Early studies of placental ultrastructure by electron microscopy.

BACKGROUND: Transmission electron microscopy (TEM) was first applied to study placental ultrastructure in the 1950's. We review those early studies and mention the scientists that employed or encouraged the use of TEM. FINDINGS: Among the pioneers Edward W. Dempsey was a key figure who attracted many other scientists to Washington University in St. Louis. Work on human placental ultrastructure was initiated at Cambridge and Kyoto whilst domestic animals were initially studied by Björkman in Stockholm and electron micrographs of bat placenta were published by Wimsatt of Cornell University. CONCLUSIONS: Prior to the introduction of better fixation techniques, TEM images were of modest technical quality. Nevertheless they gave important insights into placental ultrastructure, particularly the nature of the maternal-fetal interface.

Placentation in mammals: Definitive placenta, yolk sac, and paraplacenta.

An overview is given of variations in placentation with particular focus on yolk sac, paraplacenta, and other structures important to histotrophic nutrition. The placenta proper varies in general shape, internal structure, and the number of tissues in the interhemal barrier. Yolk sac membranes persist to term in insectivores, colugos, rodents, and lagomorphs. In the latter two orders, they are of known importance for maternal-fetal transfer of antibodies, vitamins, lipids, and proteins. The detached yolk sac of bats is also active throughout gestation. A vascular paraplacenta, or smooth chorioallantois, has known functions in ruminants and carnivores and is found in several other orders of mammal where its function has yet to be explored. In human gestation, the chorion (avascular chorioallantois) is important for hormone synthesis. The true chorion of squirrels and hedgehogs is avascular but may nevertheless allow transfer from mother to fetus through the exocelom. Hemophagous areas with columnar trophoblast are paraplacental structures in carnivores and elephants but occur also within the placenta as in hyenas and moles. In shrews, it is the yolk sac that ingests and processes red cells. Areolas and chorionic vesicles are other structures important for absorption of uterine secretions and ingestion of cellular debris. In conclusion, we find that paraplacental structures, while showing less variation than the placenta proper, contribute not just to the integrity of overall placentation, but in various ways to maternal-fetal interrelationships.

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.

Interstitial trophoblast cells: an enigmatic and variable component of the developing macaque placenta.

The distribution of cytokeratin-positive interstitial trophoblast cells in the endometrium of the macaque during placental development was examined. Such cells are moderately abundant only from the 15th through the 22th day of pregnancy, although there is considerable individual variation. During this period of gestation, interstitial trophoblast cells are distributed in the perivascular stroma immediately surrounding spiral arteries, including coils of arteries already invaded by endovascular trophoblast. The interstitial trophoblast cells are not seen to directly intrude into the smooth muscle of the spiral arteries. Very few interstitial trophoblast cells are present from days 12 through 14 of gestation when the arteries are first invaded by endovascular trophoblast. Even fewer interstitial trophoblast cells are seen after day 30 of gestation. The brief time in gestation when interstitial trophoblast cells are abundant coincides with a time when spiral arteries are undergoing rapid modification into patent uteroplacental arteries. It is suggested that during this period, the interstitial trophoblast cells may facilitate changes in shape of the spiral arteries by alterations of the perivascular connective tissue, perhaps through interactions with other cellular constituents rather than by directly modifying the arteries per se.

The evolving placenta: convergent evolution of variations in the endotheliochorial relationship.

Endotheliochorial placentas occur in orders from all four major clades of eutherian mammal. Species with this type of placenta include one of the smallest (pygmy shrew) and largest (African elephant) land mammals. The endotheliochorial placenta as a definitive form has an interhemal area consisting of maternal endothelium, interstitial lamina, trophoblast, individual or conjoint basal laminas, and fetal endothelium. We commonly think of such placentas as having hypertrophied maternal endothelium with abundant rough endoplasmic reticulum (rER), and as having hemophagous regions. Considering them as a whole, the trophoblast may be syncytial or cellular, fenestrated or nonfenestrated, and there may or may not be hemophagous regions. Variations also appear in the extent of hypertrophy of the maternal endothelium and in the abundance of rER in these cells. This combination of traits and a few other features produces many morphological variants. In addition to endotheliochorial as a definitive condition, a transitory endotheliochorial condition may appear in the course of forming a hemochorial placenta. In some emballonurid bats the early endotheliochorial placenta has two layers of trophoblast, but the definitive placenta lacks an outer syncytial trophoblast layer. In mollosid bats a well developed endotheliochorial placenta is present for a short time even after a definitive hemochorial placenta has developed in a different region. It is concluded that the endotheliochorial placenta is more widespread and diversified than originally thought, with the variant with cellular trophoblast in particular appearing in several species studied recently.

