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.     

不同产羔数西农萨能奶山羊胎盘性状的差异分析

为研究不同产羔数西农萨能山羊胎盘性状的差异,本实验共收集73只(单羔24只;双羔42只;三羔7只)正常分娩的西农萨能奶山羊母羊胎盘,分析比较不同产羔数的胎盘效率(初生窝重与胎盘质量之比)、子叶承载效率(初生窝重与子叶总面积之比)、子叶密度(子叶总数与胎盘质量之比)、子叶面积等性状和子叶组织学结构与母羊繁殖性能之间的关系。结果表明:随产羔数增加,西农萨能山羊胎盘质量呈极显著增加;多羔组胎盘子叶总数、子叶总面积和子叶承载效率显著高于单羔组;组织学结构分析发现,胎盘子叶滋养层细胞数量、毛细血管数量及密度也随着产羔数增加而增加。因此,胎盘质量、子叶总数和子叶总面积等性状可能是高繁殖力母羊的选择指标之一。     

Morphometric study of the porcine placental vascularization

The early development in mammals is characterized by the contribution of nutrients from the maternal tissues through the placenta, which is in apposition with foetal membranes and the endometrium, allowing the physiological interchange between the embryos/foetuses and the mother. The aim of this work was to study the number of placental blood vessels and their vascular area through morphometric analyses and the haemotrophic diffusion distance in porcine placental tissues from early gestations, intermediates gestations, advanced gestations and term gestations. For those purposes, morphometric measurements, blood vessel quantification, high‐resolution light microscopy and transmission electron microscopy were performed. The implementation of the high‐resolution light microscopy allowed studying the placental vascular and tissue histoarchitecture with higher definition and resolution than using a conventional light microscopy. We highlight the close location of the subepithelial capillaries to the maternal/foetal interface as pregnancy progresses. We found statistically significant evidence to state that the area of blood vessels is dependent on the gestation period. In advanced gestations, the presence of numerous small blood vessels and its near location to foetal/maternal interface agree with the great remodelling reported in our previous studies. In conclusion, in gilts, given the type of non‐invasive epithelial placentation, the new blood vessels generation and of haemotrophic diffusion distance reduction, determined in this report, assure the maternal/foetal haemotrophic exchange efficiency during gestation.     

Effects of testosterone on rat placental development

We investigated the morphological effects of testosterone on placental development in a rat model of polycystic ovarian syndrome (PCOS). Testosterone propionate (TP), which was subcutaneously administered to pregnant rats with 5 mg/animal from gestation day (GD) 14 to GD 18, induced a maternal weight reduction without mortality or clinical signs from GD 19 onwards. A decrease in fetal and placental weight, an increase in intrauterine growth retardation (IUGR) rates, and histological changes in the placenta were observed on GD 21 but not on GD15 or 17. Histopathologically, on GD 21, the trophoblast septa thickened, and the maternal sinusoids were narrowed in the labyrinth zone, resulting in a small placenta. Additionally, the placental weight, thickness, and histological morphology in the labyrinth zone on GD 21 in the TP-treated group were nearly identical to those on GD 17 in the control and TP-treated groups. Therefore, it was assumed that the testosterone-induced small placenta was induced in association with the developmental inhibition of the fetal part of the placentas from GD 17 onwards.     

Gastrointestinal Diseases of Marsupials

This article focuses on gastrointestinal diseases that affect 3 of the most common marsupials treated by veterinarians: the red kangaroo (Macropus rufus), Bennett’s wallaby (Macropus rufogriseus), and the sugar glider (Petaurus breviceps). Other marsupial species maintained in captivity include the eastern gray kangaroo (Macropus gigantus), the tammar wallaby (Macropus eugenii), and the wallaroo (Macropus robustus). Most all of the disease conditions discussed in this article are also diagnosed in the lesser species listed earlier. Marsupials derive their name from the presence of the marsupial bones (ossa marsupialia), which serve as the surface for the attachment of several abdominal muscles. The ossa marsupialia were once thought to support the marsupium (pouch), although this was an incorrect assumption. The metabolic rate of marsupials is generally considered to be approximately two-thirds that of eutherian or placental mammals. Marsupials belong to a unique group of animals whose development is characterized by a very short gestation, the birth of relatively underdeveloped young (joeys), and a lengthy period of lactational development that typically occurs in a pouch. The physiologic differences that exist between marsupials and eutherian mammals should be considered when managing disease conditions in these animals.     

