Studies of the dynamics of nuclear clustering in human syncytiotrophoblast.

Syncytial nuclear aggregates (SNAs), clusters of nuclei in the syncytiotrophoblast of the human placenta, are increased as gestation advances and in pregnancy pathologies. The origins of increased SNAs are unclear; however, a better appreciation of the mechanism may give insight into placental ageing and factors underpinning dysfunction. We developed three models to investigate whether SNA formation results from a dynamic process of nuclear movement and to generate alternative hypotheses. SNA count and size were measured in placental explants cultured over 16 days and particles released into culture medium were quantified. Primary trophoblasts were cultured for 6 days. Explants and trophoblasts were cultured with and without cytoskeletal inhibitors. An in silico model was developed to examine the effects of modulating nuclear behaviour on clustering. In explants, neither median SNA number (108 SNA/mm(2) villous area) nor size (283 µm(2)) changed over time. Subcellular particles from conditioned culture medium showed a wide range of sizes that overlapped with those of SNAs. Nuclei in primary trophoblasts did not change position relative to other nuclei; apparent movement was associated with positional changes of the syncytial cell membrane. In both models, SNAs and nuclear clusters were stable despite pharmacological disruption of cytoskeletal activity. In silico, increased nuclear movement, adhesiveness and sites of cytotrophoblast fusion were related to nuclear clustering. The prominence of SNAs in pregnancy disorders may not result from an active process involving cytoskeleton-mediated rearrangement of syncytial nuclei. Further insights into the mechanism(s) of SNA formation will aid understanding of their increased presence in pregnancy pathologies.

Tracking nutrient transfer at the human maternofetal interface from 4 weeks to term.

INTRODUCTION: In this study we have tracked glycogen and glycoprotein flux associated with nutrient uptake into trophoblast in early deciduochorial and later haemochorial placenta. METHODS: α-amylase, glycogen synthase and glycogen phosphorylase were immunohistochemically localised in 6-14 week and term placenta and first trimester decidua. Placentae of 4-18 weeks' gestation and term were also stained with 22 biotinylated lectins. RESULTS: Histochemical data were consistent with glycogenolysis in decidual gland epithelium and placental cyto- and syncytiotrophoblast; α-amylase was present in decidual secretions but absent in placenta. Glycogen and glycogen synthase were both apparent in villous cytotrophoblast cells and columns. Profound changes were observed in placental glycosylation during gestation. Syncytial microvilli were richly glycosylated as were first trimester vacuoles but, by term, syncytiotrophoblast showed little lectin binding except in microvillous and basal membranes. Cytotrophoblast Golgi bodies were active in the first trimester; at term the cells were generally more glycosylated than syncytiotrophoblast. DISCUSSION: We deduce that decidual cell glycogen is broken down for transport into the placenta where the products may be reassembled into glycogen or used for metabolic processes. First trimester histiotrophe is internalised by syncytiotrophoblast, then broken down in apical vacuoles containing lysosomal markers. This process declines after haemotrophic nutrition commences. Transition from histiotrophic to haemotrophic nutrition involves reduced amounts of uterine secretory derivatives reaching the placenta, and reduction in internalisation of glycoprotein by syncytiotrophoblast, presumably reflecting the shift to low molecular weight nutrients. Glycogen accumulates in cytotrophoblast from early pregnancy and is mobilised for utilisation by fetoplacental tissues.

Immunocytochemistry of the placentas of giraffe (Giraffa cameleopardalis giraffa) and okapi (Okapi johnstoni): comparison with other ruminants.

INTRODUCTION: The trophoblast binucleate cell [BNC] is central to the structure and function of all ruminant placentas so far investigated. The Giraffidae are considered to form a separate family within the ruminant suborder. METHODS: The structure and function of two [mid and late pregnant] giraffe placentas and two term okapi placentas have been investigated immunocytochemically. RESULTS: Their major characteristics: polycotyledonary epitheliochorial structure, sequential glucose transport using two transporter isoforms, expression of water transporters in the interplacentomal [IP] and placentomal [P] trophoblast and restriction of calcium transport to the IP trophoblast are similar to those of the ruminant family Bovidae. . Giraffe and okapi also show characteristic ruminant trophoblast binucleate cells (BNC) which migrate and fuse with individual uterine epithelial cells as in the cow. However, there are many fewer BNC, of limited distribution, when compared with other ruminants so far investigated. The giraffe and okapi BNC also show a different range of proteins, Pregnancy Associated Glycoproteins (PAGs) and glycans which clearly distinguish the Giraffidae from the Bovidae. CONCLUSIONS: The results support a separate giraffid family derived from a common ancestry, possessing subpopulations of BNC with potentially different functions.

