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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 138 (1989), S. 154-164 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Suspensions of freshly isolated rat granulosa cells were used to study endocytosis and processing of radioiodinated ovine follicle-stimulating hormone (l-oFSH) and to analyze the dynamics of its receptor. Ovine FSH was iodinated to a specific activity of 26 μCi/μg as determined by radioreceptor self-displacement assays with maximum specific binding to excess membrane receptors of 46%. Radiolabeled oFSH was judged biologically equivalent to the unlabeled hormone since l-oFSH shows saturation-binding kinetics and stimulates steroidogenesis in a similar dose-related manner to unlabeled oFSH. Experiments designed to study the extent and time course of degradation involved continuous exposure of isolated granulosa cells to l-oFSH. Saturation of membrane receptors was achieved within 1.5 h of incubation, and internalization of FSH occurred in a linear manner for up to 6 h. The rate of internalization was equivalent to 2,780 FSH molecules/cell/h. Degradation of FSH became apparent after 6 h of incubation and increased to 86% of total cellular-associated radioactivity at 22 h. FSH degradation was inhibited by 100 μM chloroquine or 0.45 mM leupeptin. The measurement of cell surface l-oFSH binding in the combined presence of 100 μM chloroquine and 0.5 mM cycloheximide was unchanged for up to 22 h of incubation. This and other receptor binding data suggest that there is no reutilization of FSH receptors. Scatchard analyses of 4°C binding assays on intact cells indicated that a two-site model best fit the data with association constants of K11 = 1.44 (±.42) × 1010 and K12 = 4.35 (±.91) × 108. Receptor binding and activation studies for progesterone production yielded ED50S of 270 pM and 7.7 pM, respectively, and also indicated that 20% receptor occupancy is sufficient to stimulate maximal progesterone production. We conclude that after the initial binding event, FSH is endocytosed very slowly and is subsequently shuttled to the lysosomal compartment for degradation. The retarded rate of endocytosis may relate to novel pathways of hormone processing.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 223 (1989), S. 387-392 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The possibility that ovarian steroids may participate in the inhibition of meiosis has not been rigorously examined. Since progesterone levels are extremely high in follicular fluid prior to ovulation, we tested the possibility that this steroid may be involved in oocyte maturation. To this end, we collected follicular oocytes and cultured them in the presence of dibutyrl cAMP (Bt2), progesterone, and/or the progesterone antagonist RU486 and assessed maturation evidenced by germinal vesicle breakdown (GVBD). Denuded oocytes or cumulus masses collected in the presence of 1 mM Bt2 and subsequently cultured in 25 μM progesterone did not undergo GVBD. However, denuded oocytes and cumulus masses collected in the presence of progesterone and not Bt2 did undergo GVBD (93%). Concentrations of Bt2 (150 μM) that would not inhibit GVBD were inhibitory when used in the presence of progesterone (1-25 μM). Competition experiments using increasing concentrations of the progesterone antagonist RU486 (1-100 μ) did not block the ability of progesterone to enhance the activity of Bt2. We conclude that progesterone alone does not block GVBD; however, in the presence of low concentrations of cAMP it is extremely effective in blocking GVBD. The synergistic activity of progesterone does not appear to be mediated by the progesterone receptor. The data suggest that progesterone and cAMP may operate cooperatively to inhibit meiosis in the ovarian follicle.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 194 (1979), S. 125-141 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The morphology of early postimplantation mouse egg cylinders was studied using light and electron microscopy. Implantation sites at seven, seven and one-half and eight days of gestation were dissected from the myometrium and whole implants, including both decidua and egg cylinders were processed for electron microscopy. Pre-primitive streak egg cylinders were composed of two germ layers, a tall columnar ectoderm and an outer visceral endodermal layer. Ectodermal cells demonstrated large oval nuclei and an organelle sparse cytoplasm except for many free polyribosomes. The visceral endodermal layer was composed of two cell populations. One visceral endodermal cell type observed was tall columnar in shape and appeared absorptive as demonstrated by many microvilli, pinocytotic profiles and lysosomal granules. This population was confined to extraembryonic regions of the egg cylinder. The second visceral endodermal cell type, squamous in shape, evidenced only a few microvilli, pinocytotic profiles and lysosomal granules. This population was confined to the embryonic region of the egg cylinder. Concurrent with the formation of the primitive streak an increased number of cellular junctions and nuclear pores became evident in the ectoderm. Mesodermal cells were large and stellate-shaped exhibiting many filapodia which made contact with adjacent mesodermal elements. Later the cephalic region of the primitive streak proliferated resulting in the migration of wedge-shaped mass of cells, the head process. At the most ventral extremity of the post-primitive streak egg cylinder the cells of the head process became intimately associated with the ectoderm by areas of focal contact and gap junctions.
