Morphological characteristics of zebrafish's yolk sac for malformation based on orthogonal-polarization-gating optical coherence tomography.

In this study, an automatic algorithm combining an ellipsoid approximation and U-net has been presented for the characterization of a zebrafish's yolk sac. The polarization-difference-balanced-detection image of zebrafish was obtained based on orthogonal-polarization-gating optical coherence tomography and used to segment the yolk sac region. And ellipsoid can approximate the shape of the three-dimensional yolk sac, and the multiple parameters of volume and the three principal axes (k, l and m) can be used to quantify the yolk sac. In addition, the multiple parameters of two principal axes (l and m) and volume can distinguish the malformation from the normal controlled group. Finally, the volume malformation of the yolk sac calculated by the proposed algorithm ranges from 16.55% to 46.05%. Thus, the degree of malformation can be applied for toxicity analysis. And this method provides a potential application for an accurate judgment index for biotoxicological testing.

Placentation in the Human and Higher Primates.

Placentation in humans is precocious and highly invasive compared to other mammals. Implantation is interstitial, with the conceptus becoming completely embedded within the endometrium towards the end of the second week post-fertilization. Villi initially form over the entire surface of the chorionic sac, stimulated by histotrophic secretions from the endometrial glands. The secondary yolk sac never makes contact with the chorion, and a choriovitelline placenta is never established. However, recent morphological and transcriptomic analyses suggest that the yolk sac plays an important role in the uptake of nutrients from the coelomic fluid. Measurements performed in vivo demonstrate that early development takes place in a physiological, low-oxygen environment that protects against teratogenic free radicals and maintains stem cells in a multipotent state. The maternal arterial circulation to the placenta is only fully established around 10-12 weeks of gestation. By then, villi have regressed over the superficial, abembryonic pole, leaving the definitive discoid placenta, which is of the villous, hemochorial type. Remodeling of the maternal spiral arteries is essential to ensure a high-volume but low-velocity inflow into the mature placenta. Extravillous trophoblast cells migrate from anchoring villi and surround the arteries. Their interactions with maternal immune cells release cytokines and proteases that are key to remodeling, and a successful pregnancy.

Reference intervals of gestational sac, yolk sac, embryonic length, embryonic heart rate at 6-10 weeks after in vitro fertilization-embryo transfer.

BACKGROUND: Accurately determining the normal range of early pregnancy markers can help to predict adverse pregnancy outcomes. The variance in ovulation days leads to uncertain accuracy of reference intervals for natural pregnancies. While the gestational age (GA) is accurate estimation during in vitro fertilization-embryo transfer (IVF-ET). Thus, the objective of this research is to construct reference intervals for gestational sac diameter (GSD), yolk sac diameter (YSD), embryonic length (or crown-rump length, CRL) and embryonic heart rate (HR) at 6-10 gestational weeks (GW) after IVF-ET. METHODS: From January 2010 to December 2016, 30,416 eligible singleton pregnancies were retrospectively recruited. All included participants had full records of early ultrasound measurements and phenotypically normal live neonates after 37 GW, with birth weights > the 5th percentile for gestational age. The curve-fitting method was used to screen the optimal models to predict GSD, CRL, YSD and HR based on gestational days (GD) and GW. Additionally, the percentile method was used to calculate the 5th, 50th, and 95th percentiles. RESULTS: There were significant associations among GSD, CRL, YSD, HR and GD and GW, the models were GSD = - 29.180 + 1.070 GD (coefficient of determination [R2] = 0.796), CRL = - 11.960 - 0.147 GD + 0.011 GD2 (R2 = 0.976), YSD = - 2.304 + 0.184 GD - 0.011 GD2 (R2 = 0.500), HR = - 350.410 + 15.398 GD - 0.112 GD2 (R2 = 0.911); and GSD = - 29.180 + 7.492 GW (R2 = 0.796), CRL = - 11.960 - 1.028 GW + 0.535 GW2 (R2 = 0.976), YSD = - 2.304 + 1.288 GW - 0.054 GW2 (R2 = 0.500), HR = - 350.410 + 107.788 GW - 5.488 GW2 (R2 = 0.911), (p < 0.001). CONCLUSIONS: Reference intervals for GSD, YSD, HR and CRL at 6-10 gestational weeks after IVF-ET were established.