Trophoblast specialisations during pregnancy in the tammar wallaby, Macropus eugenii: a morphological and lectin histochemical study.

INTRODUCTION: The tammar wallaby has a short gestation (26.5 days) and vascular modifications to expedite transport during that brief pregnancy. Here we examine trophoblast structural attributes that would facilitate materno-fetal exchange. MATERIALS AND METHODS: Four specimens of Macropus eugenii between days 23 and 26 gestation were examined using electron microscopy and 24 lectins to characterise glycosylated secretions and their internalisation. RESULTS: Two trophoblast phenotypes were found, flattened cells generally in contact with the underlying uterine epithelium and giant cells associated with histiotrophe. The latter appeared to penetrate uterine clefts, occasionally detach and become necrotic. Lectin histochemistry and ultrastructure indicated the presence of many lysosomes and residual bodies especially in trophoblast giant cells; these contained glycans, mainly apically, which were also detected in secretions and cell debris. Trophoblast basal membranes bore extensive filopodia. Giant cells were less common in vascular trilaminar areas and here the trophoblast barrier became thinner near term. DISCUSSION: Loss of Maackia amurensis agglutinin binding suggested cleavage of terminal sialic acid residues as an early post-internalisation event in the trophoblast. Lectin staining indicated degradation occurred in an apical-basal direction, and the heavily glycosylated basal membrane appeared specialised for transport out of the cell. CONCLUSION: Granules seen ultrastructurally and histochemically, particularly in giant trophoblast cells of the bilaminar area, suggest that internalised histiotrophe is broken down here and nutrients transferred to the embryo via the specialised basal plasma membrane. The trilaminar vascular area contained mostly flattened trophoblast cells, supporting the suggestion that gaseous exchange is its primary function.

Analysis of syncytial nuclear aggregates in preeclampsia shows increased sectioning artefacts and decreased inter-villous bridges compared to healthy placentas.

Syncytial nuclear aggregates (SNAs) are increased in pregnancy complications and include 'true' syncytial knots and inter-villous bridges. Apparent nuclear overlay caused by sectioning artefacts are frequently counted from single sections. Haematoxylin and eosin stained serial sections were assessed for frequency of SNA subtypes in placentas from normal, preeclamptic and fetal growth restricted (FGR) pregnancies. There were more sectioning artefacts and syncytial knots and fewer bridges in samples from preeclampsia compared to controls. There were no significant differences between FGR and control samples. This suggests the villous tree in preeclampsia has less inherent structural support and trophoblast cell dynamics are different.

The fetomaternal interface shows vascular hypoglycosylation in the tammar wallaby Macropus eugenii: comparison with a range of non-mammalian and eutherian placentae.

INTRODUCTION: Blood vessel glycosylation at the fetomaternal interface of four near-term specimens of tammar wallaby, Macropus eugenii, has been examined at days 23-26 of the 26.5 day pregnancy and compared with that of other species. METHODS: A panel of 23 lectins was used to compare vasculature in tammar with non-mammalian (shark, skink) and eutherian species at early and late gestation (camel, horse and alpaca), and term/near-term (cat, lion, dog, mink and elephant). RESULTS: Strikingly low levels of all the glycans tested, apart from sialic acids, were found in capillary endothelium of both the trilaminar omphalopleure and underlying surface endometrium of the tammar, though deeper endometrial vessels showed normally high levels of glycosylation. Only maternal vasculature of the mink placenta showed a comparable lack of expression. DISCUSSION: One reason for a reduced endothelial glycocalyx may be to facilitate diffusion of gases and nutrients as the tammar trophoblast lacks the indentation by overlying vessels that is seen in the other near-term placentae. Early epitheliochorial placentae of other species with equal diffusion distances to the tammar, showed normal vascular glycosylation. However, their pregnancies are much longer. CONCLUSION: The hypoglycosylation of tammar vessels at the fetomaternal interface may allow continued transfer of nutrients and gaseous exchange during the extremely rapid period of organogenesis which occurs during the short 26.5 day pregnancy of this marsupial. Given the short gestation period of the tammar, we suggest that a thinner endothelial glycocalyx has evolved to facilitate diffusion of gases and nutrients between the maternal and fetal compartments.

