Biomedical subjects

F B Wooding

Publications and source records attributed to F B Wooding.

At least 19 recordsLinked to original sources

PubMed2001

Distribution of vascular endothelial growth factor (VEGF) and its binding sites at the maternal-fetal interface during gestation in pigs.

Pigs show epitheliochorial placentation, in which the maternal uterine epithelium and the fetal trophectoderm become closely apposed. There is no invasion of trophoblast into the maternal tissue, and nutrient and waste exchange take place across two epithelial layers beneath which a complex network of capillaries forms. Later in gestation, the epithelial cells become indented by blood vessels, which greatly reduces the distance for diffusion between the two circulatory systems. Vascular endothelial growth factor is a secreted homodimeric angiogenic growth factor that is involved in physiological and pathological angiogenesis. Its receptors are generally restricted to endothelial cells. Ligand binding, in situ hybridization and immunohistochemistry were carried out in pig placenta throughout gestation to investigate the possible role of vascular endothelial growth factor and its receptors in non-invasive placentation. In situ hybridization and immunohistochemistry revealed that mRNA and immunoreactivity for vascular endothelial growth factor were localized in both maternal and fetal epithelial cells at the maternal-fetal interface and over the maternal glands, although the signal was generally weaker in the maternal glands. Ligand binding was used to localize for vascular endothelial growth factor receptors; no binding was observed over the maternal glands, but very strong binding was localized to the endometrial blood vessels. At the interface between maternal and fetal tissue, a similar pattern was observed whereby the numerous small capillaries at the bases of the two apposed epithelia bound vascular endothelial growth factor specifically. It is concluded that vascular endothelial growth factor produced by the maternal and fetal epithelial layers promotes the growth of capillaries locally, which would facilitate the development of two vascular networks for the efficient transfer of nutrients and waste products.

Animals↗

PubMed2000

Separate sites and mechanisms for placental transport of calcium, iron and glucose in the equine placenta.

The placenta is the only channel for transport of nutrients to the conceptus and the fetal nutrient demands increase exponentially to term. The 9 kDa calcium binding protein (calbindin, 9CBP) and the iron binding protein uteroferrin (UF) are proving to be reliable markers for epithelia that mediate active transcellular calcium and iron transport and the glucose transporter proteins (GT1 and GT3) for glucose transport by facilitated diffusion. Light and electron microscope immunocytochemistry have been used on perfusion fixed resin embedded material to establish the distribution of 9CBP, UF, GT1 and GT3 in the equine placenta from 100 days of pregnancy to term (336 days). The equine placenta has two main structural components, flat areolae and microcotyledons. From 100 days of pregnancy to term immunoreactive 9CBP is found only in the cytoplasm of the maternal glands and the areolar trophoblast cells with none in the microcotyledons; whereas GT1 is present exclusively in the microcotyledons on the basolateral plasmalemma of both trophoblast and uterine epithelia with GT3 on the apical microvilli. The glands show neither GT1 nor GT3 expression. The areas of both areolae and microcotyledons increase enormously during gestation but there is no indication of increasing amounts of 9CBP, GT1 or GT3 protein per cell. Glucose transport through the placental cell cytoplasm is by diffusion of the free molecule, but calcium ions in transit must be sequestered in some way since the high calcium fluxes needed to support fetal bone growth in later pregnancy would be deleterious to calcium based homeostasis and cellular control systems. Electron microscope immunocytochemistry shows that 9CBP is uniformly distributed in the cytoplasm and nucleoplasm of the areolar trophoblast cells but excluded from all membrane bounded compartments such as mitochondria, Golgi saccules and pinocytotic transport vesicles. Such apical transport vesicles can be identified immunocytochemically by their content of uteroferrin, a component of the secretion from the uterine glands. It is suggested that transcellular calcium transport is therefore based on facilitated diffusion, not the vesicular method followed by the iron in the UF molecules, with 9CBP providing both transfer and sequestration functions for the transient calcium ions. These results show that the equine placenta has transport systems with restricted regional distribution similar to those recently shown for the ruminant placenta.

Acid Phosphatase↗

PubMed2000

Glycosylation in the near-term epitheliochorial placenta of the horse, donkey and camel: a comparative study of interbreeding and non-interbreeding species.