Early pregnancy ultrasound measurements and prediction of first trimester pregnancy loss: A logistic model.

Our objective was to prospectively validate the use of gestational sac (GS), yolk sac (YS) diameter, crown-rump length (CRL), and embryonal heart rate (HR) dimensions to identify early pregnancy loss. This was a prospective cohort study of first trimester pregnancies. GS and YS diameter, CRL, and HR measurements were serially obtained in singleton and twin pregnancies from 6 through 10 weeks' gestation. Non-parametric tests and logistic regression models were used for comparisons of distributions and testing of associations. A total of 252 patients were included, of which 199 were singleton pregnancies, 51 were twins, and 2 were triplets (304 total fetuses). Fifty-two patients had 61 losses. We built nomograms with the changes of the parameters evaluated in ongoing, as well as in pregnancy loss. In the pregnancies which failed, all the parameters showed significant changes, with different temporal onsets: GS and YS were the first to become abnormal, deviating from normality as early as 6 weeks' gestation (OR 0.01, 95% CI 0.0-0.09, and OR 3.36, 95% CI 1.53-7.34, respectively), followed by changes in HR, and CRL, which became evident at 7 and 8 weeks (OR 0.96, 95% CI 0.92-1.0, and OR 0.59, 95% CI 0.48-0.73, respectively). Our observations showed that, after 5 complete weeks' gestation, a small GS and a large YS reliably predicted pregnancy loss. The YS reliably identified the occurrence of a miscarriage at least 7 days prior its occurrence. CRL and HR became abnormal at a later time in pregnancy and closer to the event. These findings have important implications for patient counseling and care planning, as well as a potential bearing on cost effectiveness within early pregnancy care.

Novel placental structure in the Mexican gerrhonotine lizard, Mesaspis viridiflava (Lacertilia; Anguidae).

The evolution of viviparity alters the physical relationship between mothers and offspring and the prevalence of viviparity among squamate reptiles presents an opportunity to uncover patterns in the evolution of placental structure. Understanding the breadth of this diversity is limited because studies of placental structure and function have emphasized a limited number of lineages. We studied placental ontogeny using light microscopy for an embryological series of the Mexican gerrhonotine lizard, Mesaspis viridiflava. This species develops an elaborate yolk sac placenta, an omphaloplacenta, which receives vascular support arising in a structure known only from other gerrhonotine lizards. A prominent feature of the omphaloplacenta is a zone of uterine and embryonic epithelial cell hyperplasia located at the upper shoulder of the yolk mass, often extending above the yolk mass. The omphaloplacenta covers more than one-half of the surface area of maternal-embryonic contact. The chorioallantoic placenta has a more restricted distribution because the allantois remains in the embryonic hemisphere of the egg throughout development and lies internal to the vascular support for the omphaloplacenta in areas where they overlap. The structural profile of the chorioallantoic placenta indicates a potential for respiratory exchange and/or hemotrophic nutritive transport, while that of the omphaloplacenta suggests that nutritive transfer is primarily via histotrophy. An eggshell is present in the earliest embryonic stages examined but regresses relatively early in development. Placental specializations of this species are consistent with a pattern of matrotrophic embryonic nutrition and have evolved in a unique lineage specific developmental pattern.

Yolk sac size & shape as predictors of first trimester pregnancy outcome: A prospective observational study.

OBJECTIVE: To determine the value of yolk sac size and shape for prediction of pregnancy outcome in the first trimester. MATERIAL AND METHODS: 500 pregnant women between 6+0 and 9+6 weeks of gestation underwent transvaginal ultrasound and yolk sac diameter (YSD), gestational sac diameter (GSD) were measured, presence/absence of yolk sac (YS) and shape of the yolk sac were noted. Follow up ultrasound was done to confirm fetal well-being between 11+0 and 12+6 weeks and was the cutoff point of success of pregnancy. RESULTS: Out of 500 cases, 8 were lost to follow up, YS was absent in 14, of which 8 were anembryonic pregnancies. Thus, 478 out of 492 followed up cases were analyzed for YS shape and size and association with the pregnancy outcome. In our study, abnormal yolk sac shape had a sensitivity and specificity (87.06% & 86.5% respectively, positive predictive value (PPV) of 58.2%, negative predictive value (NPV) of 96.8% in predicting a poor pregnancy outcome as compared to yolk sac diameter (sensitivity and specificity 62.3% & 64.1% respectively and PPV and NPV of 27.3% and 88.7% respectively). The degree of association for both the variables was significant to the level of p<0.000. CONCLUSION: The presence or absence of yolk sac has a strong predictive value for poor pregnancy outcome. Yolk sac shape was a better predictor of poor pregnancy outcome in terms of higher specificity and negative predictive value as compared to yolk sac diameter.