A new form of rodent placentation in the relict species, Laonastes aenigmamus (Rodentia: Diatomyidae).

INTRODUCTION: The Laotian rock rat is a relict species in a sister group relationship to hystricognath rodents (Hystricognathi). We asked whether there were similarities in placentation that might reflect this relationship or differences that might cast light on the evolution of Hystricognathi. METHODS: We examined the reproductive tract of nonpregnant (n = 5), early (n = 3) and mid to late gestation (n = 2) females. Selected characters were mapped to a phylogenetic tree to examine their evolution in rodents. RESULTS: The chorionic placenta was discoid and labyrinthine with a spongy zone but without internal lobes. The interhemal region was hemodichorial with syncytiotrophoblast lining maternal blood spaces and an inner layer of vacuolated cytotrophoblast. There was no subplacenta. The yolk sac was well developed with a villous portion that faced the placental disk but no fibrovascular ring. There was a single fetus that very likely would be precocial at birth. DISCUSSION: A lobulated labyrinth and the presence of a subplacenta and a fibrovascular ring emerged as synapomorphies for Hystricognathi. Laonastes, Ctenodactylus and stem Hystricognathi all had precocial young, whereas altriciality was the plesiomorphic condition for rodents. A hemomonochorial interhemal region was plesiomorphic for rodents and Hystricognathi, and the hemodichorial condition found in Laonastes, and possibly in Ctenodactylus, was unlike that of any rodent studied to date. CONCLUSION: Similar to Hystricognathi, Laonastes bears precocial young, but this species lacks placental adaptations such as the subplacenta, suggesting they were evolved subsequent to a change in reproductive strategy in the common ancestor of Laonastes and Hystricognathi.

Syncytial nuclear aggregates in normal placenta show increased nuclear condensation, but apoptosis and cytoskeletal redistribution are uncommon.

INTRODUCTION: Syncytial nuclear aggregates (SNAs) are increased in pregnancy complications; however, little is known about their origin or function. This study aimed to characterise SNAs in more detail than has been reported previously. METHODS: Immunohistochemistry and morphological examination at the light and ultrastructural level were used to determine the nature and structure of SNAs. RESULTS: SNAs comprising bridges and syncytial knots had similar frequency with 974 per mm3 of villous tissue (IQR 717-1193) and 833 per mm3 (IQR 766-1190), respectively while there were approximately four times as many sectioning artefacts than knots and bridges combined. SNAs had increased proportions of condensed nuclei compared to the remaining syncytiotrophoblast (33.3% vs. 8.9%) and decreased proportions of euchromatic nuclei (0.0% vs. 16.2%), as assessed by examination of an electron micrograph archive. SNAs showed little evidence of apoptosis, with weak positivity for the apoptosis markers M30-neoepitope at 16.6% and TUNEL at 10.0%; strong staining was rarely seen for either marker. Immunofluorescence demonstrated rare association of actin (α, ß or γ) with SNAs, whereas tubulin was in close proximity to SNAs and cytokeratin was seen within and surrounding SNAs. DISCUSSION: M30-positive SNAs traced through serial sections were significantly more likely to be syncytial knots or sectioning artefacts than bridges. Nuclei within SNAs showed signs consistent with degeneration; however, this is unlikely to be an apoptotic process. There are few changes in configuration of cytoskeletal proteins around SNAs. CONCLUSIONS: These data suggest that the biogenesis and functional significance of SNAs still require resolution.