Studies from this laboratory have shown great diversity in the glycosylation of tissues comprising the interhaemal barrier of species with different placental types. This diversity may be one of the factors preventing interbreeding between species. Glycan expression within the uterine epithelium and trophoblast of the interhaemal barrier was examined to test this proposition in three species with similar diffuse, microcotyledonary, epitheliochorial allantochorionic types of placenta: the horse (Equus caballus) and donkey (Equus asinus), which can interbreed with each other, and the camel (Camelus dromedarius), which cannot interbreed with either of the other two species. A panel of 14 lectins was used and it was found that glycosylation patterns were generally similar between placental tissues of the horse and donkey, except for the expression of non-bisected complex N-glycan and some sialic acids, whereas those of the camel showed striking differences in the binding of lectins to many structures carrying terminal residues of fucose, N-acetyl galactosamine and beta-galactose, as well as to complex N-glycans and sialic acids. These results are consistent with the proposition that interbreeding species carry similar glycans in tissues forming the interhaemal barrier whereas glycodiversity is one of the factors preventing implantation and subsequent placental development in interspecific hybrids.

Animals↗

PubMed1999

A lectin binding analysis of glycosylation patterns during development of the equine placenta.

The glycosylation of the equine interhaemal barrier and areola was studied throughout the period of gestation. Placentae of 35, 37, 50, 119, 152, 200, 280 and 300 days gestation were investigated, using semithin plastic embedded sections and a panel of 15 biotinylated lectins with an avidin-peroxidase revealing system. Glycosylation of the trophoblast and maternal epithelium showed the most change during the first 50 days of gestation, being associated with the initial stages of adhesion and attachment. In the trophoblast, non-bisected tri/tetraantennary complex N-glycan was only evident after day 37 and terminal N-acetyl galactosamine, alpha2,3- and alpha2,6-linked sialic acids disappeared at the same time. The areolar trophoblast exhibited some differences from microcotyledonary areas, especially with respect to 2-deoxy, 2-acetamido alpha-galactose and tri/tetraantennary, non-bisected complex N-glycan, suggesting that the differences in function between microcotyledonary and areolar trophoblast are reflected at both the morphological and the biochemical level. Granules of the maternal uterine epithelium bound many lectins, particularly those with specificity for bisected and non-bisected bi/triantennary N-linked glycan, 2-deoxy, 2-acetamido alpha-galactosyl, beta-galactosyl and some fucosylated termini. Binding to sialic acids in alpha2,3- and alpha2,6-linkage was sparse. Maternal and fetal capillaries showed little change in glycan expression over the period studied, being rich in bisected and non-bisected bi/triantennary N-linked glycan and sialic acids, with some terminal N-acetyl galactosamine and no detectable terminal fucosyl residues.

Acetylgalactosamine↗

PubMed1998

Immunocytochemical and In situ hybridization studies of the distribution of calbindin D9k in the bovine placenta throughout pregnancy.

The fetus must transport considerable and increasing amounts of calcium across the placental trophoblast epithelium to support growth and development and bone formation. Active calcium transport across epithelia has been shown to correlate with calbindin D9k or 28k content. This study examined the distribution of calbindin D9k (9CBP) protein and mRNA during pregnancy in the bovine placenta to determine its possible role in calcium transport in this system. The immunocytochemical results show 9CBP in an increasing percentage of interplacentomal uninucleate trophoblast cells until, at term, all show a level at least eight times that of any other placental cell. There is a similar, although smaller, rise in their 9CBP mRNA content. The mature interplacentomal binucleate cell ( approximately 5% of the total) contains no 9CBP at any stage of pregnancy. In interplacentomal uterine epithelium, 9CBP protein and mRNA decrease to zero in late pregnancy but the glands maintain constant low levels throughout. In the placentome trophoblast, uninucleate cells show insignificant amounts but binucleate cells (15-20% of the total trophoblast cells) contain considerable levels of both 9CBP protein and mRNA, as do all the uninucleate uterine epithelial cells. The placentomal binucleate cells show peak values at mid-pregnancy; the placentomal uterine epithelium shows only small changes in levels in the second half of pregnancy. Increase in fetal calcium demand in the second half of pregnancy therefore correlates with a major increase in 9CBP only in the interplacentomal trophoblast, as we have also shown in the sheep and goat, indicating an important role for this region in active calcium transport by the ruminant placenta. The 9CBP is distributed uniformly in the cytosol and nucleoplasm, supporting a role in facilitated diffusion of calcium through the cell rather than a vesicular shuttle system.

Animals↗

PubMed1997

Metabolism and possible compartmentalization of inositol lipids in isolated rat-liver nuclei.

