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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 13 (1986), S. 103-114 
    ISSN: 0148-7280
    Keywords: demecolcine ; colcemid ; meiosis ; oocyte ; nuclear migration ; nuclear dissolution ; nuclear centrifugation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Germinal vesicle migration (GVM) and dissolution (GVD) were studied in goldfish oocytes treated with 17-α,20-β-dihydroxyprogesterone (DHP) and/or demecolcine (DE; a colchicine derivative also known as colcemid) in vitro. DE (100 μg/ml) in the presence of DHP, enhanced steroid-induced GVM, after both 24 and 48 hr of incubation and significantly reduced the DHP ED50 value for GVM. Similarly, administration of DE alone elicited a significant, dose-related increase in GVM after 24 or 48 hr of incubation. The presence of DE, either alone or in combination with DHP, was without effect on GVD. The effect of DE was also tested on ooplasmic viscoelasticity in goldfish follicles subjected to a centrifugal force (160g for 1 min). Preincubation (24 hr) of goldfish follicles in DE significantly influenced the direction and the extent of the centrifugally induced GV movement along the axis of centrifugal force in a dose-related fashion. The present results provide support for the hypothesis that cytoskeletal components, such as microtubules that are sensitive to DE, are involved in the mechanism of GVM in goldfish oocytes.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 13 (1986), S. 241-251 
    ISSN: 0148-7280
    Keywords: oocyte ; fertilization ; cortical granule ; pig ; calcium ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The structure of pig oocytes after in vivo and in vitro fertilization and following treatment with the ionophore A23187 with differing levels of calcium are described, with particular reference to the cortical granules.Fertilization in vivo and in vitro resulted in cortical granule exocytosis. Sperm penetration in vivo was more rapid than in vitro and resulted in the dispersal of the cortical granules' contents in the perivitelline space following exocytosis. The contents of the granules remained adjacent to the plasmalemma after in vitro fertilization and suboolemmar vesicles were less numerous than after in vivo fertilization. High calcium levels were necessary to induce the dispersal of the cortical granule contents following treatment with ionophore. The observations are discussed regarding their relevance to the blockage to polyspermy.
    Additional Material: 15 Ill.
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  • 3
    ISSN: 0148-7280
    Keywords: oocyte ; demecolcine ; Colcemid ; meiosis ; germinal vesicle migration ; microelectrode intracellular recording ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Demecolcine (Colcemid; DE), a colchicine derivative, augmented meiosis reinitiation by progesterone in the follicle-enclosed oocyte of the frog, Rana pipiens. Whereas DE treatment alone had a minor stimulatory effect on germinal vesicle dissolution (GVD), this treatment elicited significant germinal vesicle movement (GVM) as evidenced by translocation of the GV to the oocyte surface. The effects of DE on GVM and progesterone-induced GVD were also elicited in oocytes lacking follicle cells or other follicle wall components (type IV follicles), indicating that DE has a direct action on the oocyte itself. DE alone did not alter oocyte membrane voltage (Vm), resistance (Rm), or current (Im) and did not interfere with the changes in these parameters usually elicited by progesterone. After 5 hr incubation of follicle-enclosed oocytes with either DE or progesterone, or combinations of both, the GV could be moved to the animal pole surface with less centrifugal force compared to control follicles. This result suggests that a decrease in ooplasmic viscoelasticity is induced by progesterone, which is mimicked by DE before GVM or GVD normally begins. The results presented here support the idea that DE-sensitive oocyte components such as microtubules are involved in the process of steroid-induced meiosis. These findings provide a physiological basis for future studies of cytoskeletal involvement in the events of meiosis.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 14 (1986), S. 65-73 
    ISSN: 0148-7280
    Keywords: nucleolar ; ultrastructure ; bovine ; oocyte ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Nucleolar ultrastructural changes occurring in vivo in bovine oocytes during follicular growth were analyzed by electron microscopy. The rates of in vitro incorporation of 3H-uridine by oocytes of the same size class were evaluated by autoradiography.One to two large fibrillogranular, vacuolated nucleoli were present in oocytes from small to medium antral follicles 0.5-3 mm in diameter. These oocytes showed intense hnRNA and rRNA synthesis. The homogeneous, agranular nucleoli in oocytes from follicles 3-4 mm in diameter were composed of a compact fibrillar material. This morphological change was accompanied by an impairment of nucleolar transcriptional activity as well as by a decrease in hnRNA synthesis.