Light and scanning electron microscopic study on the blood vascular system of the donkey placenta

The donkey placenta is diffuse and epitheliochorial with numerous microplacentomes consisting of a fetal microcotyledonary and a maternal microcaruncular part. The microplacentomal vasculature during the last third of pregnancy has been investigated by light microscopy in comparison to scanning electron microscopy of the materno-fetal contact surface and corrosion casts of blood vessels after plastic instillation from either the microcotyledonary or the microcaruncular side, and, for the first time in a perissodactyle, from both sides. Morphological data were semiquantitatively evaluated. The supplying parts of both, the microcotyledonary and the microcaruncular vascular system are strictly proximo-distally oriented, thus reaching the capillary systems or working parts in the shortest way possible. The straight course of the vasculature, particularly on the fetal side, suggests the occurrence of venulo-arteriolar back diffusion. The fetal capillary system consists of convolutes confronting the maternal septal capillary complexes in a countercurrent way. This materno-fetal blood flow interrelationship is highly efficient in terms of placental exchange, which is further supported (1) by dilations and increasing coiling of the fetal venular capillary limbs in particular and (2) by a decrease in the interhaemal distance from 12.5 to 7.2 microm between the two capillary systems. Besides the countercurrent blood flow interrelationship, some maternal branch arterioles reach the septal capillary system from the maternally oriented pole of the microplacentome or microcaruncle, respectively, resulting in the less efficient crosscurrent blood flow. Hence, in the donkey placenta fetal and maternal blood vessels meet in a mix of countercurrent and crosscurrent flow patterns.     

Abnormal Expression of TIMP-2, SOD, Vimentin and PAI Proteins in Cloned Bovine Placentae

Cloned mammals suffer from high rates of placental abnormality and foetal loss during pregnancy. We previously used 2-D gel electrophoresis and mass spectrometry for global proteomic analysis of cloned and normal bovine placentae to identify differential protein expression patterns. Here, we used Western blot analysis to confirm the expression levels of several pregnancy-related proteins putatively identified as being differentially expressed in somatic cell nuclear transfer (SCNT) vs normal bovine placentae. The expression levels of tissue inhibitor of metalloproteinase-2 (TIMP-2), its downstream protein, matrix metalloproteinase-2 (MMP-2), superoxide dismutase (SOD), vimentin and plasminogen activator inhibitor-1 (PAI) were analysed in the placentae of SCNT cloned Korean native cattle that died immediately after birth and in normal placentae obtained by AI. Our results revealed that TIMP-2 and SOD were up-regulated in SCNT placenta compared with normal placenta, whereas MMP-2 levels were comparable in cloned and normal placentae, and vimentin and PAI were significantly down-regulated in SCNT compared with normal placentae. Our results suggest that key proteins of placental development are abnormally expressed in SCNT cloned bovine placentae, probably resulting in abnormal placental function and clonal mortality.     

Light and scanning electron microscopy of the olfactory mucosa in the rufous sengi (Elephantulus rufescens)

Sengis are eutherian insectivores belonging to superorder Afrotheria, a recently defined clade of mammals that diverged from other placentals over 100 million years ago. In this study, a histological and ultrastructural analysis was carried out on the olfactory mucosa (OM) of the rufous sengi (Elephantulus rufescens) and the data were compared with those reported earlier in the dog (Canis familiaris) and the sheep (Ovis aries), whose dietary lifestyles are carnivorous and herbivorous, respectively. Qualitatively, the microstructure of the sengi's OM was basically similar to that of the other eutherian mammals except for differences in the pattern of cilia projection from the dendritic knobs of the olfactory receptor neurons (ORNs) and distribution of Bowman's glands within the lamina propria of the OM. On morphometry, significant differences (p < .05) were recorded with respect to olfactory epithelial (OE) thickness between the sengi (65.4 ± 2.6 μm) and the other species. ORN packing density and cilia number/ORN knob varied markedly only between the sengi (73.8 ± 5.4 mm^?2 × 10³ and 15 ± 4, respectively) and the sheep. No remarkable differences were noted in regard to ORN bundle diameters between sengis (62.7 ± 12.5 μm) and the other species. The observed differences in OM structural refinement may be attributed to olfactory function demand levels related to feeding lifestyles and ecology. Myrmecophagous insectivory, social monogamy, absentee maternal care and exposed sheltering habits are behavioural features that may warrant substantial OM modification in sengis.     

