Reproductive system development in male and female horse embryos and fetuses: Gonadal hyperplasia revisited.

In horses, pregnancy is characterized by high levels of maternal estrogens that are produced largely by the interstitial tissue inside the gonads of the offspring, associated with a physiological gonadal hyperplasia, that is uncommon in other species. However, a detailed structural-functional understanding of the early stages of gonadal development and hyperplasia has remained elusive in horse pregnancy because of the lack of substantial data. The goal of this study was to describe the genital organs' development in 19 early horse embryos and fetuses (days 20-140 of gestation) of both sexes by means of anatomy, histology, stereology, and immunohistochemistry, with a specific focus on gonadal hyperplasia and interstitial tissue development. Gonadal hyperplasia with similar amounts of interstitial cells was observed in both sexes, but only during the early stage of development (days 40-90). Surprisingly, a higher degree of hyperplasia, characterized by larger amounts of interstitial cell-rich areas, was seen in fetal ovaries from 90 days of gestation onwards. Another novel aspect was that parallel to the hyperplasia of the interstitial cells, a much more precocious and pronounced differentiation of germinal cells was seen in the ovary, characterized by an earlier peak and decrease of DAZL and OCT protein immune markers. In conclusion, a reduced degree of hyperplasia and interstitial tissue in the fetal testis after 90 days of gestation suggests the existence of a more efficient mechanism regarding the synthesis of estrogen precursors as a structural or physiological difference between both fetal sexes, which warrants further investigation.

Placentation in the colugos Cynocephalus volans and Galeopterus variegatus (Dermoptera) and the transition from labyrinthine to villous placentation in primates.

INTRODUCTION: Phylogenetics and genomics place colugos as the sister group to primates. Therefore their placentation is of interest in an evolutionary perspective. Previous accounts are fragmentary, not readily accessible and sometimes contradictory. METHODS: We have examined archival material covering the early development of fetal membranes and placenta, the fate of the yolk sac and definitive placentation. RESULTS: Initially the trophoblast extended over a rather broad but shallow area, enclosing maternal blood spaces. After expansion of the exocoelom it became covered by somatic mesoderm. The mature chorioallantoic placenta was haemochorial and characterized by a labyrinth with markedly dilated maternal blood spaces. Blood vessels appeared in the splanchnopleure early in development and later extended to the yolk sac, but we found no evidence of a choriovitelline placenta at any stage of gestation. There was, however, an extensive paraplacenta. CONCLUSIONS: A choriovitelline placenta is not formed early in gestation nor is it present at term. Early in development invasive trophoblast spreads laterally to form a trophoblastic plate. We found evidence to support the idea that the colugo placenta is intermediate between the labyrinthine placenta of rodents and the trabecular type of Neotropical primates.

Extracellular matrix in epitheliochorial, endotheliochorial and haemochorial placentation and its potential application for regenerative medicine.

A placenta is defined as structural approximation of maternal and foetal tissues to perform physiological exchange. Associated processes of differentiation and the establishment of its cells take place within the extracellular matrix (ECM) that provides a rich environment of collagens, fibronectins, cytokines and other components. Placental ECM is promising for tissue regeneration purposes, because it has immune tolerance capacities that may cause only minimal rejections of transplants with immunological differences between donor and recipient. However, specific characteristics of ECM during evolution of the structurally very diverse mammalian placenta are not yet revealed. We here address the major aspects of placental types, that is non-invasive (epitheliochorial), medium (endotheliochorial)-to-high (haemochorial) invasive nature of the interhemal barrier between the foetal and maternal blood system as well as their main components of ECM with special reference to species that are commonly used as animal models for human placentation and in the potential applications for regenerative medicine.

Development of yolk sac inversion in Galea spixii and Cavia porcellus (Rodentia, Caviidae).

Caviomorph development includes an inverted yolk sac. Since principle processes are not understood, we investigated its differentiation in Galea and re-examined material from the guinea pig. Galea showed the typical caviomorph conditions in blastocyst development and the nature of the definitive yolk sac, formed of the visceral layer that became villous, proliferative, vascularized and attached to the uterus and placenta. In contrast to what was known before, in both species parts of the parietal yolk sac and a yolk sac cavity were temporarily present. Data suggest that early yolk sac development in caviomorphs is more complex than thought before.