    Additional Material: 8 Ill.
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  • 5
    ISSN: 0148-7280
    Keywords: oocyte ; freeze-fracture ; lipid probes ; filipin ; polymyxin B ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Membrane and cytoplasmic changes were observed after in vitro fertilization of hamster oocytes by examining freeze-fracture replicas. The density of intramembranous particles on areas of membrane between microvilli increased following fertilization. Although the intramembranous particle density of microvilli is higher than that on the intermicrovillar membrane of unfertilized eggs, it did not change significantly after fertilization. Cytoplasmic changes in the Golgi complex and mitochondria upon fertilization indicate a change in cellular activity. Lipid binding probes were applied to the oocyte membranes in order to study the distribution of specific lipids before and after fertilization. Probes included the B-hydroxy-steroid complexing molecules, filipin and tomatin, and an anionic lipid binding antibiotic, polymyxin B. Both tomatin and filipin complex with steroids in the P and E faces of the plasma membrane (including the polar bodies), cortical granules and vesicles deeper in the cytoplasm, and the Golgi complex, leaving mitochondria, pronuclei, endoplasmic reticulum, and the majority of vesicles unlabeled. Polymyxin B binding is dependent on its application before or after fixation or in association with EGTA. With its application we detected both minor membrane perturbations of wrinkles and particle redistributions and major perturbations of vesicle fusions, the formation of blebs, and the loss of membrane morphology. Neither the distribution nor apparent quantity of these probes changed overall following fertilization, but this impression does not include specific sites of sperm-egg fusion.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 13 (1986), S. 173-184 
    ISSN: 0148-7280
    Keywords: oocyte ; androgen ; in vitro maturation ; pig ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Recent work has shown that 19-norandrostenedione is a major steroidal component of porcine follicular fluid; however, little is known about its role(s) in the regulation of follicular function. This study was designed to examine the effect of 19-norandrostenedione on porcine oocyte maturation in vitro. Oocyte-cumulus complexes were isolated from medium (3-6-mm diameter)-sized prepubertal pig follicles and incubated for 12 h in medium with or without dibutyryl cyclic AMP ((Bu)2cAMP, 1 mM) with or without testosterone (5 x 10-7 M) or 19-norandrostenedione (5 x 10-7 M). In medium alone, 70.8% of oocytes spontaneously resumed meiosis as evidenced by the occurrence of germinal vesicle breakdown. Oocyte maturation was inhibited by (Bu)2cAMP (44.6% of oocytes matured). Although neither steroid alone affected maturation, both testosterone and 19-norandrostenedione enhanced the effect of (Bu)2cAMP (22.5 and 19.6%, respectively, resumed meiosis).The effects of testosterone and 19-norandrostenedione on (Bu)2cAMP-inhibited oocyte maturation were dose dependent and there was no significant difference between the actions of the steroids. The effect of 19-norandrostenedione was reversible and dependent on the presence of an intact cumulus. Hydroxyflutamide (SCH-16423), a nonsteroidal compound known to block androgen receptors, abolished the effects of both testosterone and 19-norandrostenedione on germinal vesicle breakdown, indicating that the actions of these steroids are truly androgenic.The results of this study suggest that 19-norandrostenedione may be of physiological importance in the regulation of porcine oocyte maturation.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 13 (1986), S. 39-46 
    ISSN: 0148-7280
    Keywords: oocyte ; oocyte maturation ; meiotic competence ; follicular development ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Hypophysectomy of 15-day-old rats (hypox) markedly reduced the normal development of meiotic competence and abolished the development of antral follicles between days 21 and 31 postpartum (pp). Here the correlation among age of the rats, stage of follicular development, and meiotic competence was examined. Administration of pregnant mare's serum gonadotropin (PMSG-3IU) or insertion of an estradiol-17β (E2) capsule to hypox rats induced the development of meiotic competence provided the treatment started after day 20 pp. Hormonal treatments at an earlier age were not effective in inducing meiotic competence in hypox rats. The induction of meiotic competence by PMSG or E2 was associated with an increase in the number of granulosa cells and formation of follicular antrum. The finding that PMSG and E2 failed to induce meiotic competence when administered prior to day 21 pp suggests that the development of meiotic competence is an age-dependent process. When the hormonal treatments commenced after day 21, both follicular development and meiotic competence were induced.