IFPA Meeting 2011 workshop report III: Placental immunology; epigenetic and microRNA-dependent gene regulation; comparative placentation; trophoblast differentiation; stem cells.

Workshops are an important part of the IFPA annual meeting as they allow for discussion of specialised topics. At IFPA meeting 2011 there were twelve themed workshops, five of which are summarized in this report. These workshops related to various aspects of placental biology: 1) immunology; 2) epigenetics; 3) comparative placentation; 4) trophoblast differentiation; 5) stem cells.

First trimester histiotrophe shows altered sialylation compared with secretory phase glycoconjugates in human endometrium.

OBJECTIVES: Histiotrophe is now recognized as being an important feature of early human pregnancy, providing nutrients and growth factors to the developing embryo. Our aim was to examine the glycan composition of histiotrophe from first trimester decidua and to compare it with secretions present in endometrial glands from the late secretory phase of the menstrual cycle. STUDY DESIGN: Twenty samples of decidua from pregnancies between 8 and 11 weeks were processed into epoxy resin and sections stained with a panel of 22 lectins, together with six late secretory phase endometrial biopsies. MAIN OUTCOME MEASURES: Specimens were analysed using a semi-quantitative ranking system and the density of lectin binding to the glandular secretions and the epithelium assessed. RESULTS: With the onset of pregnancy, beta-galactose, alpha-N-acetyl galactosamine and N-Acetyl lactosamine bound by Arachis hypogaea, Glycine max, Helix pomatia and Erythrina crystagalli agglutinins appeared in terminal positions on oligosaccharide chains, suggesting loss of the capping sialic acid residues present in the non-pregnant state. CONCLUSIONS: Suppression of terminal sialylation is evident during early pregnancy, suggesting that modifications to endometrial glandular activity occur in response to placental signals. The changes may facilitate absorption of histiotrophe by the trophoblast and enhance availability of substrates for degradation.

Characterisation of Hofbauer cells in first and second trimester placenta: incidence, phenotype, survival in vitro and motility.

Macrophages, known as Hofbauer cells, are most abundant in placental villous stroma in the first and second trimesters. Their functions are not well defined. We have used a combination of in situ and in vitro methods to characterise these cells. Lectin histochemistry and immunohistochemistry were used to identify macrophages in situ. The lectin from Maclura pomifera (MPA) was found to mark cells bearing the CD68 antigen with optimal specificity and selectivity. MPA staining was used to show that they increase in number from mid first to mid second trimester, becoming much less abundant at term. The cells are absent from mesenchymal villi, being associated primarily with villous stroma containing the prominent interstitial channels characteristic of immature intermediate villi. A mixed stromal cell isolate was studied in monolayer culture, including the use of time-lapse microscopy. Cells from first or second trimester tissue contained a subpopulation of about 14-17% of cells that exhibited a macrophage-like morphology and expressed CD68 as well as MPA-binding glycans. These cells were short-lived in monoculture, but could persist in vitro in association with a fibroblast layer for several weeks. They could switch rapidly from a macrophage-like to a fibroblastic morphology, were highly motile and associated in clusters that rapidly formed and dissipated over periods of a few hours. These data suggest that Hofbauer cells play a role in the maturation of mesenchymal into immature intermediate-type stroma. They may be important in the excavation of stromal channels. Their prolonged viability in mixed cultures suggests a paracrine relationship with resident fibroblasts. Their location and migratory behaviour predict an ability to move rapidly around the villous stroma, perhaps within the channel system, and to make transient contacts both with other macrophages and stromal cells.

Dexamethasone stimulates placental system A transport and trophoblast differentiation in term villous explants.

