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E Winterhager

Publications and source records attributed to E Winterhager.

68 records · Page 4Linked to original sources

Apical plasma membrane-bound enzymes of rabbit uterine epithelium. Pattern changes during the periimplantation phase.

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, gamma-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.

Animals↗

Regeneration of the uterine epithelium in later stages of pseudopregnancy in the rabbit. An ultrastructural study.

Morphological changes of the uterine epithelium in later stages of pseudopregnancy in the rabbit have been studied using different morphological methods. The highly proliferated mucosa with numerous symplasms of a pseudopregnant animal returns to the morphology of a nonpregnant animal by apoptosis, moderate necrosis and lytic transformation of symplasms back to typical endometrial cells without desquamation of cells. The first signs of lytic transformation are observed on Day 8 of pseudopregnancy. Enhanced regeneration with apoptosis and lysis of the symplasmic nuclei is observed between Day 14 and Day 16. Full restoration of the epithelium with reappearance of ciliated cells, typical columnar and partly mucified epithelial cells is not completed earlier than Day 24 p. hCG. This epithelium, however, differs clearly from the epithelium of a virgin rabbit due to several residues of epithelial transformation. Thus, from a morphological point of view, pseudopregnancy in the rabbit lasts up to or even longer than Day 28 p. hCG with persisting ultrastructural remnants of the preceding cycle.

Animals↗

Transformation of endometrium and fertility in late stages of pseudopregnancy in the rabbit.

Ovulation was induced in rabbits between Days 14 and 18 of pseudopregnancy by an intravenous injection of hCG. Induction of ovulation from Day 16 onwards led to normal progestational endometrial transformation. In rabbits injected on Day 14 or 15, a normal preimplantation endometrial morphology developed, but not earlier than 7 days after hCG (Day 14/15 + 7). Uteroglobin secretion was advanced during the second pseudopregnancy. After mating or artificial insemination, fertility was greatest on Day 18 of pseudopregnancy. Conception failed on Day 14 and embryo transfers were unsuccessful on Day 14 + 1. Transfers performed on Day 14 + 3, however, led to implantation and offspring, even though endometrial morphology did not correspond to the normal Day 3 preimplantational morphology at the time of transfer. We conclude that endometrial transformation typical of normal pseudopregnancy can be induced by ovulation during the regeneration phase of pseudopregnancy from Day 16 onwards; fertilization and implantation can be achieved as early as Day 15 of pseudopregnancy; an oestrous period with high mating activity and fertility occurs about 3 days later; and Day 14 after hCG represents a limited time of functional change from pseudopregnancy to a fertile uterine cycle in the rabbit.

Animals↗

Diffusion barriers in the vaginal epithelium during the estrous cycle in guinea pigs.

In the present study the permeability barriers of the multilayered vaginal epithelium were examined using tracer perfusion techniques, freeze-fracture and thin sectioning. During diestrus and proestrus the upper layers of mucified epithelial cells exhibit tight-junctional belts, which restrict tracer molecules such as lanthanum and horseradish peroxidase. When the highly mucified cells begin to degenerate toward the end of proestrus the underlying epithelium is already keratinized as typical for estrus. The keratinized epithelial cells have a tight-junctional network that joins the basal plasma membranes with the apical membranes of subjacent cells and blocks paracellular diffusion of the tracer molecules. During conversion of the cornified epithelium to a mucified epithelium in metestrus the intercellular space of the epithelium is stained by tracer molecules even though tight-junctional belts can be observed. These results indicate that during cyclic changes of the vaginal epithelium tight junctions can, in general, be considered for the restriction of paracellular diffusion. In metestrus, however, junctions become functionally leaky although they remain morphologically intact. Intercellular lipids, which are normally common in cornified epithelia, are extremely rare and cannot constitute an effective barrier to diffusion in the vagina of the guinea pig. The significance of a strategy that bases the regulation of the permeability on tight junctions rather than on intercellular lipids is discussed.

Animals↗

The fetal vascularisation of term human placental villi. II. Intermediate and terminal villi.

Vessel arrangement and vessel structure of the intermediate and terminal villi of 50 human normal term placentas have been studied by means of semithin histology, three-dimensional reconstruction of serial sections as well as scanning electron microscopy of vessel casts. The reliability of the methods applied has been checked by a morphometrical comparison of the luminal diameters obtained. The mature intermediate villi are characterized by the presence of 1 to 2 terminal arterioles as well as 1 to 2 postcapillary venules, and a few moderately coiled, mostly narrow capillaries, some of which belong to the so-called paravascular network. The remaining capillaries are continuous with the capillary loops of the terminal villi. The fetal vessels of the terminal villi are represented by capillary loops only, parts of which are sinusoidally dilated, reaching diameters up to 50 micrometers. Depending on the method, the mean vessel diameter of the terminal villi is 12.3 (vessel casts) or 14.5 micrometers (semithin sections). The capillaries of the terminal villi are arranged in such a way that 3 to 5 terminal villi are supplied by the same, multiply coiled capillary loop. The average capillary length of the paravascular net is 1,000 to 2,000 micrometers, that of the terminal villus capillary loops 3,000 to 5,000 micrometers. The extent of sinusoidal dilation rises with increasing capillary length, indicating that the main functional importance of the sinusoids is the reduction of blood flow resistance.

