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  • 1
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Analytical Biochemistry 152 (1986), S. 39-41 
    ISSN: 0003-2697
    Keywords: immobilized fluorescein-labeled gelatin ; proteinase assay
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Experimental Cell Research 215 (1994), S. 40-50 
    ISSN: 0014-4827
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Medicine
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Developmental Biology 126 (1988), S. 203-211 
    ISSN: 0012-1606
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Comparative Biochemistry and Physiology -- Part B: Biochemistry and 99 (1991), S. 709-712 
    ISSN: 0305-0491
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 154 (1978), S. 39-54 
    ISSN: 1432-0568
    Keywords: Implantation ; Trophoblast ; Endometrium ; Zona pellucida ; Proteinases ; Cat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The light microscopical morphology and proteinase activities are studied during the first phases of implantation in the cat, i.e. at 12, 13 and 14 days post coitum (d.p.c.). Timed matings are used to obtain exact data on the time course of implantation events. Size and shape of the blastocysts and the topographical relationships between trophoblast, zona pellucida and endometrium are studied on cryostat sections. These observations indicate that the zona pellucida is being removed at 12 d p.c. by dissolution which starts at the abembryonic pole and lateral of the embryonic disc. Since the zona has, in spite of the considerable expansion of the blastocyst, a thickness of 8–10 μm at this stage, it must have undergone a process of swelling or material must have been added invisibly. Invasion of the trophoblast into the endometrium begins between 13 and 14 d p. c. and is fully under way at 14 d p. c. Widening of the glandular lumina in the neighborhood of the blastocysts at 12–13 d p.c. indicates an early preimplantation interaction between the blastocyst and the endometrium. Amino Acid Arylamidase (Aminopeptidase) activity is found, in histochemical tests, to be high in the trophoblast but low in the endometrium in all three investigated stages. Proteinase Activity is studied with a highly sensitive gelatin substrate film test. Moderate to medium activity is found in the trophoblast at 12–13 d p.c. Very high proteinase activity is present in the invasion zone at 14 d p.c. Experiments with a large number of specific proteinase inhibitors in vitro and preliminary investigation of pH dependence show that it is mainly due to a cathepsin-B-like endopeptidase. This enzyme can be traced to both the trophoblast and the uterine epithelium. The disintegrating zona pellucida shows, at 12 d p.c., only little gelatinolytic proteinase activity. A trypsin-like endopeptidase as described for the rabbit blastocyst could not be identified with certainty in the cat but there is some indication that it might be present at 12 d p.c. Considerable trypsin-like proteinase activity is found in scattered endometrial stroma cells at all stages. The possible physiological role of the described proteinases in implantation is discussed.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 157 (1979), S. 15-34 
    ISSN: 1432-0568
    Keywords: Blastocyst coverings ; Transformation ; Preimplantation development ; Neozona ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The extracellular coverings which surround rabbit blastocysts are far more complex structures than the zona pellucida of other species. Since previously published views of their composition, structure and identification of the various layers are highly controversial, a detailed investigation of the stages between morula and implantation has been undertaken using both electron microscopical and histochemical methods. Rabbit blastocyst coverings undergo considerable structural and chemical transformation from the early until the late blastocyst stages. Morulae are surrounded by zona pellucida and mucoprotein layer (a highly sulfated, sialic acid-free mucosubstance which is derived from the tubal secretion). In the early blastocyst (i.e. from 31/2 d p.c. on), the zona loses its high content of periodate-accessible vicinal hydroxyl groups (PAS reaction) and protein, and there is morphological evidence for an erosion (particularly from the inside), suggesting enzymatic lysis in addition to mechanical stretching due to the expansion of the blastocyst. The zona pellucida disappears completely at 41/2 d p.c. At the same time, deposition of new material begins in its place. This newly formed layer will be called “neozona”. Until implantation, it increases considerably in thickness, finally representing between 2/5 and nearly 1/2 of the total thickness of the blastocyst coverings. Histochemically, the neozona is characterized as a moderately acid mucosubstance, rich in protein and in periodate-accessible vicinal hydroxyl groups, containing sulfate esters as well as sialic acid. Its chemical composition is in many respects comparable to that of the zona pellucida. At least part of the neozona material may be derived from the trophoblast. This new aspect of the physiology of the preimplantation trophoblast, i.e. its secretory activity, is discussed. The possibility that uterine secretion components are also involved in formation of the neozona (as well as in dissolution of the zona pellucida) is envisaged. Observations suggesting a chemical modification of the inner parts of the mucoprotein layer and impregnation of this layer with sialic acid-containing glycoproteins are also discussed. An additional layer, the gloiolemma, which derives from the uterine secretion, is deposited at the outer surface of the mucoprotein layer after 6 d p.c. At the onset of implantation, i.e. around 7 d p.c., rabbit blastocyst coverings are, therefore, composed of three layers of different origin: neozona, mucoprotein layer and gloiolemma.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 179 (1988), S. 129-134 
    ISSN: 1432-0568
