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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 541 (1988), S. 0 
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Biochemical genetics 9 (1973), S. 247-259 
    ISSN: 1573-4927
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract RNA from 6-day preimplantation rabbit blastocysts hybridizes with 1.8% of the nonrepeated portion of the rabbit genome. This indicates that at least the quantitative equivalent of about 60,000 unique base sequences 1000 nucleotides in length is transcribed. RNA from embryos implanted in the uterus (12-day) hybridizes with 2.5% of the nonrepeated DNA. The extent to which these transcripts in early rabbit embryos are unique to this stage of the mammalian life cycle is not known. About 70% of the newly synthesized heterogeneous RNA (hRNA) from 4- and 6-day blastocysts hybridizes with the slow-renaturing or nonrepeated portion of the genome. Since estimates of messenger RNA (mRNA) content of hRNA (Schultz et al., 1973), based on polyadenylic acid content, suggest that about 20% of the hRNA is mRNA, it appears that the majority of the transcripts from nonrepeated DNA in preimplantation rabbit embryos is not mRNA.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Biochemical genetics 13 (1975), S. 663-671 
    ISSN: 1573-4927
    Keywords: lactate dehydrogenase ; isoenzymes ; differentiation ; blastocyst ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Unfertilized eggs and early embryos up to the 2-day (16-cell) cleavage stage of development in the rabbit contain predominantly the most cathodal lactate dehydrogenase isoenzyme made up of A-type subunits. Following early cleavage there is a progressive increase in total LDH activity in the embryo as development proceeds through 4- and 6-day blastocyst stages. This is accompanied by an increase in the amounts of B-type subunits and a concomitant shift in the lactate dehydrogenase isoenzyme electrophoretic pattern toward the anodal isoenzyme types.
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Biochemical genetics 14 (1976), S. 523-525 
    ISSN: 1573-4927
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 10 (1984), S. 45-56 
    ISSN: 0148-7280
    Keywords: blastocyst ; messenger RNA ; heat shock ; recombinant DNA ; actin ; transcription ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have utilized the rabbit 6-day blastocyst as a model system in which to examine the effect of environmental stress on embryonic gene expression. Elevation of the incubation temperature from 37 to 43 °C, exposure to 50 μM sodium arsenite or mechanical injury (resulting in the structural collapse of the 6-day rabbit blastocyst) was found to depress total protein synthesis as well as enhance the synthesis of a 70,000-dalton stress-induced protein. The molecular mass of this stress protein is similar to a heat shock protein (HSP) found in other eukaryotic systems. A recombinant DNA probe consisting of the 5′ end of a mouse gene for a 70,000-dalton HSP hybridized to RNA isolated from heat shocked, sodium arsenite-treated, and mechanically injured blastocysts but not to RNA isolated from control embryos. These results as well as in vitro translation data suggest that the expression of the 70 K HSP is controlled at the transcriptional level. The levels of actin mRNA, as detected by means of a recombinant DNA probe encoding a Drosophila actin gene, did not undergo a major alteration following these different stresses. The relevance of these observations to embryonic cellular homeostatis is discussed.
    Additional Material: 4 Ill.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 4 (1981), S. 25-34 
    ISSN: 0148-7280
    Keywords: fertilization ; pronuclei ; chromatin decondensation ; protamine ; kinase ; glutathione reductase ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In order to derive information about possible mechanisms by which the sperm head is converted into the male pronucleus during fertilization in the rabbit, unfertilized egg homogenate was assayed for two enzyme activities. Protamine was extracted from rabbit sperm, purified, and labelled with [14C] in an in vitro reaction and used as a probe to assay for a protein kinase which could transfer [32P]PO4 from [γ-32P]ATP onto the substrate. A kinase with a pH optimum of approximately 8.0 to 8.5 is described. Assays for the enzyme glutathione reductase were performed using homogenates from eggs or embryos at three early stages of development. Results suggest that oocytes can oxidize 2.58 × 10-6 μmol NADPH per minute per oocyte, unfertilized eggs 5.16 × 10-7 μmol NADPH per minute per ovum, and 20- to 24-hour postcoitus fertilized eggs 2.30 × 10-6 μmol NADPH per minute per ovum. The relevance of these observations to male pronuclear formation is discussed.
    Additional Material: 2 Ill.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 35 (1993), S. 414-420 
    ISSN: 1040-452X
    Keywords: Embryonic development ; Fetal development ; IGFBPs ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The temporal patterns of expression of genes encoding insulin-like growth factor (IGF) ligands and receptors during very early development have been investigated in several laboratories in several different mammalian species. Both reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemical techniques have been used to identify the time of appearance of gene transcripts or end-products. In preimplantation mouse embryos, IGF-II ligand and receptor gene activity is detectable as early as at the two-cell stage, the time when transcription from the embryonic genome is activated, but receptors for insulin and IGF-I are not detectable until the compacted eight-cell stage. Transcripts for insulin or IGF-I are not detectable in preimplantation mouse embryos, although the ligands are present in the reproductive tract. The pattern of IGF gene expression is not, however, identical in all mammalian species. In cow embryos, for example, transcripts for IGF-I and IGF-II ligands and receptors and insulin receptors have been detected at all stages of preimplantation development from mature oocyte to blastocyst (Watson et al., 1992). Attempts to quantitate transcript abundance in these early embryos are in progress in our laboratory. In the preimplantation mouse embryo, transcripts for several different IGF-binding proteins (IGFBP-2, -3, -4, and -6) have been detected by RTPCR procedures. In addition, transcripts for IGFBPs have been identified in RNA derived from cumulus cells, the ovary, the oviduct, the uterus, and the decidua. These findings suggest that the interactions of IGF ligands and receptors in preimplantation development might, indeed, be modulated by IGFBPs. Approaches to function of IGFs in preimplantation embryos have involved analysis of the stimulatory effects on metabolism and cell division when IGFs are added exogenously to embryos in culture in simple defined medium (for example, see Harvey and Kaye, 1991) or observations in the reduction in rate of development following interference with IGF expression (see Rappolee et al., 1992). In general, members of the insulin and IGF gene family of polypeptides have been shown to lead to an enhahancement of development in vitro in both laboratory and domestic mammalian species. © 1993 Wiley-Liss, Inc.
