive to okadaic acid, exhibiting an IC50 of 150 nM. Maximal activation of lipogenesis by insulin is dramatically reduced by okadaic acid with no effect on the concentration required for half-maximal activation, whereas the sensitivity of insulin-induced antilipolysis is attenuated by okadaic acid, with no apparent reduction in the maximal effect of the hormone. Taken together, these data suggest that specific phosphatases may be differentially involved in some of the metabolic pathways regulated by insulin.
    Additional Material: 6 Ill.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 144 (1990), S. 216-221 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The effects of hyperthermia (41-43°C) on the membrane potential (calculated from the transmembrane distribution of [3H]tetraphenylphosphonium) and Na+ transport of Chinese hamster V79 fibroblasts were studied. At 41°C, hyperthermia induced a membrane hyperpolarization of log phase cells (5 to 26 mV) that was reversible upon returning to 37°C. The hyperpolarization was inhibited 50% by 1 mM ouabain or 0.25 mM amiloride, an inhibitor of Na+: H+ exchange. Shifting temperature to 41°C increased ouabain-sensitive Rb+ uptake indicating activation of the electrogenic Na+ pump. At 43°C for 60 min, the membrane potential of log phase cells depolarized (20-35 mV). Parallel studies demonstrated enhanced Na+ uptake at 41°C only in the presence of ouabain. At 43°C, Na+ uptake was increased relative to controls with or without ouabain present. At both 41 and 43°C, 0.25 mM amiloride inhibited heat-stimulated Na+ uptake. Na+ efflux was enhanced at 41°C in a process inhibited by ouabain. Thus, one consequence of heat treatment at 41°C is activation of Na+:H+ exchange with the resultant increase in cytosolic [Na+] activating the electrogenic Na+ pump. At temperatures ≥43°C, the Na+ pump is inhibited.
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