(1) The removal of the nuclear envelope from isolated rat-liver nuclei by washing with Triton X-100 (TX-100) was assessed by electron microscopy. All the envelope was removed by 0.04% (w/v) TX-100. (2) After this removal, phosphorylation of inositol lipids and diacylglycerol (DAG) from [gamma-32P]ATP still occurs, despite the near complete absence of detectable (by mass assay) DAG and PtdIns. This suggests that the majority of these two lipids in nuclei are present in the nuclear membrane, but the small amounts remaining after extraction, defined as intranuclear, are available for phosphorylation by lipid kinases (36% for DAG and 24% for PtdIns respectively, when expressed as a percentage of incorporation of intact nuclei). (3) PtdIns(4,5)P2 did not follow the same pattern as PtdIns and DAG; after removal of the nuclear membrane, 40% of the mass of this lipid was left in the nucleus. Moreover, a similar amount of PtdIns(4,5)P2 was also resistant to extraction with even higher concentrations of detergent, suggesting that PtdIns(4,5)P2 has a discrete intranuclear location, probably bound to nuclear proteins. (4) Addition of exogenous substrates, PtdIns, PtdIns(4)P and DAG, to membrane-depleted nuclei resulted in reconstitution of the majority of lipid phosphorylations from [gamma-32P]ATP (70%, 90% and 94% of intact nuclei respectively), suggesting a predominantly intranuclear location for the respective kinases. (5) Nuclei also showed phosphomonoesterase and phosphatidic acid hydrolase activity; dephosphorylation of pre-radiolabelled PtdIns(4)P, PtdIns(4,5)P2 and phosphatidic acid was observed when [gamma-32P]ATP was removed. However, some of the radioactivity was apparently resistant to these enzymes, suggesting the existence of multiple pools of these lipids. (6) Addition of excess non-radiolabelled ATP to nuclei pre-labelled with [gamma-32P]ATP resulted in an initial increase in the label in PtdIns(4,5)P2, implying a precursor-product relationship between the radiolabelled pools of PtdIns(4)P and PtdIns(4,5)P2. This was confirmed by analysis of the incorporation of 32P into the 4'-phosphate group of PtdIns(4)P and the individual 4'- and 5'-phosphate groups of PtdIns(4,5)P2. The data from these experiments also indicated that PtdIns(4,5)P2 can be produced from a pre-existing pool of PtdIns(4)P, as well as de novo from PtdIns. (7) Taken together our data suggest that isolated rat-liver nuclei have an intranuclear inositol lipid metabolism mechanism utilizing enzymes and substrates equivalent to those found in cytosol and plasma membrane, and that there may be some, but not complete, compartmentalization of the components of the nuclear inositol cycle.

Adenosine Triphosphate↗

PubMed1997

Aspects of calcium transport by the ovine placenta: studies based on the interplacentomal region of the chorion.

An in vitro technique for the measurement of calcium uptake into the maternal-facing fetal chorionic membrane (apical trophoblast) was used to study the relationship between calcium uptake and stage of pregnancy in the sheep. The effects on calcium uptake of varying calcium concentration and temperature of the incubation medium, of adding calcium channel blockers or heavy metals (lanthanum and nickel) or calcium ionophore/agonist were also studied. The data indicate a saturable calcium uptake process, plateauing after 15 min incubation. This uptake remained constant throughout the last third of gestation until a significant fall in uptake was noted during the final week prior to parturition. This uptake was not due to extracellular cellular diffusion since there was no significant uptake of tritiated inulin over the same period in each case. Calcium uptake in this system was also shown to be a temperature dependent process which was abolished at temperatures of 0-4 degrees C. A decrease in calcium concentration to 0.12 mM in the incubation medium also caused a corresponding decrease in calcium uptake to 21 per cent of control (1.2 mM). The addition of the heavy metals lanthanum and nickel also significantly reduced calcium uptake as did the calcium channel blockers verapamil, metoprolol and diltiazem. The calcium channel ionophore A23187 increased calcium uptake into the material facing chorion. Although the interplacentomal chorion may not be representative of the whole of the placental unit, it clearly contains a specific calcium uptake process under local physiological control. The blocking of calcium uptake by the specific I-type calcium channel blocker verapamil may indicate the presence of I-type channels of unusually low sensitivity since the concentration needed to block them was much higher than would be required for excitable I-type channels in isolated cells.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

PubMed1997

Quantitative analysis throughout pregnancy of intraepithelial large granular and non-granular lymphocyte distributions in the synepitheliochorial placenta of the cow.