An unusual feature of yolk sac placentation in Necromys lasiurus (Rodentia, Cricetidae, Sigmodontinae).

We studied the development of the inverted yolk sac in a New World rodent, Necromys lasiurus during early placentation. Ten implantation sites were investigated by means of histology, immunohistochemistry and electron microscopy. The yolk sac was villous near its attachment to the placenta. Elsewhere it was non-villous and closely attached to the uterus. The uterine glands were shallow and wide mouthed. They were associated with vessels and filled with secretion, suggesting the release of histotroph. This feature was absent at later stages. The intimate association of the yolk sac with specialized glandular regions of the uterus may represent a derived character condition of Necromys and/or sigmodont rodents.

Chorioallantoic and yolk sac placentation in the plains viscacha (Lagostomus maximus) - a caviomorph rodent with natural polyovulation.

OBJECTIVES: Reproduction in the plains viscacha is characterized by the polyovulation of hundreds of oocytes, the loss of implantation and the development of 1-3 offspring. Our goal was to determine whether placental development was affected by these specializations. STUDY DESIGN: Thirteen placentas from early pregnancy to near-term pregnancy were analyzed using histological, immunohistochemical and transmission electron microscopy. RESULTS: An inverted, villous yolk sac was present. Placentas were formed by the trophospongium, labyrinth and subplacenta. A lobulated structure with a hemomonochorial barrier was established early in pregnancy. Proliferating trophoblast that was clustered at the outer border and inside the labyrinth was responsible for placental growth. Trophoblast invasion resulted from the cellular trophoblast and syncytial streamers derived from the subplacenta. Different from other caviomorphs, numerous giant cells were observed. CONCLUSIONS: The principle processes of placentation in caviomorphs follow an extraordinarily stable pattern that is independent of specializations, such as polyovulation.

Hans Strahl's pioneering studies in comparative placentation.

Hans Strahl, a contemporary of Duval and Hubrecht, made many important contributions to comparative placentation. Despite this he is not well known and some of his original observations tend to be attributed to later authors. Strahl published a classification of placental types based on their shape and relationship to maternal tissues. This greatly influenced the work of Otto Grosser, who became better known in part because his work was more accessible to other scientists and clinicians. Strahl described the development of the fetal membranes across a broad range of mammalian orders extending his observations beyond parturition to the post partum involution of the uterus. He paid close attention to structures designed for histotrophic nutrition including the areolae of moles, haemophagous organs of carnivores and tenrecs and chorionic vesicles of lemurs and lorises. We here provide a summary of some of the most important findings made by Strahl including work on placentation in carnivores and higher primates that remains unsurpassed.

Evolution of the interhaemal barrier in the placenta of rodents.

A recent phylogenetic analysis achieved good resolution between the 5 suborders of rodent. As a novel finding it suggested a basal split that gave rise to a monophyletic group comprising Hystricomorpha and Sciuromorpha. We asked whether the new tree could cast light on the evolution of the interhaemal barrier in rodents where at least seven variants have been described. To supplement existing data we first examined the placenta of the common gundi, Ctenodactylus gundi. It was shown to be haemochorial with a single layer of syncytiotrophoblast in the interhaemal membrane but with nests of cytotrophoblast elsewhere. Next we used character mapping on the recent tree to determine the pattern of evolution of the placenta with respect to principal type (e.g. haemochorial) and the trophoblast found within the interhaemal barrier. This indicated that the common ancestor of living rodents had a haemochorial placenta and that there were two independent transformations to the endotheliochorial type. Moreover, the interhaemal barrier was found to have had a single layer of syncytial trophoblast in the common ancestor of rodents, a condition that was retained in the clade comprising Hystricomorpha and Sciuromorpha. In contrast the second clade shows multiple character transformations.