Blood Vessels↗

The fetal vascularisation of term human placental villi. I. Peripheral stem villi.

The fetal vascularisation of the most peripheral three generations of stem villi has been studied by means of semithin serial sectioning and scanning electron microscopy of vessel casts in 50 human term placentas. The vessel types have been classified according to the structure of the vessel walls. Stem villi with a diameter of 200-400 microns are characterized by one smaller artery and one small vein, some arterioles and venules and capillaries of the paravascular net. Stem villi of about 150 microns diameter contain arterioles and muscular venules besides the paravascular capillary network. The last generation of stem villi measuring 80 to 100 microns in diameter exhibit a terminal arteriole and a collecting venule as well as up to ten paravascular capillaries. The luminal width of the arterial and venous stem vessels is considerably smaller than described for other vascular beds. This may partly be due to postpartal vascoconstriction. The capillaries of the paravascular net normally to not show sinusoidal dilation. They are arranged as long, hairpin-like, poorly branched loops connecting the arterial and venous stem vessels to each other.

Arteries↗

Membrane events involved in fusion of uterine epithelial cells in pseudopregnant rabbits.

During pseudopregnancy in the rabbit some uterine epithelial cells undergo conversion into symplasmata. This event serves as a model for studies of membrane apposition, fusion and fission of the lateral membranes with the use of different ultrastructural techniques. Apposition of lateral membranes occurs by means of proliferation of the tight-junctional belt and macular tight junctions. Membrane fusion is characterized in freeze-fracture replicas by continuously running fracture planes between neighboring membrane leaflets of epithelial cells, in general without reorganization of the particles. It is suggested that the reorganization of particles as well as the blebs or vesicles of smooth membranes, which are occasionally observed, may be artefacts. Membrane fission occurs simultaneously with fusion resulting in irregularly shaped membrane holes on freeze-fracture replicas. These events are rarely seen in thin sections. Staining with tannic acid reveals that only the layers of the plasma membrane are accessible to this agent. The fusion-fission process starts in the lower region of the lateral membranes, whereas the luminal portion with the broad tight-junctional belt remains intact.

Animals↗

Membrane specializations of the cells of the Harderian gland of the rabbit with particular reference to the mechanism of exocytosis.

Membrane specializations of the cells of the red and white lobe of the Harderian gland, especially membrane differentiations involved in exocytosis, as well as granule interactions were investigated by means of the freeze-fracture and thin-section techniques. Gap junctions are numerous between both types of glandular cells. Tight junctions consist of about five strands, loosely arranged and often fragmented. Large secretion droplets of both cell types containing different kinds of lipids are formed by fusion. The adjacent granule bilayers are devoid of intramembrane particles (IMP) at the fusion site. During exocytosis the large secretion droplets of the red-lobe cells often cause the plasma membrane to bulge out extensively into the lumen. The overlying plasmalemma as well as the granule membrane show IMP-free patches. Secretion granules of the white-lobe cells, smaller in volume, deform the plasma membrane only slightly when lying in close apposition. The distribution of IMP on the plasmalemma of the white-lobe cells remains random at the side of impending fusion. Thin sections sometimes reveal, in this stage of exocytosis, a single unit membrane but never a pentalaminar structure. Both cellular components of the Harderian gland, when pretreated identically, show different membrane reactions with respect to IMP-clearing during exocytosis. This leads to the conclusion that IMP-clearing may not be responsible for membrane fusion but for other complex events in the regulation of membrane-to-membrane interactions.

Animals↗

Alterations in intercellular junctions of the uterine epithelium during the preimplantation phase in the rabbit.

In the rabbit, the pseudopregnant uterus has been used as a model for studying alterations characteristic of the preimplantation phase. Alterations in intercellular junctions of the uterine epithelium were investigated during early pseudopregnancy (day 0 to day 6) by means of the freeze-fracture technique. In the uterine epithelium of oestrous females the zonula occludens belongs to the "tight" type of tight junctions. During pseudopregnancy an impressive proliferation of tight junctional belts can be observed. The basal strands proliferate, forming loops perpendicular to the luminal surface, whereas the more or less parallel arrangement of the luminal strands is maintained. At day 4 of pseudopregnancy macular tight junctions begin to develop on the lower portions of the lateral plasmalemma and are extensive by day 6 post hCG. Small gap junctions are infrequent between cells of the uterine epithelium and show no significant changes during the preimplantation phase. The physiological significance of the present morphological observations is discussed in the light of changes occurring during the preimplantation period.