    Keywords: Blastocyst coverings ; Neozona ; Zona pellucida ; Trophoblast ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The neozona is the innermost layer of the complex blastocyst coverings of the rabbit and is located between the mucoprotein layer and the trophoblast. As shown previously the neozona begins to replace the zona pellucida from the 4th day post coitum (d p.c.) on (Denker and Gerdes 1979). In the present study, rabbit blastocyst coverings were checked for regional differences in their composition, comparing the embryonic and the abembryonic pole of the blastocyst, at 5 and 6 d p.c. These two stages were chosen because at 5 d p.c. a complete trophoblast layer is still present at both the embryonic disc (Rauber's layer) and the extraembryonic regions (mural and abembryonic pole trophoblast), whereas at 6 d p.c. Rauber's layer has largely degenerated. Correlation of regional differences in blastocyst coverings structure with presence or absence of an intact trophoblast is taken as suggestive evidence for a role of the trophoblast in the formation or the structural modification of blastocyst coverings components. Blastocysts of both stages were fixed in glutaraldehyde with and without ruthenium red and processed for TEM. The neozona was found to be almost equally well developed in all regions at 5 d p.c. On contrast, at 6 d p.c. (Rauber's layer defective) the neozona is consistently found to be much thinner at the embryonic disc than in the extraembryonic regions where the trophoblast is still intact. This is the first report on regional differences of the structural composition of blastocyst coverings within the same blastocyst. It is interpreted as evidence for a physiological role of the trophoblast in the formation of the neozona. The biochemical nature of the putative trophoblastic factors involved is still to be identified.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 183 (1991), S. 17-27 
    ISSN: 1432-0568
    Keywords: Blastocyst coverings ; Zona pellucida ; Neozona ; Preimplantation development ; Synchrony ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Under physiological conditions the zona pellucida disappears in the rabbit between Day 3 and early Day 4 post coitum (p.c.) and is replaced by a new layer, the neozona. The dissolution of the zona pellucida and the formation of the neozona was investigated in three different experimental approaches, all of them characterized by non-physiological developmental conditions for the embryo: Prevention of embryo migration from the oviduct into the uterus by postcoital (48 h p.c.) tubal ligation, in vitro culture, and asynchronous embryo transfer into uteri of recipient rabbits. Embryos of age 21/2, 3, 4 and 41/2 days p.c. were cultured for 12 to 72 h. The media used for in vitro culture were supplemented with BSA, serum or with uterine secretions that were collected either synchronously or asynchronously to the developmental stage of the cultured embryos. Three-day-old embryos were transferred into uteri of pseudopregnant foster rabbits of either synchronous (Day 3) or asynchronous stages (Day 0, 2, 4, 5, 6) and were recovered 24 to 72 h after transfer. The transformation of the coverings was evaluated by light and transmission electron microscopy. The dissolution of the zona pellucida was greatly disturbed in tube-locked embryos, and in cultured embryos if standard protein supplements (BSA or serum) had been used for in vitro culture. In many cases the zona was still completely preserved after 2 or 3 days in culture, at a time when it normally would have already been replaced by the neozona in vivo. The dissolution in vitro, however, progressed incomparably better if the culture medium had been substituted with synchronous or asynchronous uterine secretions. The formation of the neozona could not be verified in cultured blastocysts. After embryo transfer, the dissolution of the zona pellucida was completed in most cases by 2 days after transfer, irrespective of the recipients' progestational stage. Present results indicate that uterine components are essential for the dissolution of the rabbit zona pellucida. These components appear to be present in the uterine cavity constitutively, i.e. independently of the uterine progestational transformation, and need not be in synchrony with the embryo's developmental stage for dissolution of the zona. Normal formation of the neozona does not take place under the non-physiological developmental conditions of in vitro culture.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Histochemistry and cell biology 21 (1970), S. 17-20 
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung Bei der Verwendung von perjodsäureoxydierten Stärkefilmen (Shear und Pearse, 1963) zum Amylasenachweis können Artefakte entstehen durch alkalische, nicht-enzymatische Hydrolyse in Geweben mit hohem pH. Dies wird anhand von Beobachtungen an Kaninchen-Blastozysten erläutert.
    Notes: Summary In substrate film tests for amylase, periodate-oxidized starch films (Shear and Pearse, 1963) may produce artifacts, for they undergo spontaneous hydrolysis at slightly alkaline pH levels. This is concluded from observations made on rabbit blastocysts.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Histochemistry and cell biology 87 (1987), S. 517-529 
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In order to monitor changes in the apical cell membrane of rabbit uterine epithelium which are postulated to be a precondition for trophoblast attachment, the marker enzymes: alkaline phosphatase, aminopeptidase M, γ-glutamyl transferase and dipeptidyl peptidase IV were investigated during the periimplantation phase. Endometrium of early pregnancy (implantation chamber, interblastocyst endometrium; 5–8 days post coitum, d p.c.) was compared with specimens obtained at hCG-induced pseudopregnancy (p. hCG) to distinguish between membrane changes regulated by maternal plasma steroid hormones and such which might be induced locally by blastocyst-derived signals. All enzymes tested showed their main activity at 5 d p.c./p. hCG. The weakest reaction in this series of stages was generally found at 8 d p.c. (interblastocyst segments) or at 8 d p. hCG. In contrast to the rest of the epithelium, the implantation chamber retained high activity of dipeptidyl peptidase IV, and the activity of alkaline phosphatase even raised here again at 7 and 8 d p.c. indicating a direct local influence of the blastocyst on the luminal epithelium. The results suggest that 1) considerable changes occur in the composition of the apical plasma membrane of the uterine epithelium when the endometrium enters the “receptive state”, 2) the overall trend is towards a loss of apical-type characteristics of this membrane domain and 3) the changes are modulated both systemically (by plasma steroid hormone levels) and locally by signals from the implanting blastocyst.
    Type of Medium: Electronic Resource
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