Intraepithelial lymphocytes are a constant feature of ruminant uterine epithelium. Light microscope quantitation on semithin sections of resin embedded perfused material from pregnant and non-pregnant cows shows that although the proportion of large granular lymphocytes to non-granular lymphocytes in interplacentomal areas increases during pregnancy, the total number of lymphocytes in these areas remains at a similar level. However all types of lymphocytes are eliminated from the caruncular uterine epithelium of the cow by 28 days of pregnancy at the initiation of placentomal development. Despite the subsequent enormous growth in area of this region during pregnancy, no lymphocytes are found in the bovine placentomal cellular uterine epithelium. This pattern is similar to that in the ewe and goat although in these species the placentomal uterine epithelium is modified to maternofetal hybrid syncytial plaques. However, in the deer, a similar cellular placentomal epithelium has numerous intraepithelial lymphocytes and large granular lymphocytes are closely associated with fetomaternal hybrid trinucleate cells formed by binucleate cell migration throughout pregnancy. Cow interplacentomal large granular lymphocytes and trinucleate cells show similar apposition but the formation and fate of cow placentomal trinucleate cells does not involve lymphocytes. Since the caruncular (placentomal) area is 10-20 times that of the intercaruncular this suggests a very different function for intraepithelial lymphocytes in the deer compared with the other ruminant species.

Animals↗

PubMed1996

Phosphatidylserine-mediated adhesion of T-cells to endothelial cells.

Phosphatidylserine was exposed on the surface of human umbilical endothelial cells (ECV304) a few minutes after adding thrombin in vitro, as monitored by prothrombinase assays with and without annexin V. Jurkat T cells adhered to the thrombin-treated cells. The adhesion was inhibited by annexin V, indicating that it was mediated by exposed phosphatidylserine on the endothelial cells.

Annexin A5↗

PubMed1996

Localization and function of tyrosine-phosphorylated protein in pig oocytes.

Several proteins with phosphorylated tyrosine residues have been shown to be closely involved in the control meiotic nuclear division. We identified a 42-kD protein in pig oocytes, using a polyclonal antibody to a synthetic phosphotyrosine construct that increases significantly in amount after 12 hr of maturation culture, and is discretely localized to condensing and condensed chromosomes. However, since microinjection of the antibody into oocytes blocks spindle formation, the role of this protein appears to be at that stage rather than directly in chromosome condensation. Specificity of action of the 42-kD protein indicates that it may be a phosphorylation-dependent component necessary for successful spindle assembly.

Animals↗

PubMed1996

Calcium transport and the localisation of calbindin-D9k in the ruminant placenta during the second half of pregnancy.

In late pregnancy the sheep fetus requires 3 g of calcium per day, all of which must be transported across the trophoblast epithelium of the placenta. Such high levels of calcium transport across other epithelia are normally associated with the presence of calbindin-D9 or -28k. Our immunocytochemical results show that ovine, bovine and caprine interplacentomal trophoblast have high levels of calbindin-D9k, about eight to ten times more than in the placentomal region. The protein is detectable only in the uninucleate trophoblast cells in sheep and goat, the frequent binucleate cells show none. The calbindin-D9k is also present in the maternal glandular epithelium but not the surface epithelium of the uterus. The cellular distribution of the calbindin-D9k immunoreactivity suggests a soluble protein homogenously distributed through cytosol and nucleoplasm but absent from all organelles and intercellular spaces. In contrast, the uterine milk protein(s) are localised in Golgi cisternae and secretory vesicles in gland cells and in apical small endocytic vesicles and lysosomes in the uninucleate trophectodermal cells. The distribution of calbindin-D9k supports the concept that it mediates the high calcium flux by facilitated diffusion and not via any vesicular, membrane-bounded system.

Animals↗

PubMed1996

Functional specialization in the ruminant placenta: evidence for two populations of fetal binucleate cells of different selective synthetic capacity.