Review: The evolving placenta: different developmental paths to a hemochorial relationship.

The way in which maternal blood is associated with trophoblast prior to the formation of the different types of hemochorial placenta may be conveniently grouped into four main patterns: a transitory endotheliochorial condition; maternal blood released into a mass of trophoblast; maternal blood confined to lacunae; and fetal villi entering preexisting maternal blood sinuses. Although it might be considered logical that developing placentas would pass through an endotheliochorial stage to become hemochorial, this developmental pattern is seen only as a transient stage in several species of bats and sciuromorph rodents. More commonly a mass of trophoblast at the junction with the endometrium serves as a meshwork through which maternal blood passes, with subsequent organization of a labyrinth when the fetal vascular component is organized. The initial trophoblast meshwork may be cellular or syncytial, often leading to a similar relationship in the spongy zone and labyrinth. Old World monkeys, apes and humans have a lacunar stage prior to establishing a villous hemochorial condition. New World monkeys lack a true lacunar stage, retaining portions of maternal vessels for some time and initially forming a trabecular arrangement similar to though differently arrived at than that in the tarsier. In armadillos, preexisting maternal venous sinuses are converted into an intervillous blood space by intruding fetal villi. Variations from the major patterns of development also occur. The way in which the definitive placental form is achieved developmentally should be considered when using placental structure to extrapolate evolution of placentation.

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.

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.

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.

Reasons for diversity of placental structure.

Many early embryonic stages are nearly indistinguishable in different Eutheria. However, implantation stages and placental morphological types vary tremendously. A number of factors favor this conflicting diversity. 1. Whereas embryo development takes place in the isolation of the amniotic cavity, the extraembryonic membranes of the conceptus develop in close association with the uterus of a genetically different organism. 2. Early conditions for the developing conceptus are anaerobic whereas later in development efficient aerobic conditions are essential for continued growth of the fetus. 3. Developing extraembryonic membranes have the potential to form two partially sequential placentas. The yolk sac can participate in forming a choriovitelline placenta, including an interhemal region, and can be adapted to various non-respiratory functions as gestation proceeds. Development of the chorioallantoic placenta begins later than the choriovitelline placenta but can overlap with this before supplanting it. The original development of the extraembryonic membranes occurs when the conceptus is sufficiently small that neither its nutritional requirements nor its respiratory needs are the burden to the maternal organism that they are later in gestation. Despite these developmental factors promoting different methods of forming the definitive placenta, the placental type is consistent within most families indicating that the divergence in placental structure accompanied evolution of differences between groups.

Female reproductive tract and placentation in sucker-footed bats (Chiroptera: Myzopodidae) endemic to Madagascar.

The reproductive tract was examined in four non-pregnant and two gravid specimens of Myzopoda. The ovaries had little interstitial tissue. The uterus was bicornuate and the lenticular placental disk was situated mesometrially in one horn. The interhaemal barrier of the placental labyrinth was of the endotheliomonochorial type. There was a single layer of trophoblast cells. The cells of the maternal endothelium were large and basophilic, contained abundant rough endoplasmic reticulum, and rested on an irregular basement membrane. Blunt projections of endometrium extended into the placental disk and clusters of large cells occurred between the endometrial stroma and labyrinth. At the margins of the disk folds of trophoblast occurred and at the cranial end they formed an haemophagous region. The folds lateral to the disk included some peculiar tubular-appearing structures. There was a persistent yolk sac containing large endodermal cells around a largely collapsed lumen. Several features of placentation, such as the interhaemal barrier and the haemophagous region, are consistent with an association of Myzopodidae with Emballonuridae. No support was found for alternative hypotheses that include Myzopodidae in the noctilionoid or vespertilionoid lineages.

Placental diversity in malagasy tenrecs: placentation in shrew tenrecs (Microgale spp.), the mole-like rice tenrec (Oryzorictes hova) and the web-footed tenrec (Limnogale mergulus).

Placentation in tenrecs of the subfamily Oryzorictinae, family Tenrecidae, has not been described previously. The structure of the placenta of this group and especially of the genus Microgale was investigated to determine its similarity or dissimilarity to previously described placentas of the tenrec subfamilies Potamogalinae and Tenrecinae. Fifteen specimens of the genus Microgale ranging from an early yolk sac stage to near term were available for study. Placentation in Microgale was found to be different from other tenrecids in that there is an early simple lateral rather than central haemophagous region. In addition, a more villous portion of the placental disk forms before the formation of a more compact labyrinth. Although the definitive placenta is cellular haemomonochorial, it lacks the spongy zone found in the Tenrecinae. Neither does it resemble the endotheliochorial condition found in the Potamogalinae. Of the two genera of the subfamily Oryzorictinae represented by single specimens, the placenta of Limnogale resembled that of the Microgale but Oryzorictes had several differences including a lobulated placental disk. It is concluded that there is more variation in placentation both within the subfamily Oryzorictinae and within the family Tenrecidae than would ordinarily be expected.