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 31 (1992), S. 87-95 
    ISSN: 1040-452X
    Keywords: Mammalian development ; mRNA phenotyping ; RT-PCR ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The sensitive technique of mRNA phenotyping with the reverse transcription-polymerase chain reaction was employed to determine the patterns of gene expression for several growth factor ligand and receptor genes during bovine preimplantation development. Several thousand bovine embryos encompassing a developmental series from one-cell zygotes to hatched blastocysts were produced by the application of in vitro maturation, fertilization, and oviductal epithelial cell embryo coculture methods. Transcripts for transforming growth factor (TGF-α) and platelet-derived growth factor (PDGF-A) are detectable in all preimplantation bovine stages as observed in the mouse. Transcripts for TGF-β2 and insulin-like growth factor (IGF-II) and the receptors for PDGF-α, insulin, IGF-I, and IGF-II are also detectable throughout bovine preimplantation development, suggesting that these mRNAs are products of both the maternal and the embryonic genomes in the cow, whereas in the mouse they are present only following the activation of the embryonic genome at the two-cell stage. In contrast to the mouse embryo, IGF-I mRNA was detected within preimplantation bovine embryos. Basic fibroblast growth factor (bFGF) is a maternal message in the bovine embryo, since it is only detectable up until the eight-cell embryo stage. Bovine trophoblast protein (bTP) mRNA was detectable within day 8 bovine blastocysts. As was observed in the mouse, the transcripts for insulin, epidermal growth factor (EGF), or nerve growth factor (NGF) were not detectable in any bovine embryo stage. Analyses of this type should aid the development of a completely defined culture medium for the more efficient production of preimplantation bovine embryos.
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  • 9
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 31 (1992), S. 231-240 
    ISSN: 1040-452X
    Keywords: Maternal mRNA ; mRNA processing ; Mammalian development ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: This study describes the localization of the U2 small nuclear RNA (snRNA) and the major U snRNA group ribonucleoproteins (snRNPs) during bovine preimplantation development. In vitro maturation, fertilization, and oviductal epithelial cell coculture methods were employed to produce several developmental series totalling over 2,000 preimplantation-stage bovine oocytes and embryos. These oocytes and preimplantation embryos were processed for in situ hybridization, immunofluorescence and Northern blotting methods. The U2 snRNA and the major U group snRNPS were localized initially over the germinal vesicle (GV) of preovulatory oocytes but following GV breakdown were released throughout the ooplasm. They subsequently reassociated with both pronuclei during fertilization. From the two-cell to the blastocyst stages, the U2 snRNA and U snRNPs were localized to the interphase nucleus of each blastomere. The levels of U2 snRNA throughout bovine preimplantation development were determined by probing a Northern blot containing total RNA isolated from the following preimplantation bovine embryo stages: one to two cell, eight to 16 cell, early morula (〉32 cell), and late morula/early blastocysts. The levels of U2 snRNA remained constant between the one-cell and eight-to 16-cell bovine embryo stages but increased 4.4-fold between the eight- to 16-cell stage and the late morula/early blastocyst stages. The results suggest that a maternal pool of snRNAs is maintained in mammalian preimplantation embryos regardless of the duration of maternal control of development.
    Additional Material: 6 Ill.
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  • 10
    ISSN: 1040-452X
    Keywords: Reverse transcription ; Polymerase chain reaction ; Embryogenesis (mouse) ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The expression of genes for insulin and insulin-like growth factors (IGFs) and their receptors was examined in early postimplantation mouse embryos and differentiating F9 embryonal carcinoma cells using mRNA phenotyping. Messenger RNA phenotyping involves the reverse transcription of RNA followed by amplification of specific target cDNA sequences using the polymerase chain reaction (PCR). The identities of the resulting PCR fragments were confirmed using at least two of the following methods: (1) size determination by agarose gel electrophoresis, (2) the presence of diagnostic restriction sites, (3) hybridization with radio-labeled cDNA probes, (4) sequencing of the PCR fragment. Transcripts for insulin receptors, IGF-I receptors, and IGF-II receptors were detected in RNA samples from day 7.5 to day 9.5 mouse embryos and in F9 cells, although the level of insulin receptor mRNA in F9 cells was very low. Transcripts for both IGF-I and IGF-II ligands were also detectable in the embryo and F9 RNA samples, but transcripts for insulin ligand were undetectable in either set of material. The results suggest that insulin does not act as a paracrine or autocrine growth factor in early postimplantation embryos or F9 cells but that both embryos and F9 cells have the potential to respond to exogenous (e.g., maternal) sources of insulin. Both IGF-I and IGF-II could act as paracrine or autocrine growth factors, and IGF-II is the more abundant growth factor in differentiating F9 cells.
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