Yolk sac-embryo distance in correlation with soluble form of vascular endothelial growth factor levels in pregnancy with potentially reserved evolutivity.

AIMS: Embryonic demise is a frequent complication of the first trimester pregnancy. The purpose of this study was to evaluate the correlation between a serum biomarker, the soluble form of the vasculo-endothelial growth factor (sFlt-1) and the distance between the yolk sac (YS) and embryo (DYSE), determined by ultrasonography. MATERIAL AND METHODS: The study was a prospective case-control study that included 2 groups of patients - the control group with 81 first-trimester pregnancies in evolution and the case group with 89 first-trimester pregnancies with a potentially reserved evolutivity. RESULTS: A correlation between the serum level of sFlt-1 and DYSE in embryos with crown-rump length (CRL) greater than 5 mm was identified, showing that a DYSE ≤3 mm correlates with a low level of sFlt-1 (p<0.05) and a DYSE> 4 mm correlates with an increased level of sFlt-1 (p<0.05). CONCLUSIONS: A low level of sFlt-1 associated with a distance between the embryo and yolk sac of small dimensions, respectively <3 mm, correlates with an increased rate of non-viable embryos. This correlation between an ultrasound and a serum parameter is of great value and brings important information about the viability of firsttrimester pregnancies.

Environmental calcium and variation in yolk sac size influence swimming performance in larval lake sturgeon (Acipenser fulvescens).

In many animal species, performance in the early life stages strongly affects recruitment to the adult population; however, factors that influence early life history stages are often the least understood. This is particularly relevant for lake sturgeon, Acipenser fulvescens, living in areas where environmental calcium concentrations are declining, partly due to anthropogenic activity. As calcium is important for muscle contraction and fatigue resistance, declining calcium levels could constrain swimming performance. Similarly, swimming performance could be influenced by variation in yolk sac volume, because the yolk sac is likely to affect drag forces during swimming. Testing swimming performance of larval A. fulvescens reared in four different calcium treatments spanning the range of 4-132 mg l-1 [Ca2+], this study found no treatment effects on the sprint swimming speed. A novel test of volitional swimming performance, however, revealed reduced swimming performance in the low calcium environment. Specifically, volitionally swimming larvae covered a shorter distance before swimming cessation in the low calcium environment compared with the other treatments. Moreover, sprint swimming speed in larvae with a large yolk sac was significantly slower than in larvae with a small yolk sac, regardless of body length variation. Thus, elevated maternal allocation (i.e. more yolk) was associated with reduced swimming performance. Data suggest that larvae in low calcium environments or with a large yolk sac exhibit reduced swimming performance and could be more susceptible to predation or premature downstream drift. Our study reveals how environmental factors and phenotypic variation influence locomotor performance in a larval fish.

The yolk syncytial layer of loach, Misgurnus fossilis (Teleostei) during early development.

The yolk syncytial layer (YSL) of Teleostei is a dynamic multifunctional temporary system. This paper describes the YSL structure of Misgurnus fossilis (Cobitidae) during its early developmental stages, studied using histological methods. YSL formation is prolonged. From the late blastula stage, the basal surface of the YSL is uneven and has protuberances, but becomes smoother during development. There are syncytial 'islands' with 1-2 yolk syncytial nuclei in the yolk mass. During epiboly, gastrulation and early segmentation, loach YSL is of different thickness in different regions along the dorso-ventral and antero-posterior axes of an embryo. The YSL is thickened in the dorsal region of gastrulae compared with the ventral region. Although the development of M. fossilis is similar to the development of zebrafish, there are important differences in YSL formation and organization that await further study and analysis. The study of YSL organization contributes to our knowledge of teleost developmental diversity and to the biology of temporary structures.

Morphological specializations of the yolk sac for yolk processing in embryonic corn snakes (Pantherophis guttatus: Colubridae).