Fatty acid metabolism by the porcine placenta

Two experiments were performed to determine whether esterification is a major pathway of fatty acid utilization within porcine placenta and to determine what metabolic parameters may limit fatty acid transfer to the fetal pig. Maternal (endometrium) and fetal (chorioallantois) placenta were obtained by Caesarean section at d 110 of gestation in both experiments. Eight gilts were used in the first experiment. Tissue sections were incubated with palmitate at concentrations ranging from .25 to 2.0 mM. Maternal placenta metabolized palmitate at a higher rate than fetal placenta, although fetal placenta was more efficient in esterifying palmitate. Esterification composed the majority of palmitate utilization within fetal and maternal placenta. The second experiment evaluated the effect of dietary lipid on placental fatty acid metabolism and evaluated the ability of placenta to mobilize lipids. Fourteen gilts were divided into two groups of seven and fed a diet containing 15% tallow diet or a diet not supplemented with tallow (control) from d 90 to 110 of gestation. Dietary lipid had no detectable effects on lipoprotein lipase activity, [14C]palmitate metabolism, or lipolysis by the maternal or fetal placenta. Lipolytic activity of placental tissues was minimally affected by incubation with various proposed lipolytic activity of placental tissues was minimally affected by incubation with various proposed lipolytic agents. The data indicate that supply of fatty acids to the fetal pig may be limited by transfer of plasma fatty acids into the cytoplasm of placental cells or by regulatory enzymes for intermediate esterification; both types of limitations have been proposed to be influenced by fatty-acid binding proteins.     

Immunohistochemical detection of the orexin system in the placenta of cats

The aim of the present study was to investigate the presence and distribution of cells containing orexin A (OXA), and orexin type 1 and 2 receptors (OX1R and OX2R, respectively) in the feline placenta by means of immunohistochemical technique. OXA was identified in several decidual and syncytiotrophoblastic cells present in the lamellar portion of the placenta. In the same placental structures, few decidual and syncytiotrophoblastic cells showed the presence of OX1R-like immunoreactivity. Characteristically, immunopositivity for OX2R, but not for OX1R, was evidenced in the cells of the glandular layer. The orexinic system was not expressed in the uterine structures that were not engaged by the chorion. Our results provide the first evidence of the presence of a placental orexinic system in a mammalian species. Orexin A and both OX1R and OX2R are unequally distributed within the cat placenta. Local OXA production and the presence of specific receptors, differentially expressed in the placental structures of the cat, suggest that the orexinic system may participate in placental growth and development as well as in the regulation of its steroidogenic capacity via endocrine, paracrine and/or autocrine mechanisms.     

Gross and histological findings in the canine placenta and amnion at term: What’s normal,abnormal or pathological?

The canine placenta is an underexamined organ. Placental abnormalities can affect foetus development and may be responsible for a low weight of the infant at birth; however, knowledge on their clinical significance in the canine species is limited. We aimed to describe macroscopic and microscopic findings in the canine placenta and amnion at term in clinically uncomplicated pregnancies and to evaluate their relationship with birth weight of healthy puppies. During natural delivery or C-section, the birth weight of 82 puppies was recorded, 72 placentas and 66 amnions were recovered. The foetal and maternal surfaces of the placental girdle, marginal haematoma and amnion were evaluated. Each gross finding was recorded, morphometrically assessed and sampled for histological diagnosis. Furthermore, specimens of placenta and amnion were collected from representative areas and microscopic deviations from normal structure were evaluated in haematoxylin and eosin sections. Gross examination revealed ‘abnormalities’ in the 75.4% of the collected placentas. Necrosis was the gross change most commonly observed in the placental girdle (72.5%). Congestion (17.4%) and clotted blood/fibrinoid material (2.9%) were also observed. No gross changes of either the marginal haematoma or the amnion were recorded. Histologically, placental girdle showed necrosis (62.3%), mineralization (52.2%), congestion (36.2%) and neutrophilic infiltration (27.5%). Marginal haematoma exhibited mineralization (11.6%) and neutrophils (29%), while necrotic foci were rarely observed (4.3%). In the amnion, the most frequent alteration observed was hypertrophy of the epithelium (35.9%) followed by oedema (31.2%), mineralized foci (28.1%), fibrosis (23.4%), congestion (15.6%) and more rarely neutrophils (12.5%). Puppies’ birth weight was not statistically affected by either gross or histological abnormalities. Our study revealed that macroscopic and microscopic ‘abnormalities’ of the placenta and amnion may be common in uncomplicated pregnancies at term; however, no implications on puppies’ birth weight were observed. Deviations from ‘normal’ morphology of canine foetal adnexa warrant further investigation to assess their clinical implications if present.     