Implantation in the macaque: expansion of the implantation site during the first week of implantation.

Data accumulated over several years of investigating implantation in macaque monkeys have been used to estimate the rate of expansion of the initial implantation site, the increase in volume of the site, and the rate of arterial invasion by cytotrophoblast columns. In addition the expansion of the secondary implantation site has also been estimated. The primary implantation site expands from an average diameter of 0.268 mm on day 10 to 4.93 mm on day 16-17. It expands in thickness from 0.064 mm on day 10 to 0.96 mm on day 16-17, and in volume from 0.0036 mm(3) on day 10 to 18.34 mm(3) on day 16-17. During this period of rapid expansion in extent and volume of the implantation site, trophoblast invades the endometrium, forms a lacunar stage, and initiates villus formation; consequently these very considerable changes in structure occur when the implantation is still very small yet growing rapidly. The secondary site expands from 0.23 mm in diameter on day 12 to 2.8mm on day 17. The rate of penetration of cytotrophoblast into endometrial arteries diminishes from 0.602 mm per day on day 12 to 0.171 mm per day on day 16, using the straight-line method of estimation. This diminution in rate is consistent with the hypothesis that cytotrophoblast cells generated at the anchoring villi migrate over cytotrophoblast cells that have attached to the endothelium of the endometrial arteries in advancing the intraluminal columns of trophoblast. It is hoped that the summaries provided will be useful to investigators using macaque monkeys to analyze aspects of implantation in primates.

Comparative placentation: some interesting modifications for histotrophic nutrition -- a review.

In considering the diversity of Eutherian mammalian placental structure, it is helpful to keep in mind that both phylogenetically and ontogenetically a functional yolk sac placenta precedes development of the chorioallantoic placenta. Usually the chorioallantoic placenta progressively displaces the area of contact of the yolk sac placenta with the endometrium. It is also closely applied to the endometrium, increasing respiratory efficiency but tending to decrease histotrophic nutrition. Catarrhine primates have minimal histotrophic uptake, using haemotrophic mechanisms throughout most of gestation. Rodents, by using partial or complete inversion of the yolk sac, have extensive regions available to histotroph in later pregnancy. Most mammals, however, have developed specialized regions of the chorioallantoic placenta for ingestion of uterine secretions, cell debris and erythrocytes. These regions vary from simple saccular areolae to more complex areolae such as the chorionic vesicles of prosimian primates to specialized haemophagous areas. All such structures are consistently lined by columnar cellular trophoblast. Examples of these structures, many of which can ingest both glandular secretions and erythrocytes (i.e. they are heterophagous), can be found in species whose definitive placenta is epitheliochorial. They are common but not universal in species whose definitive placenta is endotheliochorial and are even found in a few species with definitive haemochorial placentas. Restriction of phagocytosis to regions of columnar cells provides polarized cells appropriately oriented for ingestion, breakdown and transport, and limits the type of ingestion of both beneficial and potentially toxic materials to expendable individual cells.

Suppression of extravillous trophoblast invasion of uterine spiral arteries by estrogen during early baboon pregnancy.

The present study determined whether estrogen plays a role in regulating invasion and remodeling of the uterine spiral arteries by extravillous trophoblasts during early baboon pregnancy. The level of trophoblast invasion of spiral arteries was assessed on day 60 of gestation (term is 184 days) in baboons untreated or treated on days 25-59 with estradiol or aromatizable androstenedione. The administration of estradiol or androstenedione increased (P<0.01) maternal serum estradiol levels approximately 3-fold above normal. The mean+/-SE percentage of spiral arteries/arterioles invaded by extravillous cytotrophoblasts in estradiol-treated baboons for vessels with diameters of 26-50 microm (0.0+/-0.0), 51-100 microm (1.2+/-0.7) and >100 microm (13.2+/-5.5) was 100%, 90%, and 75% lower (P<0.001), respectively, than in untreated baboons (2.4+/-1.2%; 11.0+/-5.5%, and 54.5+/-8.5%, respectively). Similar results were obtained with androstenedione treatment. However, the distribution of uterine spiral arteries grouped by diameter or number of arteries per basal plate area, i.e. microvessel density, were similar in untreated and estrogen-treated baboons. We suggest, therefore, that the low levels of estrogen exhibited during early primate pregnancy are required to permit normal progression of trophoblast vascular invasion and that the surge in estrogen which occurs during the second-third of normal pregnancy has a physiological role in suppressing further arterial trophoblast invasion. Consequently, we propose that the estrogen-dependent restraint of spiral artery invasion/remodeling ensures optimal blood flow dynamics across the uteroplacental vascular bed to promote normal fetal growth and development.