Development of the inverted visceral yolk sac in three species of caviids (Rodentia, Caviomorpha, Caviidae).

Guinea pig related rodents possess numerous derived placental characters. We attempt to identify diversity within the visceral yolk sac and its association with the chorioallantoic placenta in three species of caviids, two of them possessing a capsule formed by the decidua that covers the chorioallantoic placenta. The results verify that in early pregnancy all three species have an inverted yolk sac placenta. In advanced pregnancy the species differ: Galea spixii, as representative without a capsule, bear a yolk sac in apposition to the chorioallantoic placenta with signs of exchange activity until term. Galea is similar to other caviomorphs in this respect. In Dasyprocta leporina and Cuniculus paca, the representatives possessing a capsule, the yolk sac endoderm lacks signs of substance exchange. Evidently, the presence of a capsule prevents such an interaction. The variations established here must be considered if animal models for human placentation are required which have restricted access to the chorioallantoic placenta from the outside.

Caviomorph placentation as a model for trophoblast invasion.

The guinea pig and its relatives are promising candidates as animal models for studying trophoblast invasion. The origin, migration routes and kinetics of invasive trophoblast cells were examined in two caviomorph species. Histology and immunohistochemistry were done on placentas from 38 guinea pigs of days 20-47 and 13 degus of days 25-51 of gestation. BrdU was used as an in vivo marker for proliferation and for tracing of migration routes in the placenta; it was injected 24h to 15 days before collecting the material. In both species extravillous-like trophoblast cells are derived from proliferating stem cell aggregations in the subplacenta, which are comparable to the cell columns in humans. Migration routes and kinetics under in vivo conditions revealed a mean invasive depth of 300-350 microm/day and a mean life span of the extravillous-like trophoblast of 30 days. The patterns of trophoblast invasion in caviomorphs are analogous to the situation in humans, suggesting that these rodents are appropriate animal models for the study of the dynamics of trophoblast invasion.

Chorioallantoic and yolk sac placentation in the dassie rat Petromus typicus and its significance for the evolution of hystricognath rodents.

Placental characters are most important in understanding the evolutionary history of hystricognath rodents of which some act as animal models for human pregnancy. The data available deal mostly with species native to South America, but the current paper presents novel findings on chorioallantoic and yolk sac placentation in an Old World hystricognath and discusses its significance for the evolution of the group. Several hystricognath stem species characters are verified for Petromus, such as the unique trophoblast growth pattern within the chorioallantoic placenta. Subsequently, a novel set of characters belonging to the visceral yolk sac is added to the stem species pattern of the group. The nourishment of the embryo is facilitated by an inverted visceral yolk sac placenta from early pregnancy onward, later complemented by the chorioallantoic placenta. About mid term, the visceral yolk sac becomes partly folded and attached to the parietal yolk sac cover of the chorioallantoic placenta, suggesting a functional shift to the transfer of substances between the two placental types. Thus, the chorioallantoic and yolk sac placenta collaborate in nurturing the embryo. This apparently represents an evolutionary transformation along the stem lineage of hystricognaths.

The Guinea pig placenta: model of placental growth dynamics.

Placental growth dynamics are an important, but poorly understood aspect of placentation for which suitable animal models are lacking. Although the guinea pig is recognised as a valuable model for human pregnancy, its suitability for this purpose is not apparent, since the developmental and evolutionary processes that led to its lobulated placenta are not fully understood. Therefore, a study on immunohistochemistry has been undertaken, including the antibody mib-1 against protein Ki-67 as a proliferation marker. It was found that cellular trophoblast derived from the placental margin is the major centre of proliferation, supported by a moderate number of proliferating cells inside the labyrinth. In contrast to the ancestral condition of caviomorph rodents, the evolution of significant placental lobulation requires the latter source of proliferation. The distribution pattern of proliferating trophoblast cells revealed for the guinea pig strongly resembles the distribution in the human placenta in that (a) evenly distributed, moderate numbers of cells occur within the exchange area and (b) focal accumulations of proliferating trophoblast cells are present along the fetal mesenchymal border and appear to serve as a growth zone for appositional growth of new exchange tissues. Thus, the guinea pig appears to be an appropriate animal model for placental growth processes in human pregnancies.