Structural determination of the oligosaccharides in the milk of a giant anteater (Myrmecophaga tridatyla)

There appears to be strong evidence supporting four major clades of placental mammals: (i) Afrotheria; (ii) Xenarthea; (iii) Euarchontoglires; and (iv) Laurasiathera. This is the first study in which the carbohydrates have been analyzed in the milks of Xenarthea. Our aim was to clarify whether the saccharides of giant anteater milk resemble those of other eutherian species, especially of the Asian elephant, which is one of the Afrotherea. We found that lactose is a dominant saccharide in this milk, as it is in that of most eutherians, and that the milk contains smaller amounts of oligosaccharides, whose structures were determined as Gal(α1‐3)Gal(β1‐4)Glc (isoglobotriose), Neu5Ac(α2‐6)Gal(β1‐4)Glc (6'‐N‐acetylneuraminyllactose), Neu5Gc(α2‐6)Gal(β1‐4)Glc (6'‐N‐glycolylneuraminyllactose), Neu5Ac(α2‐6)Gal(β1‐4)GlcNAc(β1‐3)Gal(β1‐4)Glc (LST c) and Neu5Gc(α2‐6)Gal(β1‐4)GlcNAc(β1‐3)Gal(β1‐4)Glc. This pattern of milk carbohydrates is rather close to that of the milks of other eutherian species. It is notable that the milk contained oligosaccharides with Neu5Gc residues but no oligosaccharides containing α(2‐3) linked Neu5Ac residues such as Neu5Ac(α2‐3)Gal(β1‐4)Glc.     

Experimental placental transfer of foot-and-mouth disease virus in mice.

An attenuated type O foot-and-mouth disease (FMD) virus which was virulent for infant, but not for pregnant, mice proved to be superior to a virulent type C FMD virus in the development of a model system for the study of placental transfer of FMD in mice. When mice were inoculated at day 8 or 12 of gestation with type O FMD virus, the virus was detectable in the maternal pancreas for 3 days and in the placenta for 6 days. Viral levels in the fetus and the amniotic fluid were inconsistent and were apparently due to a spillover from the placental infection. The elimination of the virus from the placenta coincided with the expected production of maternal 7S antibody. Mice inoculated from days 0 to 12 of gestation did not have a significant increase in dead young by day 18 (the day of necropsy). Similarly inoculated mice, when permitted to go to term, produced and raised normal-size litters. Inoculation on day 15 of gestation resulted in an increased number of deaths due to morbidity of the dams. It was concluded that the placenta serves as an active site of infection for FMD virus in pregnant mice, but the fetus is relatively resistant to infection.     

Transplacental Transfer of Iron in the Water Buffalo (Bubalus bubalis): Uteroferrin and Erythrophagocytosis

The objectives of this investigation were to understand transplacental transport of iron by secreted uteroferrin (UF) and haemophagous areas of water buffalo placenta and clarify the role(s) of blood extravasation at the placental‐maternal interface. Placentomes and interplacentomal region of 51 placentae at various stages of gestation were fixed, processed for light and transmission electron microscopy, histochemistry and immunohistochemistry. Haemophagous areas were present in placentomes collected between 4 and 10 months of pregnancy. Perl’s reaction for ferric iron was negative in placentomes, but positive in endometrial glands. Positive staining for UF indicated areas in which it was being taken up by phagocytosis and/or fluid phase pinocytosis in areolae of the interplacentomal mesenchyme, with little staining in endometrial stroma. Imunohistochemistry detected UF in trophectoderm of haemophagous regions of placentomes and in other parts of the foetal villous tree, but the strongest immunostaining was in the epithelial cells and lumen of uterine glands. Ultrastructural analyses indicated that erythrophagocytosis was occurring and that erythrocytes were present inside cells of the chorion that also contained endocytic vesicles and caveolae. Results of this study indicate that both the haemophagous areas of placentomes and the areolae at the interface between chorion and endometrial glands are important sites for iron transfer from mother to foetal‐placental tissues in buffalo throughout pregnancy.     

The impacts of uterine environment and fetal genotype on conceptus size and placental vascularity during late gestation in pigs.