Trophoblast binucleate cells (BNC) in the ruminant placenta demonstrate a characteristic development, mature structure and migratory capacity whether situated in cotyledonary or intercotyledonary regions of the placenta. However, previous immunocytochemical studies demonstrated clear differences in gene expression in granule contents of BNC according to their anatomical location with some proteins being expressed in all BNC (e.g. ovine placental lactogen) whereas others were unique to a particular origin (e.g. SBU3 antigen in cotyledonary BNC only). We have used enriched preparations of binucleate cells and showed differences in steroid metabolic capacity in vitro which is more related to their species origin (sheep or goat) than to their anatomical location. The predominant product from [3H]pregnenolone is progesterone (sheep) and 5 beta-pregnane-3 alpha, 20 alpha-diol (goat) and the amount formed (corrected for the number of BNC) is similar irrespective of whether BNC were derived from the cotyledonary or intercotyledonary regions. These studies indicate specific forms of regional functional specialization of BNC and emphasize their multifunctional role in the ruminant placenta.

Animals↗

PubMed1996

Implantation and early placentation in the one-humped camel (Camelus dromedarius).

The ovaries and uteri were removed from four pregnant camels on days 14, 25, 35 and 56 after ovulation. The day 14 and 25 uteri were perfuse-fixed with 3 per cent glutaraldehyde: 3 per cent paraformaldehyde whereas the day 35 and 56 specimens were opened dorsally for photography before biopsies of allantochorion attached to endometrium were taken and fixed in either 3 per cent glutaraldehyde: 3 per cent paraformaldehyde or Bouin's fluid. Samples of each uterus were then processed and sectioned for light and transmission electron microscopy. By day 14 the majority of the trophoblast had become closely apposed to the luminal epithelium of the endometrium to form the start of an epitheliochorial placenta with microvillar interdigitation initiated in some places. By day 25 a well-developed microvillar junction had formed between the fetal and maternal tissues. The fetus was situated in the middle of the left uterine horn in the day 35 and 56 specimens and, histologically, large multinucleate giant trophoblast cells had developed at frequent but irregular intervals along the, otherwise single-cell, trophoblast. These giant cells were often situated over the mouth of an endometrial gland but their actual function in pregnancy is not yet known.

Animals↗

PubMed1995

Differences in E-selectin expression and leucocyte infiltration induced by inflammatory agents in a novel subcutaneous sponge matrix model.

We have developed a novel subcutaneous sponge matrix model in major histocompatibility complex (MHC) homozygous SLAb/b inbred pigs to study lymphocyte-endothelial cell interactions during inflammation. Polyether sponges were implanted subcutaneously and left for 12 days before injection of proinflammatory agonists. Implanted sponges became highly vascularized and showed markedly increased uptake of i.v.-injected 51Cr-labelled lymphocytes 5 hr after injection of tumour necrosis factor-alpha (TNF-alpha) (3000 U) or phytohaemagglutinin (PHA) (37 micrograms). Lower levels of traffic were seen in sponges 5 hr after injection with interleukin-1 alpha (IL-1 alpha) (3000 U) and no significant traffic occurred in sponges injected with phorbol 12-myristate 13-acetate (PMA) (15 ng) at 5 hr or PHA at 24 hr (compared to sponges injected with medium alone). Electron microscopy of control sponges revealed low numbers of infiltrating leucocytes and relatively 'flat' endothelium. Many more infiltrating leucocytes were present in PHA-injected sponges. However, no ultrastructural evidence was seen of any significant difference between control and activated endothelium. Immunocytochemistry of frozen sections from sponges showed that E-selectin expression was up-regulated markedly by TNF-alpha and PHA at 5 hr, only moderately by IL-1 alpha at 5 hr, and not at all by PMA at 5 hr. By 24 hr in PHA-injected sponges E-selectin expression had fallen markedly from the level seen at 5 hr. Flow cytometric analysis of cellular infiltrates dispersed from sponges injected with TNF-alpha, PHA, IL-1 alpha or medium alone, revealed differences in lymphocyte subset populations. The infiltrate in sponges injected with TNF-alpha 5 hr before removal was dominated by high numbers of CD2+ lymphocytes, whereas the infiltrate induced by PHA showed relatively higher levels of CD2- CD4-CD8- gamma delta T-cell receptor+ (TCR+) T cells revealed by population-specific monoclonal antibodies (mAb). This model, which permits harvesting of leucocytes and endothelial cells for manipulation in vitro, will be useful for the study of leucocyte-endothelial cell interactions in subacute and chronic inflammation.

Animals↗

PubMed1995

Quantitative analysis of intraepithelial large granular lymphocyte distribution and maternofetal cellular interactions in the synepitheliochorial placenta of the deer.