Non-avian reptiles commonly are assumed to be like birds in their overall patterns of development. However, colubrid corn snakes (Pantherophis guttatus) have mechanisms of yolk cellularization and processing that are entirely different from the avian pattern. In birds, a vascular "yolk sac" surrounds and digests the liquid yolk. In contrast, in corn snakes, the yolk material is converted into vascularized cords of yolk-filled cells. In this study, we used stereomicroscopy, histology, and scanning electron microscopy to analyze this unusual developmental pattern in corn snakes. Our observations reveal that the yolk sac cavity is invaded by endodermal cells that proliferate, absorb yolk spheres, and form aggregates of interconnected cells within the liquid yolk mass. As development proceeds, small blood vessels arise from the yolk sac omphalopleure, penetrate into the yolk mass, and become tightly encased in the endodermal cells. The entire vitellus ultimately becomes converted into a mass of vascularized, "spaghetti-like" strands of yolk-laden cells. The resulting arrangement allows yolk to be digested intracellularly and yolk products to be transported to the developing embryo. Indirect evidence for this pattern in other species raises the possibility that it is ancestral for squamates and quite possibly Reptilia in general.

Morphology, histochemistry and glycosylation of the placenta and associated tissues in the European hedgehog (Erinaceus europaeus).

INTRODUCTION: There are few descriptions of the placenta and associated tissues of the European hedgehog (Erinaceus europaeus) and here we present findings on a near-term pregnant specimen. METHODS: Tissues were examined grossly and then formalin fixed and wax-embedded for histology and immunocytochemistry (cytokeratin) and resin embedded for lectin histochemistry. RESULTS: Each of four well-developed and near term hoglets displayed a discoid, haemochorial placenta with typical labyrinth and spongy zones. In addition there was a paraplacenta incorporating Reichert's membrane and a largely detached yolk sac. The trophoblast of the placenta contained diverse populations of granule which expressed most classes of glycan. Intercellular membranes were also glycosylated and this tended to be heavier in the labyrinth zone. Fetal capillary endothelium had glycosylated apical surfaces expressing sialic acid and various other glycans. Glycogen was present in large cells situated between the spongy zone and the endometrium. Trophoblast cells in the placental disc and under Reichert's membrane, as well as yolk sac endoderm and mesothelium, were cytokeratin positive. Reichert's membrane was heavily glycosylated. Yolk sac inner and outer endoderm expressed similar glycans except for N-acetylgalactosamine residues in endodermal acini. DISCUSSION: New features of near-term hedgehog placenta and associated tissues are presented, including their glycosylation, and novel yolk sac acinar structures are described. The trophoblast of the placental disc showed significant differences from that underlying Reichert's membrane while the glycan composition of the membrane itself showed some similarity to that of rat thereby implying a degree of biochemical conservation of this structure.

The influence of heart developmental anatomy on cardiotoxicity-based adverse outcome pathways in fish.

The developing fish heart is vulnerable to a diverse array of toxic chemical contaminants in freshwater, estuarine, and marine habitats. Globally occurring examples of cardiotoxic agents include dioxins, polychlorinated biphenyls (PCBs), and polycyclic aromatic hydrocarbons (PAHs). The disruption of cardiac function during the process of heart morphogenesis can lead to adverse outcome pathways (AOPs) that can negatively affect fish survival at hatching as well as later life stages. Proximal impacts include cardiogenic fluid accumulation (edema) and defects of the body axis and jaw that preclude larval feeding. More subtle changes in heart development can produce permanent structural defects in the heart that reduce cardiac output and swimming performance in older fish. In recent decades, the presence of edema in fish embryos and larvae has been a very common bioindicator of cardiotoxicity. However, the different ways that edema forms in fish from different habitats (i.e., freshwater vs. marine, pelagic vs. demersal) has not been rigorously examined. Oil spills are an important source of PAHs in fish spawning areas worldwide, and research is revealing how patterns of cardiogenic edema are shaped by species-specific differences in developmental anatomy and ionoregulatory physiology. Here we review the visible evidence for circulatory disruption across nine freshwater and marine fish species, exposed to crude oils from different parts of the world. We focus on the close interconnectedness of the cardiovascular and osmoregulatory systems during early development, and corresponding implications for fish in hyperosmotic and hyposmotic habitats. Finally, we suggest there may be poorly understood adverse outcomes pathways related to osmotic gradients and water movement within embryos, the latter causing extreme shifts in tissue osmolality.

