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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
    Medical electron microscopy 32 (1999), S. 67-78 
    ISSN: 1437-773X
    Keywords: Key words Electron microscopy ; Human yolk sac ; Yolk stalk ; Vitelline duct ; Human embryo ; Human development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Organogenesis occurs during the first 8 weeks of human embryonic development; in consequence, early human growth and development take place before and in the absence of fully developed internal organs. During this period, normal development depends on several factors, but two are imperative: nutrition and a functional transport system for the distribution of nutrients and for waste disposal. The yolk sac (YS), a highly differentiated adnexal organ, is known to accomplish this fundamental task during early pregnancy. In this review, we summarize our contribution to the understanding of early human embryology, focusing interest on analysis of the morphofunctional link that is established between the human embryo and the YS during the embryonic period. Embryos were collected from the gestational sac after salpingectomies performed on patients with singleton pregnancies occurring in the fallopian tube. Samples of YS were taken from 20 human embryos at Carnegie stages ranging from 12 to 20. The age of the embryo was estimated from data of the patient's last menstrual history and confirmed from crown-rump length measurements and morphological characteristics of the specimen. The samples were fixed in 3% glutaraldehyde and then postfixed in 2% osmium tetroxide and prepared for light, transmission, and scanning electron microscopy according to conventional techniques. The samples were examined with a Philips 301 EM and an S-4000 Hitachi field emission SEM. The yolk stalk, and the YS wall with its corresponding endodermal, mesenchymal, and mesothelial layers, were analyzed. In accordance with their morphological features, the endodermal cells are equipped with organelles to fulfill several functions that are expressed in absorption from the vitelline cavity via microvilli present into the outer cell surface, in secretion to the extracellular space, and in the synthesis of numerous proteins which are transported by the bloodstream to the embryo. The mesothelial surface is provided with cell-surface differentiation that promotes a protective coat to prevent damage from compression or friction of the YS wall against the amnios, umbilical cord, and chorionic cavity wall during growth. The mesenchyme is the main site for blood vessel formation and gives rise to a network that provides the embryo with nutrients and a means of waste disposal. A critical analysis of the role of the endodermal vesicle in the production of fluid that is accumulated in the YS, and of the role that the vitelline duct play in the exchange function between the YS and intestinal tract, is presented. We have demonstrated that the vitelline duct is not functional after week 5 because of the closure of its lumen. This finding is discussed with reference to the biological meaning of the vitelline duct and its functional period of activity, and its possible role in the physiology of exchange during the embryonic period is assessed.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-7330
    Keywords: blastocyst ; coculture ; human granulosa cells ; human embryos ; IVF implantation failures
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Purpose: Our purpose was to determine the effects of thecoculture of embryos on human granulosa cells (GCs) inpatients in the first cycle of IVF-ET treatment and in patientswith repeated implantation failures and to investigate thepresence of specific proteins in a 48-hr GC conditionedmedium and the GC ultrastructural characteristics. Methods: Eighteen patients with tubal or idiopathicinfertility were enrolled in this study: 7 patients (Trial 1) were inthe first cycle of IVF-ET treatment and 11 patients (Trial 2)had repeated implantation failures (one to five). Embryosfrom each patient were cocultured randomly either onhomologous granulosa cells or on a conventional culturemedium. Results: At the end of the coculture period (day 5 or 6),50% of the embryos (Trial 1) reached the blastocyst stage,with respect to 35% in Trial 2. The pregnancy rate perretrieval was 14.2 and 9%, respectively, in Trial 1 and inTrial 2. Many conditioned media showed proteins of 24–29kDa. and some of them showed additional proteins of 90kDa. The ultrastructural analysis of GCs showed healthy,metabolically active, protein-synthesizing, and mostlysteroidogenic cells. Conclusions: GC cultures improve embryo development butnot pregnancy rates both in Trial 1 and in Trial 2.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1573-7330
    Keywords: oocyte ; zona pellucida ; in vitro fertilization ; scanning electron microscopy ; gamete interaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The structure of the zona pellucida and the early interactions between human oocytes and spermatozoa were investigated in an in vitro fertilization program. Thirty-five mature (preovulatory) oocytes, 10 immature oocytes lacking a germinal vesicle, and 11 atretic oocytes which had not undergone fertilization at 10–20 hr after insemination were studied by light and scanning electron microscopy. Observed through employment of these techniques, the zona pellucida showed two basically different patterns: a mesh-like, spongy structure having wide and/or close meshes; and a compact, smooth surface. The smooth-surfaced zona was most commonly seen in the cultured oocytes belonging to the immature and atretic groups. These observations seem to show that the spongy appearance of the zona pellucida is related mainly to oocyte development and maturity. In this study, greater numbers of penetrating spermatozoa were noted on oocytes showing the mesh-like zona, in contrast to the presence of a few sperm flattened against its surface or the frank absence of sperm associated with oocytes having the more compact, smooth zona. It is likely that the condensation of the outer aspect of the zona pellucida causes a disorientation of sperm-binding sites, which would probably result in markedly reduced binding and penetration capacity with spermatozoa. These changes might ultimately lead to impairment of in vitro oocyte fertilizability.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-0878