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  • 8
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 7 (1986), S. 21-34 
    ISSN: 0192-253X
    Keywords: Drosophila ; pattern formation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The distribution of the soluble form of NADP+ -dependent isocitrate dehydrogenase (ICDH) was examined in Drosophila melanogaster imaginal discs. Development of the enzyme patterns and the specific transformations of the patterns by homoeotic mutants were studied. ICDH pattern formation was followed in eye-antennal discs and wing discs from the late 2nd instar stage through 3rd instar and 8 hours into prepupal development. The patterns formed gradually in both disc types. The most interesting pattern developed in the eye portion of the eye-antennal disc complex. ICDH distribution as well as staining intensity correlated well with differentiation of the ommatidia. The spatial distribution of ICDH within the discs was under genetic control. The patterns reflected the state of determination of the disc. When the presumptive tissue type was transformed via mutant homoeotic genes to different determinative states, the ICDH pattern likewise transformed to the pattern characteristic of the newly acquired structure.
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  • 9
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 3 (1986), S. 143-155 
    ISSN: 0739-4462
    Keywords: Drosophila ; P elements ; regulatory sequences ; polytene chromosomes ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A new picture of gene and chromosome structure is emerging in Drosophila that differs considerably from that largely derived from polytene chromosome banding patterns and saturation mutagenesis. In particular, gene transfer has enabled us to more clearly limit the functional unit of a number of genes. Gene regulation may be studied at the molecular level with such techniques. The possible complexity of regulatory elements that may pose problems in their analysis is discussed.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 3 (1986), S. 35-46 
    ISSN: 0739-4462
    Keywords: locust ; vitellogenin ; juvenile hormone ; P element ; Drosophila ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In Locusta migratoria, vitellogenin (Vg) is normally produced only in adult female fat body under stimulation by juvenile hormone (JH). This permits study of (a) programming of genes for expression and (b) modulation of expression by JH. From L. migratoria genomic libraries, two Vg genes (A and B) have been cloned. Coding regions encompass 10-12 kbases of DNA, contain introns, and hybridize with 6,300 nucleotide mRNAs. Although the 5'-exons, coding for signal peptides, are similar in sequence, the remaining coding sequences have distinct restriction maps and show no crosshybridization. Upstream DNA from the two genes has some sequence similarity, corresponding to potential regulatory regions. Dot hybridization assays of mRNAs A and B in locust fat body after induction by methoprene show coordinate expression of the two Vg genes and also a third, unidentified JH-regulated gene. In the hope of providing a system for identification of control sequences, P element-mediated transformation has been used to transfer locust DNA into Drosophila. From Vg gene B, a central block was deleted, to give a mini-Vg gene comprising 5' and 3' terminal coding sequences plus upstream flanking DNA. This was incorporated into the P element vector Carnegie 20 and injected into Drosophila embryos. Three transformed fly lines were obtained in which the locust mini-Vg gene was integrated at different chromosomal sites. On Northern blots of fly RNA, however, no expression of the locust sequences could be detected, even though the accompanying rosy gene was expressed. This suggests that the locust regulatory sequences were not recognized in Drosophila, and current effort is directed toward developing a homologous locust transformation system for expression assay of cloned JH-regulated genes.
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