The fetal membranes of the otter shrews and a synapomorphy for afrotheria.

The otter shrews of mainland Africa are the closest relatives of the Madagascar tenrecs. We sought for similarities in placentation between the two groups and, in a wider context, with other mammals of the Afrotheria clade. Specimens of the Nimba otter shrew (Micropotamogale lamottei) were obtained from the Ivory Coast and examples of the giant otter shrew (Potamogale velox) from the Hill Collection. The Nimba otter shrew has a central haemophagous organ similar to that in tenrecs. The labyrinth of the Nimba otter shrew, however, is endotheliochorial with syncytial trophoblast enclosing the maternal vessels. On the other hand tenrecs have cellular haemomonochorial placentae and an associated spongy zone, which is not present in the Nimba otter shrew. The placenta of the giant otter shrew is also endotheliochorial. The central region of its placenta is particularly interesting, since the juxtafetal portion is clearly a haemophagous region whereas the labyrinth feeding this region is endotheliochorial. Thus there is considerable variation in placental morphology within Tenrecidae. Importantly, however, both otter shrews have a large allantoic sac divided into four intercommunicating lobes by two pairs of septal folds. A similar arrangement has been described for representatives of each of the remaining five orders within Afrotheria. This is significant because previous anatomical studies have failed to establish a single synapomorphy in support of Afrotheria.

Structure of the ovaries of the Nimba otter shrew, Micropotamogale lamottei, and the Madagascar hedgehog tenrec, Echinops telfairi.

The otter shrews are members of the subfamily Potamogalinae within the family Tenrecidae. No description of the ovaries of any member of this subfamily has been published previously. The lesser hedgehog tenrec, Echinops telfairi, is a member of the subfamily Tenrecinae of the same family and, although its ovaries have not been described, other members of this subfamily have been shown to have ovaries with non-antral follicles. Examination of these two species illustrated that non-antral follicles were characteristic of the ovaries of both species, as was clefting and lobulation of the ovaries. Juvenile otter shrews range from those with only small follicles in the cortex to those with 300- to 400-microm follicles similar to those seen in non-pregnant and pregnant adults. As in other species, most of the growth of the oocyte occurred when follicles had one to two layers of granulosa cells. When larger follicles became atretic in the Nimba otter shrew, hypertrophy of the theca interna produced nodules of glandular interstitial tissue. In the tenrec, the hypertrophying theca interna cells in most large follicles appeared to undergo degeneration. Both species had some follicular fluid in the intercellular spaces between the more peripheral granulosa cells. It is suggested that this fluid could aid in separation of the cumulus from the remaining granulosa at ovulation. The protruding follicles in lobules and absence of a tunica albuginea might also facilitate ovulation of non-antral follicles. Ovaries with a thin-absent tunica albuginea and follicles with small-absent antra are widespread within both the Eulipotyphla and in the Afrosoricida, suggesting that such features may represent a primitive condition in ovarian development. Lobulated and deeply crypted ovaries are found in both groups but are not as common in the Eulipotyphla making inclusion of this feature as primitive more speculative.

Development of the haemophagous region and labyrinth of the placenta of the tenrec, Echinops telfairi.

Our purpose was to determine how the central haemophagous region and cellular haemomonochorial labyrinth of the tenrec placenta are formed. The haemophagous region is preceded by a region of invasion of the endometrium by trophoblast comprising a cytotrophoblast layer covered by syncytial trophoblast and contiguous with numerous masses of multinucleate trophoblast. The trophoblast intrudes into the endometrium, eliminating the stroma, although small vessels and clumps of glandular epithelium persist. This extensive central region is connected to the forming disk by a ring of chorioallantois covered by a single layer of columnar trophoblast. Later the multinucleate masses and syncytial trophoblast degenerate. The unilaminar cytotrophoblast remains, is elaborated into folds, and phagocytoses glandular secretion, cell debris and erythrocytes. As the central area is transforming, fetal capillaries move into the cytotrophoblast pads surrounding the central zone. Prior to this, the cytotrophoblast has formed a multilayered structure and interrupted maternal vessels to create an anastomotic network of blood spaces lined by cytotrophoblast. The invasion of fetal capillaries transforms this preplacental pad into a cellular haemomonochorial labyrinth with the uninvaded portion forming an underlying spongy zone. Thus interaction of the trophoblast with the endometrium is substantially different in the central zone compared to the area of the preplacental pad.