Histomorphological changes in digestive tract of golden mahseer (Tor putitora) during different developmental stages.

Histomorphological changes in digestive tract of golden mahseer (Tor putitora) were examined in larvae [starting from hatching to 45 days post-hatching (dph)], fry, fingerling, and adult. Digestive tract appeared during hatching, on the dorsal side of yolk sac, as a straight tube with a narrow lumen. Mouth opening and appearance of liver and pancreas were observed at 2 dph, and subsequently anal opening, appearance of goblet cells in esophagus, and posterior intestine were evident at 3 dph. The remodeling of oral cavity in terms of epithelial stratification, appearance of taste buds, and goblet cells were observed in a window of 4-5 dph. Intestinal folding was found to be initiated at 8 dph. From 12 to 45 dph, thickening of oral and esophageal mucosal/extramucosal layers, increase in intestinal folding, increases in the density of goblet cells in entire gut were observed. Within the same time window, other histological changes such as disappearance of vacuoles in liver, and abundance of zymogen granules in pancreas were also observed. Supranuclear vesicles in mid-to-posterior intestine were found to be prominent from first feeding to 45 dph; however, this phenomenon was no longer evident in fry and fingerling. Overall, the increase in intestinal folding and complexity of extramucosal layer were found to be continuous from the first appearance to adult, and this inferred the fact that the nutritional physiology, in terms of digestion and assimilation, progressively changes throughout the life stages of golden mahseer. Findings of this study will, therefore, help in preparing diets for different life stages of this fish, and in addition, the present information widens the understanding of digestive physiology of golden mahseer.

Determination of limited histotrophy as the reproductive mode in Mustelus schmitti Springer, 1939 (Chondrichthyes: Triakidae): analysis of intrauterine growth of embryos

This work analyzed length-mass relationship, growth in length, mass variations and dynamic of yolk sac depletion of Mustelus schmitti embryos, extracted from females catched by the artisanal fishery acting off La Paloma (34°39'S, 54°10'W, Uruguay) during 2006 and 2007. Differences in total length and total mass between sexes were not significant. Embryos showed a negative allometric growth for both years with a slope change near the day 170 of the year, likely to be associated with the depletion of the external yolk sac. Gompertz and Von Bertalanffy curves were adjusted for each year. Parturition date was determined around the day 300 of the year. Yolk depletion followed a logistic dynamic Yt=1/(1+e(-13.749 + 0.072*t)) for 2006 and Yt=1/(1+e(-10.472 + 0.054*t)) for 2007. Embryos showed a mean increase in total dry mass of 5.4g in 187 days which, together with yolk depletion dynamics, indicates additional energetic supply, suggesting that M. schmitti presents limited histotrophy.

Este trabajo analizó las relaciones largo-masa, crecimiento en talla, variaciones en masa y dinámica de consumo del saco vitelino de embriones de Mustelus schmitti extraídos de hembras capturadas por la flota pesquera artesanal del puerto de La Paloma (34°39'S, 54°10'W, Uruguay) durante 2006 y 2007. Las diferencias en largo total y masa total entre sexos no fueron significativas. Los embriones mostraron crecimiento alométrico negativo en ambos años, con un cambio de pendiente cercano al día 170, probablemente asociado con el agotamiento del saco vitelino externo. Se ajustaron curvas de crecimiento de Gompertz y VonBertalanffy para cada año. La fecha de parición se determinó cerca del día 300 del año. El consumo de vitelo siguió una dinámica logística Yt=1/(1+e(-13.749 + 0.072*t)) en 2006 y Yt=1/(1+e(-10.472 + 0.054*t)) en 2007. Los embriones mostraron un aumento promedio de masa seca de 5.4g en 187 días, esto, junto con la dinámica de consumo de vitelo, indicó una provisión adicional de energía, sugiriendo que M. schmitti presenta histotrofía limitada.

Ultrastructure of the fetal membranes of the oviparous kingsnake, Lampropeltis getula (Colubridae) as revealed by scanning electron microscopy.