Synthetic glucocorticoids (GCs) are given to women with threatened preterm labour but their administration has been linked to reduced infant birthweight. The underlying mechanisms are unknown, but impaired placental development and/or function has been implicated. The activity of the system A amino acid transporter is decreased in placentas from pregnancies complicated by fetal growth restriction. Whether GCs adversely affect placental amino acid transport is unknown. The objective of this study was to determine the regulatory effects of GCs on system A transport using a human in vitro placental explant model. Term explants (n=7) were treated with dexamethasone (DEX 10(-8)M or 10(-6)M) or vehicle for 48 h. System A activity was measured by the uptake of (14)C-N-methylated aminoisobutyric acid by explants. Explants were also processed for electron microscopy (EM), immunohistochemistry, and qRT-PCR. Lactate dehydrogenase (LDH), human chorionic gonadotropin (hCG) and human placental lactogen (hPL) release into the culture medium was measured. DEX (10(-6)M) stimulated system A activity compared to vehicle (p<0.05). System A transporter proteins were localized to the newly regenerating syncytiotrophoblast layer, but mRNA levels were unchanged with DEX treatment. DEX did not adversely affect explant viability as determined by reduced LDH release (p<0.05). DEX treatment was associated with morphological (accelerated apical microvilli formation, nuclear maturation, and increased cell organelle number) and functional (elevated hCG secretion, increased 11beta-HSD2 mRNA expression and reduced cytotrophoblast proliferation (p<0.05 for all)) markers of syncytiotrophoblast differentiation. These findings suggest that DEX stimulates system A activity and promotes syncytiotrophoblast differentiation and maturation.

Isolation of plasma membrane vesicles from mouse placenta at term and measurement of system A and system beta amino acid transporter activity.

Placental amino acid transport is essential for optimal fetal growth and development, with a reduced fetal provision of amino acids being implicated as a potential cause of fetal growth restriction (FGR). Understanding placental insufficiency related FGR has been aided by the development of mouse models that have features of the human disease. However, to take maximal advantage of these, methods are required to study placental function in the mouse. Here, we report a method to isolate plasma membrane vesicles from mouse placenta near-term and have used these to investigate two amino acid transporters, systems A and beta, the activities of which are reduced in human placental microvillous plasma membrane (MVM) vesicles from FGR pregnancies. Plasma membrane vesicles were isolated at embryonic day 18 by a protocol involving homogenisation, MgCl(2) precipitation and centrifugation. Vesicles were enriched 11.3+/-0.5-fold in alkaline phosphatase activity as compared to initial homogenate, with minimal intracellular organelle contamination as judged by marker analyses. Cytochemistry revealed alkaline phosphatase was localised between trophoblast layers I and II, with intense reaction product deposited on the maternal-facing plasma membrane of layer II, suggesting that vesicles were derived from this trophoblast membrane. System A and system beta activity in mouse placental vesicles, measured as Na(+)-dependent uptake of (14)C-methylaminoisobutyric acid (MeAIB) and (3)H-taurine respectively confirmed localisation of these transporters to the maternal-facing plasma membrane of layer II. Comparison to human placental MVM showed that system A activity was comparable at initial rate between species whilst system beta activity was significantly lower in mouse. This mirrored the lower expression of TAUT observed in mouse placental vesicles. We conclude that syncytiotrophoblast layer II-derived plasma membrane vesicles can be isolated and used to examine transporter function.

Altered glycosylation in peri-implantation phase endometrium in women with stages III and IV endometriosis.

BACKGROUND: Endometriosis is a common cause of infertility and pelvic pain. Lectin histochemistry has shown that glycan expression is a sensitive marker of differentiation in the normal endometrium. Endometrial biopsies were taken during the implantation window from women with subfertility and advanced (stage III and IV) endometriosis to evaluate specific glycans bound by lectins from Dolichos biflorus agglutinin (DBA) and Vicia villosa agglutinin (VVA), which detect related but distinct glycan sequences regulated by progesterone action. METHODS: Endometrial tissue from 12 women with subfertility and advanced endometriosis and 11 healthy controls were taken on days 19-24 of the menstrual cycle and processed into either epoxy resin or paraffin wax. Lectin histochemistry was analysed using light microscopy to quantify the amount of glandular reaction product. RESULTS: There was a significant (P = 0.011) reduction in DBA binding to endometrium from patients with endometriosis compared with controls, which was not seen with VVA (P = 0.135). Three stage IV biopsies and one stage III biopsy completely failed to bind DBA and, of these, three showed moderate glandular binding of VVA. DBA and VVA binding differed significantly (P= 0.0039) in the endometriosis specimens whereas in controls no significant difference was detected (P = 0.812). CONCLUSION: Secretory phase glycosylation in women with advanced endometriosis differs from that in healthy women with a reduction in fucosylated N-acetylgalactosamine sequences bound by DBA. Shorter VVA-binding glycans are not significantly affected. In addition to indicating abnormalities of epithelial differentiation, these findings may be directly relevant to implantation failure, as blastocyst attachment requires a critical interaction with the epithelial glycocalyx.