Animals↗

Effect of hyper-and hypoosmotic solutions on the structure of the Astacus retina. A transmission electron-microscopic study.

Alterations of the retinula cells in the retina of the light-adapted crayfish in response to hyper- and hypoosmotic van Harreveld solutions was examined by transmission electron microscopy. Increased osmolarity of the bathing medium to twice that of the physiological value leads to shrinkage of the retinula-cell somata. Microvilli, on the other hand, do not undergo shrinkage. Some other characteristic irreversible changes do, however, take place, including detachment of microvilli from the soma, showing a two- or three-fold increase in diameter, and a concomitant decrease in number, probably due to fusion of microvilli. Prolonged incubation or higher osmolarities (5 isosmol) cause alteration of the microvilli membranes to whorls. Structural changes are often restricted to microvilli stacks evaginating from certain individual retinular cells. The number of affected stacks increases depending on the duration of incubation or the osmotic pressure. Hypoosmotic solution (0.5 isosmol) also induces an increase of microvillar diameters and a concomitant reduction in number of microvilli per stack. Exposure to a 20% solution of glycerol causes destruction of the rhabdom structure and the formation of whorls from microvillar membranes. The present findings suggest that the structure of the microvilli is stabilized by an axial cytoskeleton.

Animals↗

Cell-cell-communication during placental development and possible implications for trophoblast proliferation and differentiation.

Since direct cell-cell-communication plays a crucial role in the coordination of proliferation and differentiation processes during development we have focused on the expression patterns of gap junctions and their functional implication in the human placenta. The gap junction protein connexin40 (Cx40) is expressed in the proximal extravillous trophoblast of cell islands and columns. In accordance with these observations, isolated trophoblast cells from first and second trimester placentae and choriocarcinoma cells (Jeg-3) reveal Cx40 expression. This channel is not only characteristic of the trophoblast cells along the invasive pathway but also of endothelial cells. To elucidate the functional role of this channel for proliferation and invasion, the non-coupled Jeg-3 cells have been transfected with Cx26, Cx40 and Cx43, respectively. In contrast to Cx40, the Cx26 channel was more potent in reducing proliferation and inducing differentiation indicated by hCG-beta secretion. Using the nude mouse model to study invasion properties of choriocarcinoma cells, we demonstrated that malignant trophoblast cells were able to invade host vessels and to replace endothelial cells. Upregulation of endogeneous connexin genes in tumours grown in nude mice enforces further experimental strategies to investigate the importance of the different channels to fake the cell biological program of endothelial cells.

Animals↗

Expression pattern of integrin adhesion molecules in endometriosis and human endometrium.

Integrins are cell adhesion molecules that undergo cell-specific dynamic changes during the normal menstrual cycle in the human endometrium. Here, using immunohistochemistry, we have investigated the expression pattern of the integrins alphav, alpha2beta1, alpha3beta1, alpha3, alpha6, beta1, beta2 and beta3 in the human ectopic endometrium of 30 patients and in nine cases in the corresponding eutopic endometrium. The biopsies were obtained during the early or late follicular phase (25 cases), during the corpus luteum phase (four cases) and in one case after 6 months' treatment with a gonadotrophin releasing hormone (GnRH) agonist. The integrin expression was independent of the ovarian steroid situation at the time of biopsy. The integrin alpha6 was expressed in all endometriotic and endometrium samples. The integrin alpha3 was absent in all endometrium tissues of patients with endometriosis. However, the corresponding endometriotic lesions re-expressed this adhesion molecule in 15 cases. No change in integrin beta3 expression pattern could be demonstrated in either endometriotic lesions or endometrium samples, regardless of the menstrual cycle phase. A correlation between serum oestradiol and progesterone concentrations and the expression of the investigated integrins was not observed, thus indicating that these two hormones play a minor role in the regulation of the cell adhesion molecules examined. Our investigation suggests that endometriosis is a dedifferentiated disease as it expressed different integrins in comparison with the eutopic endometrium, and independently of the hormonal situation. The ability of endometriotic tissues to express integrins may explain the high recurrence rates in patients with endometriosis, as these samples retain their adhesion potency after retrograde menstruation and are thus able to establish cell-cell and cell-matrix interactions with the surrounding peritoneum.

Biopsy↗