Migration of intravascular trophoblast cells in uterine arteries of the golden hamster: a scanning electron microscopic study.

Intravascular trophoblast (IVT) cells, derived from the trophoblast of the developing hamster embryo, are known to migrate in retrograde fashion into the uterine arteries. There they migrate to a certain point, destroy and replace the endothelial lining, and modify the smooth muscle of the arteries. The dilated vessels that result presumably enhance the flow of blood to the placental exchange area. The morphology of IVT cells in the hamster placenta was investigated by scanning and transmission electron microscopy. Although occasional single migrating cells were observed, the IVT generally appear as sheets of large, contiguous, sometimes overlapping cells that spread over the endothelial surfaces of the uterine central terminal arteries and vascular knot arteries. This process seems to be aided by the appearance of filopodia, which make contact either with other intravascular trophoblast cells or the endothelium. After consolidation, the IVT cells act as a functional part of the vessel lining and are readily distinguished from the surrounding endothelium by their numerous microvilli. The final distribution of the IVT cells is patchy rather than uniform.

Oxytocin-induced prostaglandin release from perifused bovine caruncular and intercaruncular endometrial tissue on days 20, 30 and at first estrus postpartum.

Twenty-two multiparous Brahman x Hereford F1 cows were utilized to determine the effect of oxytocin (OT) on prostaglandin F2 alpha (PGF) release from caruncular and intercaruncular endometrial tissues and prostaglandin E2 (PGE) release from intercaruncular tissue. The previously gravid uterine horn was removed on d 20 postpartum (n = 7), on d 30 postpartum (n = 7) or the uterine horn ipsilateral to the dominant follicle was removed 12-18 h after onset of first behavioral estrus postpartum (ES; n = 8). Tissues (200 mg wet wt) were cultured in Nutrient Mixture F-10 medium in a perifusion system. The medium and tissues were aerated with 95% O2: 5% CO2 and temperatures were maintained at 39 degrees C. The flow rate was 100 microliters/min and fractions were collected at 20 min intervals for 400 min. After a 2 h settling phase, the tissues were challenged with 1, 2 or 4 micrograms [Asu1,6]-OT/ml of media for 1 h. Basal release of PGE and PGF on d 20 was greater than on d 30 and at ES (P less than .02) which were similar. All doses of OT increased PGE and PGF with both remaining elevated throughout the duration of the perifusion (P less than .008). However, there were no differences among doses. Release of PGE in response to OT on d 20 and 30, was higher than at ES (P less than .008). More PGF was released in response to OT from intercaruncular than caruncular tissue on d 20 (P less than .0001) and at ES (P less than .003). Release of PGF in response to OT on d 20 was higher (P less than .0001) than on d 30 and d 30 was higher than at ES (P less than .007). Basal and OT-induced release of PGE and PGF declined as day postpartum increased. We conclude that intercaruncular tissue released more PGF than caruncular tissue and both intercaruncular and caruncular tissue responded to OT with a sustained release of prostaglandins in a non-dose-dependent manner on d 20, 30 and at ES postpartum.

Persistence of human cytomegalovirus DNA sequences without cell-virus homology in human placental explants in culture.

Using histopathology, virology, and molecular probing, we investigated the potential of human first-trimester placental tissue to support human cytomegalovirus (HCMV) infection. Histopathologic examination of infected placental explants revealed cellular changes characteristic of HCMV infection. Immunohistochemical staining of infected explants with human convalescent sera demonstrated expression of HCMV antigens within the cytotrophoblast. Dot blot hybridization of DNA extracted from infected placental tissue using cloned Hind III "W" fragment of HCMV genome indicated the presence of HCMV sequences without virus-cell homology in infected explants from one to ten days post-infection. In general, the sequences detected by the viral probes increased in abundance with time as infection continued.

Influence of follicular components on oocyte meiosis in vitro.

Oocytes and follicular components obtained from ovaries recovered from mature Hereford cows at slaughter were used to determine follicular influence on oocyte maturation. Some oocytes were fixed immediately to determine the stage of maturation. The remaining oocytes were cultured for 32 to 34 hr in various environments to determine the influences of the granulosum and follicular fluids on meiotic changes. All noncultured oocytes had dictyate nuclei except one in premetaphase. Oocytes cultured in 50 or 100% follicular fluid or in contact with stratum granulosum cells showed some meiotic inhibition both before and after germinal vesicle breakdown (GVB). The least resumption of meiosis occurred in oocytes cultured in their intact follicles.