The glycosylation pattern of secretory granules in binucleate trophoblast cells is highly conserved in ruminants.

The binucleate trophoblast cells (BNCs) in the ruminant placenta are a unique feature of this taxon. These cells produce several secretory proteins and transfer these across the fetomaternal barrier into the dam. We used lectin histochemistry with a panel of 24 lectins to characterise the glycosylation pattern of BNC secretory granules in a variety of ruminants. Seven species out of three ruminant families were thus investigated: greater malayan chevrotain (Tragulidae); fallow deer, red deer, chinese water deer (Cervidae); and domestic goat, springbok, impala (Bovidae). BNC granules in all species studied strongly expressed tri-/tetraantennary complex N-glycans and bisecting N-acetylglucosamine [GlcNAc] as shown by binding of leuco- and erythroagglutins of Phaseolus vulgaris respectively. The presence of terminal N-acetylgalactosamine [GalNAc]) in BNC granules is shown by intense staining with lectins from Dolichos biflorus, Vicia villosa and Wisteria floribunda. Terminal galactose or GalNAc was also present, bound by Glycine max agglutinin. Treatment of slides with neuraminidase strongly intensified staining of Erythrina cristagalli lectin (ECA) to terminal lactosamine in all species studied; this was otherwise absent except in goat. Sambucus nigra-1 lectin bound to BNC granules in all species except in Impala, indicating the presence of abundant alpha2,6 linked sialic acid. These results indicate that these unusual highly branched glycans, with bisecting GlcNAc and terminal GalNAc are a general feature of BNC granules in Ruminants, including the most basal Tragulid branch. It therefore appears that the specific glycosylation pattern of BNC granules evolved early in ruminant phylogenesis, together with the appearance of BNC. The conserved glycan structure in BNC secretory granules indicates that this pattern of glycosylation is likely to be of considerable functional importance for the secretory glycoproteins of ruminant BNC.

Placentation in the Egyptian slit-faced bat Nycteris thebaica (Chiroptera: Nycteridae).

Bats are a highly successful, widely distributed group, with considerable variation in placental structure. The Egyptian slit-faced bat Nycteris thebaica is a member of one of the few families with previously undescribed placentation. It was found that, although the interhemal type of the Nycteris placenta is endotheliochorial with a single layer of cytotrophoblast, the arborizing pattern of the maternal vessels and especially the extraordinary major placental artery differs from the placenta of the emballonurid bats to which this family is considered to be most closely related. The major placental artery providing maternal blood to the vessels of the placental disk has a highly glycosylated matrix surrounded by two-layered folds of trophoblast, forming an apparently rigid structure of unique morphology. The yolk sac is collapsed, with hypertrophied endodermal and mesothelial cells similar to many other bat species. The paraplacenta is extensive with abundant fetal vessels underlying cytotrophoblast and syncytial trophoblast layers, fronting on an endometrium that largely lacks uterine epithelial cells but has large decidual cells and is poorly vascularized. The placenta of Nycteris lacks a hemophagous region, unlike the emballonurid bats Taphozous and Saccopteryx. Although the latter two species have similar placentas, the placental structure of Nycteris does little to relate it to the other family within the Emballonuroidea. Shared and divergent reproductive characters are discussed in relationship to bat phylogenetic relationships.

Connexin expression pattern in the endometrium of baboons is influenced by hormonal changes and the presence of endometriotic lesions.