Intraepithelial lymphocytes are a constant feature of ruminant uterine epithelium in nonpregnant animals. Quantitation in 6 species of deer shows that the proportion of large granular lymphocytes (LGL) in this population increases markedly and continuously from the earliest implantation to the latest (midpregnant) stage examined. The size of individual granules also increases. Fetal trophectodermal binucleate cells are also present from the earliest stage in all deer examined and follow a usual ruminant life cycle: migrating and fusing with uterine epithelial cells to form trinucleate cells before releasing their granules to the maternal compartment and finally degenerating and being reabsorbed by the trophectodermal uninucleate cells. Deer LGLs were usually closely associated with the degranulating fetomaternal hybrid trinucleate cells but showed no ultrastructural changes themselves. This association indicates a dynamic interaction between deer LGL and trinucleate cells which could serve as one of the factors restricting the extent of trinucleate cell progression to a continuous syncytium as found in sheep and goats.

Animals↗

PubMed1994

Membrane dynamics during migration of placental cells through trophectodermal tight junctions in sheep and goats.

Binucleate cells in ruminant trophectodermal epithelium are unique in that they form part of the tight junction as they migrate across it, maintaining the ionic barrier seal to the internal milieu of the fetus. Such participation imposes considerable constraints on the cell migration because membrane cannot flow through a tight junction. We report quantitative ultrastructural immunocytochemical evidence for vesicle membrane insertion into the binucleate cell plasmalemma which allows the cells to form a pseudopodium past the tight junction. This pseudopodium increases continuously in area by vesicle insertion and develops a close apposition to the plasmalemma of the fetomaternal syncytium which constitutes the fetomaternal boundary in the placenta of the sheep and goat. Eventually the apposed membranes of the binucleate cell pseudopodium and the syncytium fuse by vesiculation and the cytoplasm and nuclei of the binucleate cell merge into the fetomaternal syncytium. The binucleate cell plasmalemma remaining on the trophectodermal side of the tight junction is blebbed off into, and phagocytosed by, the uninucleate trophectodermal cells between which the binucleate cell passed. This process permits the delivery of the binucleate cell granules to the maternal side of the placenta but none of the fetal molecules expressed on the plasma membrane of the binucleate cells are exposed to potential maternal immunological rejection.

Animals↗

PubMed1993

Giant and binucleate trophoblast cells of mammals.

The cellular origin, structure, and function of trophoblastic giant cells (GC) and binucleate cells (BNC) are reviewed. Mammals in which these cells have received the greatest attention include rodents, rabbits, and humans (GCs), and ruminants and equids (BNCs). In almost all cases these cells arise from the cytotrophoblast. All are large cells and contain either two diploid nuclei (BNCs), multiple nuclei (human placental bed GCs), or single nuclei with amplified DNA content (rodent and rabbit GCs). Giant and binucleate cells typically exhibit the capacity for migration or invasion, although the degree of migratory activity varies between species. While most end up within, or at the interface with, endometrial tissue, in some instances the GCs or BNCs contribute directly to the interhemal membrane of the placenta. Hormone production is a property which most GC-BNC populations have in common. Lactogen or gonadotropin has been documented in almost all cells of this type examined to date, and in some animals they are also steroidogenic (e.g., rats and sheep). In spite of some common features, both structural and functional differences remain and it is suggested that use of terms such as mononuclear giant cells, multinucleate giant cells, and binucleate cells be continued rather than assuming that these cells are all members of a single trophoblastic subtype.

Animals↗

PubMed1993

Cellular dynamics of growth in sheep and goat synepitheliochorial placentomes: an autoradiographic study.

This paper demonstrates that in sheep and goats the two definitive fetomaternal interface layers are developmentally related. The fetal trophectoderm consists of binucleate and uninucleate cells. The apical microvilli of the trophectoderm interdigitate with a layer consisting of syncytial plaques of limited area bounding the maternal connective tissue. Our previous histological ultrastructural and immunocytochemical work has indicated that throughout pregnancy the fetal binucleate cells migrate to and fuse with the uterine epithelium or its derivatives to form these syncytial plaques which constitute a persistent fetomaternal tissue unique to ruminants. This quantitative autoradiographic study of thymidine incorporation into sheep and goat placentas confirms the central role of the binucleate cell in placental growth, demonstrates that throughout pregnancy all binucleate cells migrate and indicates that most of the nuclei of the syncytial plaques, which appear to have a limited lifespan, derive from binucleate cell fusion.

Animals↗