Evolutionary differentiation of Cetartiodactyl placentae in the light of the viviparity-driven conflict hypothesis.

We analysed the evolution of placental traits in the novel mammalian clade Cetartiodactyla (Cetaceans and Artiodactyls) by a parsimony-based computer program (MacClade). A diffuse epitheliochorial placenta was identified as the stem species pattern of this clade. Trophoblast giant cells (TGCs) independently evolved in Camelids and Ruminants. The polycotyledonary placenta is an apomorphic character for Pecora (higher ruminants) and the oligocotyledonary placenta developed as a further step on the stem lineage of cervidae and moschidae. We interpret these findings by application of the "viviparity-driven conflict hypothesis", which states that divergent interests of mother and offspring lead to a rapid antagonistic coevolution, which might cause placental diversity. According to this hypothesis the evolution of camelid and ruminant TGCs can be interpreted as means to increase fetal endocrine influence on the maternal metabolism. The development of the cotyledonary placenta could be related to a diminished availability of glucose, which is associated with the evolution of forestomach fermentation in Pecora. An arms race, in which the mother tried to restrict and the fetus tried to increase transplacental glucose flow, might have promoted the evolution of the cotyledonary placenta, which has a high feto-maternal exchange area, but a low conductivity for glucose.

Evolution of the placenta in eutherian mammals.

We review recent advances in our understanding of placental evolution with particular focus on the interhaemal barrier. It seems likely that the non-invasive, epitheliochorial placentation of living eutherians is a derived state. However, there is disagreement on whether the last common ancestor of eutherian mammals had an endotheliochorial placenta or a haemochorial one. Research has been stimulated by improved understanding of the relations between the orders of mammals provided by molecular phylogenetics. In part, the uncertainties arise from doubt about how to root the mammalian tree. Resolution of this issue will require improved taxon sampling in molecular analyses. At the same time, we need to foster research in comparative placentation on relevant taxa, particularly at the ultrastructural level. Only then can we ensure that information available about the placenta is adequate to capitalise on future advances in molecular phylogenetics. Examples are given of recent findings that could benefit cladistic analysis of placental evolution.

Mesotocin gene expression in the diencephalon of domestic fowl: cloning and sequencing of the MT cDNA and distribution of MT gene expressing neurons in the chicken hypothalamus.

This study focuses on the structure and expression of the mesotocin (MT) gene in the chicken hypothalamus. Using an anchored and nested RT-PCR strategy, combined with circular RACE-PCR, the full length sequence of the chicken MT cDNA was obtained. The cDNA and derived amino acid sequences conformed to the structure of the oxytocin-like gene family. However, unlike most mammalian species, there does not appear to be frequent gene conversion between the MT and AVT cDNA sequences. A single specific hybridization signal of 1.2 kb was detected by Southern analysis of chicken genomic DNA, indicating only a single gene copy in the chicken genome. Northern analysis of hypothalamic RNA revealed a single band at approximately 0.6 kb. Using the same probe for in situ hybridization histochemistry, MT-mRNA was demonstrated to be predominantly localized in the parvocellular, magnocellular and periventricular subgroups of the paraventricular nucleus and, when compared to the distribution of neurons containing arginine-vasotocin (AVT)-mRNA in the same region, with far fewer neurons expressing the MT gene in the lateral subgroups. Only few and scattered neurons expressing the MT gene were found in the ventral and external subgroups of the supraoptic nucleus in which many neurons contain AVT transcripts, as demonstrated in consecutive sections. In all nuclei investigated, the intensity of AVT and MT hybridization signals per cell was approximately equal. No specific labelling for MT-mRNA was found in the bed nucleus of the stria terminalis, nor the nucleus accumbens. Using immunocytochemical detection of AVT and in situ hybridization for neurons expressing MT-mRNA, some neurons were found to contain both AVT and MT gene products in the paraventricular nucleus but not in the supraoptic nucleus.