Experimentally induced endometriosis in baboons serves as an elegant model to discriminate between endometrial genes which are primarily associated with normal endometrial function and those that are changed by the presence of endometriotic lesions. Since connexin genes are characteristic of the hormonally regulated differentiation of the endometrium, we have examined connexin expression in baboon endometrium to delineate if they are altered in response to the presence of endometriotic lesions. Connexin expression in the endometrium of cycling baboons is similar to that of the human endometrium with Connexin(Cx)43 being primarily seen in the stromal compartment and Cx26 and Cx32 being present predominantly in the epithelium. Although Cx32 is up-regulated during the secretory phase, Cx26 and Cx43 are down-regulated. In the baboon model of induced endometriosis a change in connexin pattern was evident in the presence of endometriotic lesions. In the secretory phase, Cx26 and Cx32 are no longer present in the epithelium but Cx26 is now observed primarily in the stromal cells. Infusion of chorionic gonadotrophin in a manner that mimics blastocyst transit in utero failed to rescue the aberrant stromal expression of Cx26 that is associated with the presence of endometriotic lesions suggesting an impairment of the implantation process. The altered connexin pattern coupled with a loss of the channel protein in the epithelium and a gain of Cx26 in the stromal compartment suggests that the presence of lesions changes the uterine environment and thereby the differentiation programme. This aberrant expression of connexins may be an additional factor that contributes to endometriosis-associated infertility.

Placentation in the Hottentot golden mole, Amblysomus hottentotus (Afrosoricida: Chrysochloridae).

The placentation of the Hottentot golden mole (Amblysomus hottentotus) has been examined using light and electron microscopy and lectin histochemistry of nine specimens at both mid and late gestation. The placentae were lobulated towards the allantoic surface and the lobules contained roughly parallel arrays of labyrinthine structures converging on a central spongy zone. At mid gestation, the arrays were composed of an inner cellular and outer syncytial trophoblast layer, the inner layer enclosing scant connective tissue and fetal capillaries. Maternal blood spaces coursed through the outer trophoblast and were lined by trophoblastic microvilli; the blood spaces were narrow in mid gestation but enlarged near term, while the inner trophoblast layer became thinner and seemed to be syncytial. These features were confirmed by transmission electron microscopy. The microvillous surfaces and dispersed cytoplasmic particles were heavily glycosylated, as shown by lectin histochemistry, and exhibited changes with maturation, particularly a loss in N-acetyl glucosamine oligomers bound by Phytolacca americana lectin on the microvilli lining the maternal blood spaces and outer trophoblast particles. A substantial yolk sac was present both in mid and late gestation stages. It was clearly unattached to the uterus in the later stages. These morphological features are discussed in relation to the phylogenetic position of Amblysomus with respect to other members of Afrosoricida and Afrotheria.

Adhesion molecules in human trophoblast - a review. II. extravillous trophoblast.

At the tips of anchoring villi, cytotrophoblast (CTB) proliferation leads to a process of multilayering in which cells lose their attachment to the villous basement membrane and develop into columns, within which they adhere to one another using desmosomes, with associated intermediate filament bundles. Non-desmosomal cadherins, tight junction proteins and other adhesion molecules are also present, suggesting that actin-associated adhesions contribute to placental anchorage. In the distal columns, cell-cell interactions diminish, cells upregulate beta1 integrins and bind to a provisional fibrinoid extracellular matrix, eventually detaching to migrate into the decidual stroma and myometrium, where interstitial and endovascular extravillous trophoblast (EVT) populations show distinct repertoires of adhesion molecules.

Adhesion molecules in human trophoblast - a review. I. Villous trophoblast.

In the placental villus, cells attach to basement membrane via integrin alpha6beta4 and adhere both laterally and apically to their neighbours. The most prominent adhesive specialisation seen using the electron microscope is the desmosome, which connects cytotrophoblast cells (CTB) laterally and also contributes to the attachment of CTB to the overlying syncytium. However, numerous cadherins and other junctional proteins are also present in the corresponding plasma membrane domains, indicating a multiplicity of adhesive interactions. Integrins, tight junction components and cadherins are all found in the syncytial microvillous membrane, perhaps reflecting its ability to form intersyncytial bridges. There is a wide gulf to be filled between molecular anatomy and functional studies, with much to be learned about the role of adhesion molecules in regulating villous epithelial integrity, homeostasis and growth.