    Keywords: Key words: Ovary ; Vascularization ; Vascular corrosion casts ; Scanning electron microscopy ; Countercurrent mechanism ; Rabbit (New Zealand white)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Ovarian angioarchitecture was studied by scanning electron microscopy of vascular corrosion casts in estrous, pseudopregnant (stimulated with human chorionic gonadotropin) and pregnant rabbits. In all samples, the proper ovarian branch of the ovarian artery (ramus ovaricus) entered the ovarian hilus near the caudal pole of the organ and ran parallel to the major axis of the hilus. The extraovarian venous drainage was formed by several vessels emptying into a distal large vein. The ramus ovaricus exhibited various degrees of coiling and branched in the medulla. The coiling of the ramus ovaricus and its ramifications were maintained in all samples. A venous meshwork and/or flat vein branches closely enveloped the arterial coils found in the hilus and outer medulla. At this level numerous arteriovenous contacts were demonstrated in all samples. The coiled arteries, prior to entering the ovarian cortex, supplied several small peripheral follicles which were drained by the hilar veins. In the cortex the coiled arteries branched in numerous thin, straight or slightly undulated arterioles which supplied developing estrous follicles and pseudopregnant corpora lutea. The arterioles supplying the pregnant corpora lutea were long, large and tightly spiraled. The venous drainage followed the modifications of the arterial supply. These data demonstrate that ovarian cycle and pregnancy induced significant changes in the cortical vessels, which adapted their structure to the temporary functional needs of the recruited follicles or corpora lutea. Hilar and medullary vessels have permanent structures that may represent morphological devices for (a) a continuous control of the blood flow (spiral arteries) and (b) a local recirculation of endocrine products (arteriovenous contacts) comparable to the ”countercurrent mechanism” previously shown to operate in ovaries of other species, but not yet found in rabbits.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 226 (1982), S. 493-510 
    ISSN: 1432-0878
    Keywords: Ovarian cords ; Surface epithelium ; Fetal germ cells ; Mesonephros ; Ultrastructure ; SEM
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ovarian surface and associated germ cells have been studied in human fetuses from 12 weeks of age until near term, using light, TEM and SEM techniques. The surface epithelium and related cords proliferate extensively, especially at midterm. The cords in the ovarian cortex appear to be linked with ingrowths from the surface epithelium, and both structures have a common basal lamina. Germ cells are always interspersed among the somatic cells of the surface epithelium and associated cords. These results indicate that both the proliferating cords and surface epithelium may contribute to the formation of early follicles. Furthermore, the occurrence, of elements having some of the features of primitive steroidogenic cells in the regions of cordsurface epithelium continuity, suggests that both structures (surface epithelium and cords) contribute somatic cells, which in addition to becoming granulosa cells, might also contribute to the provision of primitive interstitial cells. Gonocytes tend to migrate through the developing ovarian tissue towards the surface where they become extruded into the peritoneal cavity. This phenomenon might contribute to the reduction in the number of germ cells at birth and parallels the atretic processes within the ovary.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-0878
    Keywords: Zona pellucida ; Ovarian follicles ; Atresia ; Ultrastructure ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The present study provides further details on the fine-structural three-dimensional architecture of the zona pellucida (ZP) in growing and atretic follicles of mice by use of ruthenium red in combination with the detergents Triton X100 and saponin. These detergents were used for extraction of the “soluble” fraction of the zonal proteins in an attempt to expose the “structural” zonal glycoproteins, which in turn can be viewed as minute three-dimensional networks upon transmission- and scanning electron-microscopic examination. By use of these methods, the ZP of growing follicles appeared to be formed by interconnected filaments which also bind to globular structures building up a three-dimensional lattice. In contrast, the ZP of stage I as well as other (II and III) stages of atretic follicles showed a structure characterized by the presence of closely packed granules connected with short filaments to form a close-mesh reticulum. This structural change of the ZP, which in the present study is also associated with the disappearance of “gap junctions” within the granulosa and cumulus cell population, might represent one of the early events involved in the onset of atresia. These changes, most probably depending on an altered secretory activity of both oocytes and follicle cells, might lead to a degradation of the ZP network structure and to its subsequent increased density (condensation). All these morphodynamic events eventually contribute to a sequestration of the oocyte in the early stage of atresia.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 240 (1985), S. 519-528 
    ISSN: 1432-0878