Human cytomegalovirus infection of human placental explants in culture: histologic and immunohistochemical studies.

The induction of human cytomegalovirus infection in human first-trimester placentas was studied with a placental explant culture model. Replication and/or release of human cytomegalovirus in placental explant cultures did not occur at any time from 1 to 10 days after infection when examined by plaque assay and analyses of extracted deoxyribonucleic acids. In contrast, typical human cytomegalovirus-induced histopathologic lesions bearing human cytomegalovirus antigens were consistently localized in the trophoblastic cells covering placental villi. These data clearly demonstrate that placental cells are permissive of latent and/or abortive human cytomegalovirus infection in vitro. Our results support the hypothesis that during human cytomegalovirus infection of pregnant women, maternal viremia or intrauterine infection results in latent human cytomegalovirus infection of placental cells that may persist during the course of pregnancy.

Ultrasonic microdissection of immature intermediate human placental villi as studied by scanning electron microscopy.

The human placenta during the first 20 weeks of gestation undergoes rapid and extensive morphological changes. Near the end of this period, the most predominant type of villus present is the immature intermediate placental villus. In order to visualize this complex structure with scanning electron microscopy (SEM), we have developed a microdissection technique to expose tissue components of the placental villus while retaining its normal histological architecture. Placental villi were initially fixed in Karnovsky's fixative, buffered formalin, or 2% osmium tetroxide solution prior to exposure to connective tissue enzymes or detergents alone or in combination. Samples were dehydrated through 100% acetone and ultrasonicated at 80 kHz for 15 minutes prior to critical point drying and SEM examination. The most satisfactory microdissections were obtained by using a combined detergent/ultrasonication technique. By means of this procedure it was possible to remove the syncytiotrophoblast to expose the underlying cytotrophoblast, basal lamina and the stromal core components of the villi. The selective removal of these structures revealed the 3-dimensional relationships of the stromal channels, reticulum cells and Hofbauer cells. Of interest was the pattern of fetal capillaries coursing parallel to the long axis of each villus and terminating in a vascular knot at the tip.

The fine structure of the chinchilla placenta.

The structure of the placental labyrinth, interlobular or "coarse" syncytium, visceral (splanchnopleuric) yolk sac, giant cells and subplacenta of the chinchilla was studied with the electron microscope. The fine structure of the interhemal membrane of the placental labyrinth was found to be hemomonochorial, consisting of a single layer of syncytial trophoblast. In this respect, the placental labyrinth was similar to that of another caviomorph rodent, the guinea pig. The labyrinthine trophoblast had pinocytotic vesicles as well as larger vaculoes and multivesicular bodies. The interlobular syncytium contained granular endoplasmic reticulum, and in one case from early in gestation there were intracisternal granules in the ER. The visceral endodermal cells of the inverted yolk sac placenta had a well-developed system of apical vesicles and tubules as well as larger cytoplasmic vacuoles. Their appearance was similar to that of endodermal cells found in other rodents which are known to absorb proteins and other substances from the uterine lumen. Towards term the giant cells were often vacuolated and contained large deposits of glycogen as well as lipid droplets. The syncytial trophoblast of the subplacenta contained numerous moderately electron-dense granules which may be secretory in function; cytotrophoblastic cells lacked these granules. The subplacental syncytium often surrounded spaces or lacunae which contained an electron-dense granular material.

The fine structure of the trophospongial layer of the kangaroo rat placenta.

The trophospongial layer of the near-term kangaroo rat placenta was examined in the electron microscope. Two ultrastructurally different cellular zones were distinguishable - an inner zone adjacent to the labyrinth and a basal zone located mesometrial to the inner zone. Inner zone cells contained a well developed granular ER and Golgi apparatus as well as polymorphous membrane-limited granules. The cells also contained modest amounts of glycogen and lipid droplets. Basal zone cells were also rich in ER; some cells had dilated ER cisternae containing a highly structured material which appears as a sheet of hexagons when viewed en face. Basal zone cells may, among other things, function as glycogen storage cells, since they had large cytoplasmic accumulations of glycogen. Unlike the situation in some other rodents, maternal blood draining from the trophospongial layer was always contained in channels lined by a layer of squamous cells which, in turm, was separated from the trophospongial cells by a basal lamina. The trophospongial zones are compared with the trophospongial regions of other rodent placentas and possible functions are considered.