Meishan and Yorkshire gilts were bred exclusively to Yorkshire and Meishan boars, respectively, resulting in similar Meishan x Yorkshire fetuses gestating in the uteri of both maternal breeds. Gilts of both breeds were slaughtered on d 90 and 110 of gestation, and the weight and volume of each uterine horn was determined. Fetal weights and crown-rump lengths were recorded. A section of the chorioallantoic-endometrial attachment site was collected for each conceptus and evaluated for placental and endometrial vascular density. An intact placenta was recovered for each conceptus and weighed, and its surface area was determined. Fetal weight and crown-rump length increased (P < .001) markedly between d 90 and 110 of gestation in Meishan and in Yorkshire uteri, but they were markedly less (P < .001) in Meishan than in Yorkshire uteri. Placental weight and placental surface area were reduced by 40% in Meishan, compared with Yorkshire, uteri; however, placental size did not increase between d 90 and 110 in either uterine type. Placental vascular density and associated endometrial vascular density were similar (P > .20) for conceptuses in Meishan and Yorkshire uteri on d 90 and 110 of gestation. Additionally, even though the ratio of fetal weight: placental weight (placental efficiency) increased between d 90 and 110, placental efficiency was similar for conceptuses in Meishan and Yorkshire uteri. In a previous study using straightbred Meishan or Yorkshire conceptuses gestated in either a Meishan or Yorkshire uterus, we found Meishan conceptuses had a markedly greater placental efficiency than did Yorkshire conceptuses, regardless of the type of uterus in which they were gestated. These data indicate that uterine type determines conceptus size and conceptus genotype controls placental efficiency.     

猪胎盘屏障的研究进展

妊娠是一个微妙而复杂的生理过程,胎盘的建立为母胎之间的物质交换奠定了基础。对于猪来说,胎盘功能障碍会降低胎盘效率,引起猪宫内发育迟缓、窝内体重变异增大,给养猪业的经济效益带来巨大损失。胎盘是母胎之间的一道天然屏障,其功能较为复杂,有物质交换、排泄、免疫保护、合成和分泌等多种功能。鉴于胎盘屏障的重要作用,本文综述了猪胎盘屏障的结构、功能及影响因素,以期为相关研究和养猪生产提供参考。     

Conceptus competition for uterine space: different strategies exhibited by the Meishan and Yorkshire pig

Our laboratory has demonstrated that Yorkshire placentae increase in size and surface area during the final third of gestation. In contrast, Meishan placental size remains constant during late gestation, but the density of blood vessels at the placental-endometrial interface increases markedly. Preliminary observations from our laboratory suggest that if one of two adjacent Meishan fetuses dies, the placenta of the remaining Meishan conceptus fails to increase its length of implantation or its placental weight or surface area. In contrast, if one of two adjacent Yorkshire fetuses dies, the adjacent conceptus accelerates its placental growth. The objective of this experiment was to document that Yorkshire, but not Meishan, conceptuses accelerate placental growth when adjacent fetuses are experimentally destroyed on d 40 of gestation. Straightbred Meishan (n = 5) and Yorkshire (n = 5) females were laparotomized and one uterine horn was randomly assigned to receive fetal crushing (treated horn); the other uterine horn served as a within-animal control. In the treated horn, every other fetus was then crushed through the uterine wall and the animals were allowed to recover. On d 111, animals were killed, uteri were recovered, and fetal weight, crown-rump length (CRL), placental weight, implantation site length, and placental surface area were recorded. Although there were no statistically significant differences in fetal weight or CRL observed between treated or control horns of females of either breed, there was a tendency for the fetuses in the treated uterine horn to be longer and heavier in both breeds. There were no differences in placental weight, placental surface area, or implantation site length between conceptuses in Meishan treated and control horns, which averaged 173.8+/-6.4 g, 1,162.7+/-35.9 cm2, and 19.0+/-0.4 cm, respectively. In contrast, placental weight, placental surface area, and implantation site length were increased (P < 0.05) in Yorkshire treated horns compared to Yorkshire control horns (306.1+/-26.0 g, 1,835+/-93.9 cm2, and 33.4+/-1.5 cm vs 253.7+/-13.4 g, 1,474.3+/-50.4 cm2, and 27.2+/-0.8 cm; respectively). These data confirm that Yorkshire conceptuses, but not Meishan conceptuses, accelerate placental growth when adjacent littermates perish as late as d 40 of gestation. These data indicate that differences exist in the strategies employed by Meishan and Yorkshire conceptuses in the competition for nutrients during gestation.     

An approach to comparative anatomy of the acetabulum from amphibians to primates

The objective of this study was to investigate the anatomy, both macroscopic and microscopic, of the soft tissue internal structures of the hip joint in animal species and in three human hips (an adult and two fetuses). We dissected the hip joints of 16 species and compared the anatomical features of the soft tissue from the respective acetabula. In addition, a histological study was made of the specimens studied. In amphibians, we found a meniscus in the acetabulum, which was not observed in any of the other species studied. The isolated round ligament is observed from birds onwards. In the group of mammals analysed, including the human specimens, we found a meniscoid structure in the acetabular hip joint. Furthermore, we found that the meniscoid structure forms an anatomo-functional unit with the round ligament and the transverse ligament of the coxofemoral joint. These discoveries suggest the participation of the soft tissue anatomy in adaptative changes of species.