    Keywords: Polycystic mouse ovaries ; Hormone treatment ; Morphometry ; Gap junctions ; Ovary
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Morphological alterations induced by dehydroepiandrosterone (DHA) were studied in polycystic mouse ovaries (PCO). Treated mice showed ovulatory failure and cystic changes; cysts and follicles in various stages of growth and atresia were present although corpora lutea were absent. The levels of testosterone, dihydrotestosterone, 3α- and 3β-androstanediol, estrone and androstenedione increased, whereas estradiol was not detectable. The ultrastructure of granulosa cells in healthy and atretic follicles was similar to that of control animals, although the membrana granulosa in cysts was reduced to a monolayer of flattened cells. The theca interna of healthy and atretic follicles and ovarian cysts showed ultrastructural signs of abnormal steroidogenic stimulation. No significant differences (0.7〈P〈0.8) were found between the extensive surface area of gap junctions of healthy follicles of control and DHA-treated animals. On the P-face of granulosa cells of large healthy follicles, meandering strands of tight junctional particles were observed; their average length was significantly longer than those in healthy follicles of control animals (P〈0.001). This increase was probably related to the large amounts of androgens present in the treated animals. Theca interna cells possessed small gap junctions; no significant differences (P〉0.9) in gap-junction surface area were observed between DHA-treated and control animals. These results suggest that the size of gap junctions is probably unrelated to the steroidogenic activities of theca cells.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 241 (1985), S. 275-281 
    ISSN: 1432-0878
    Keywords: Pituitary cleft ; Supramarginal cells ; Ventricular system ; Hypophysis ; Supraependymal cells ; Hypothalamus ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Unusual cells lying above the marginal cellular layer of the rat pituitary cleft were studied by SEM and TEM. These cells — from their location termed supramarginal cells — have a characteristically irregular cell body from which arise a number of long and thin branched processes ending among the microvilli and cilia of the marginal cells delimiting the anterior and posterior walls of the cleft. Some supramarginal cells are star-shaped elements with thin extensions, others have a triangular or spindle-shaped body from which emerge long ameboid processes with fibril-like projections. Miniblebs, miniruffles, occasional veils and short microvilli extend over the surface of these elements. Supramarginal cells are very similar to the “Kolmer epiplexus cells” originally found on the choroid plexus or other areas of the third ventricular wall where they are known as “supraependymal cells”. Present evidence suggests that supramarginal cells of pituitary cleft might have phagocytic properties and an hematogenous origin as monocytes and, as such, closely resemble Kolmer epiplexus cells of brain ventricles. Others might arise from “folliculo-stellate cells” or closely related marginal cells once they become free into the pituitary cleft. Supramarginal cells are recognized as motile phagocytes acting as scavengers and possibly regulating the extracellular environment of the cleft and associated adenohypophy-sial tissues. In considering the area where they are located supramarginal cells might assume a specific role in relation to the integrated neuroendocrine function of the hypothalamus-ventricle-hypophysial complex. To this complex the pituitary cleft is closely associated embryologically and anatomically.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1040-452X
    Keywords: Oocyte ; Polypronuclear ovum ; Fertilization ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The human zona pellucida (ZP) and its changes during in vitro fertilization in oocytes at different maturational stages and polypronuclear ova at one- to four-cells stages were studied by transmission electron microscopy (TEM) and correlative scanning electron microscopy (SEM). To define the microstructure of the ZP, its amorphous masking material was removed using a detergent (saponin), and its structural glycoproteins were stabilized with a cationic dye, ruthenium red, followed by osmium-thiocarbohydrazide treatment. These methods allowed in all samples the clear visualization of variously arranged networks of filaments composing the outer and inner surfaces of the ZP. These filaments were straight or curved, 0.1-0.4 μ in length and 10-14 nm thick as seen via TEM or 22-28 nm thick as seen via SEM (the difference in thickness was due to the presence of the metal coating for SEM). The filament arrangement was remarkably different between the inner and outer surfaces of the ZP and among the various stages studied. The filaments of the outer surface of the ZP were basically arranged in “large” and “tight” meshed networks. Mature oocytes and fertilized (polypronuclear) ova had a regular alternating pattern of wide and tight meshed networks of filaments. On the other hand, immature and atretic oocytes displayed almost exclusively a tight meshed network of filaments. The inner surface filaments of the ZP of unfertilized oocytes at any stage were arranged in repetitive structures characterized by numerous short and straight filaments anastomosing with each other and sometimes forming at the intersections small, rounded structures. After fertilization, the inner surface of the ZP displayed numerous areas where filaments fused together. Collectively, these data clearly reveal that oocyte maturation and fertilization in humans are accompanied by changes of ZP filaments arrangement, which may be relevant in the processes of binding, penetration, and selection of spermatozoa. © 1992 Wiley-Liss, Inc.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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