    Additional Material: 21 Ill.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    American Journal of Anatomy 161 (1981), S. 101-114 
    ISSN: 0002-9106
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: After 3 days in culture, rat ovarian granulosa cells assume a flattened epitheloid organization. Ca+2 and Mg+2 deprivation results in cellular rounding which is reversible and was monitored by phase-contrast time-lapse cinematography. Concomitant with the shape change is a dispersion of the structural proteins actin and α-actinin. The arrays of large actin-containing bundles (stress fibers) are converted to a diffuse network as observed by electron microscopy. Alpha-actinin, which was observed by immunocytochemistry to be in a periodic array along the actin bundles, is disrupted also and redistributed in the periphery of the cell upon rounding. Measurements made of the culture medium during the rounding process indicate that there is a loss of Ca+2 and Mg+2 from the cell interior. These data led us to speculate that Ca+2 and/or Mg+2 are necessary in order to maintain the integrity of stress fibers and/or restrict the movement of α-actinin anchoring sites within the membrane.
    Additional Material: 13 Ill.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    American Journal of Anatomy 167 (1983), S. 395-404 
    ISSN: 0002-9106
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Ovarian granulosa cells grown on glass coverslips were split by a “sandwich” technique. Using this technique we describe a complex filamentous network in the cytoplasm of cultured granulosa cells that was composed of a branching and anastomosing lattice of filaments 20-40 nm in diameter. Since filament identification impossible on the basis of size, split cells were decorated with S-1 fragments of rabbit skeletal muscle myosin. It was readily apparent that the major constituent of the filamentous lattice was actin. Actin was organized in large bundles in which individual filaments were longitudinally aligned. Actin was also observed organized in a loose network throughout the remainder of the cytoplasm. Actin appeared to be intimately associated with organelle and plasma membranes. Coated pits were also a site of actin-filament interaction. Filament polarity was generally away from the membrane with which filaments were associated.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    American Journal of Anatomy 178 (1987), S. 204-213 
    ISSN: 0002-9106
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: We have evaluated the distribution of mitochondria and acidic organelles using, respectively, the specific vital fluorescent dyes rhodamine 123 and acridine orange during preimplantation embryonic development in the mouse. Under conditions used to visualize organelles in living embryos, staining with either dye was found to have no effect on either the rate or extent of in vitro development of five- to eight-cell embryos up to the blastocyst stage. Mitochondria were randomly distributed throughout the cytoplasm and located around nuclei in blastomeres of uncompacted embryos. During compaction, mitochondria initially reorganized to the blastomere cortex; however, these organelles were later confined to the perinuclear region in the trophectoderm (TE) of expanded blastocysts. Acidic organelles were randomly distributed in the cytoplasm of uncompacted embryos, but following compaction, they were concentrated in cortical and perinuclear locations. Moreover, in TE cells of expanded blastocysts, acidic organelles were found exclusively in a tight perinuclear pattern. Microtubules and microfilaments in TE cells were localized in fixed embryos stained with antitubulin antibodies and rhodamine phalloidin, respectively; these structures were found primarily in the cortical cytoplasm at areas of cell-cell contact and secondarily in a perinuclear location. Thus mitochondria and acidic organelles undergo stage-specific redistributions from a diffuse or cortical pattern at the eight-cell stage to a tight perinuclear localization in the TE. We conclude that the polarized distributions of some organelles and cytoskeletal proteins during compaction may not be reliable permanent markers of the mature TE.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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