In reptilian sauropsids, fetal (extraembryonic) membranes that line the eggshell sustain developing embryos by providing for gas exchange and uptake of water and eggshell calcium. However, a scarcity of morphological studies hinders an understanding of functional specializations and their evolution. In kingsnakes (Lampropeltis getula), scanning electron microscopy reveals two major fetal membranes: the chorioallantois and yolk sac omphalopleure. In early development, the chorioallantois contains tall chorionic epithelial cells, avascular connective tissue, and enlarged allantoic epithelial cells. During its maturation, the chorionic and allantoic epithelia thin dramatically and become underlain by a rich network of allantoic capillaries, yielding a membrane ideally suited for respiratory gas exchange. Yolk sac development initially is like that of typical lizards and snakes, forming an avascular omphalopleure, isolated yolk mass (IYM), and yolk cleft. However, unlike the situation in most squamates studied, the omphalopleure becomes transformed into a "secondary chorioallantois" via three asynchronous events: flattening of the epithelium, regression of the IYM, and vascularization by the allantois. Progressive expansion of chorioallantois parallels growing embryonic needs for gas exchange. In early through mid-development, external surfaces of both the chorionic and omphalopleure epithelium show an abundance of irregular surface protrusions that possibly increase surface area for water absorption. We postulate that the hypertrophied allantoic epithelial cells produce allantoic fluid, a viscous substance that facilitates water uptake and storage. Our findings are consistent with a previous study on the corn snake Pantherophis guttatus, but include new observations and novel functional hypotheses relevant to a reconstruction of basal squamate patterns.

Dinâmica da inversão do saco vitelino em preás (Galea spixii Wagler, 1831) / Dynamics of yolk sac inversion in galea (Galea spixii Wagler, 1831)

O objetivo deste trabalho foi estudar o período de inversão do saco vitelino bem como a dinâmica resultante deste processo na gestação inicial em preás, utilizando-se microscopia de luz, microscopia eletrônica de varredura e de transmissão. No décimo segundo dia de gestação observou-se o desenvolvimento dos endodermas parietal e visceral delimitando a cavidade do saco vitelino. O endoderma parietal foi evidenciado revestindo a superfície fetal da placenta corioalantoidea bem como contornando o espaço delimitado pela decídua capsular. Estes endodermas apresentaram formato prismático e encontraram-se separados do trofoblasto por uma desenvolvida membrana de Reichert. Já o endoderma visceral continha vasos vitelínicos e possuía vilosidades apenas em determinadas áreas. No décimo quarto dia de gestação verificou-se a inversão do saco vitelino, caracterizada pela degeneração do endoderma parietal e trofoblasto mural, associado ao desaparecimento gradual da membrana de Reichert. Como consequência deste fenômeno, o endoderma visceral passou a constituir uma interface com o epitélio uterino. Após a inversão, o endoderma parietal que permaneceu íntegro foi aquele que se apoiava na superfície da placenta corioalantóidea, apresentando células em formato colunar alto e característica de epitélio pseudoestratificado. O endoderma visceral apresentou numerosas vilosidades apicais principalmente em regiões próximas a placenta corioalantóidea. Com o contínuo desenvolvimento do embrião e placenta corioalantóidea, observou-se o surgimento de importante área de aposição entre os endodermas visceral e parietal. A inversão do saco vitelino representou uma disposição anatômica favorável ao desenvolvimento embrionário, além de ser uma característica evolutiva nesta espécie de roedor.

The aim of this study was to study the time of yolk sac inversion as well as the dynamics resulting from this process in galea throughout pregnancy. For this, conventional histological techniques, scanning electron microscopy and transmission electron microscopy were used. Parietal and visceral endoderm delimiting the yolk sac cavity was observed at 12 days of pregnancy. The parietal endoderm was coating the fetal surface of the chorioallantoic placenta as well as delimiting the decidua capsularis area. This endoderm had prismatic format and were apart from the trophoblast by an enlarged Reichert's membrane. The visceral endoderm had vitelline vessels and there were villi only in certain areas. At 14 days of pregnancy the yolk sac inversion was characterized by the degeneration of parietal endoderm and mural trophoblast, and also the gradual disappearance of the Reichert's membrane. So it made the visceral endoderm establish an interface with the uterine epithelium. After the inversion, the parietal endoderm which remained intact was the one that rested on the chorioallantoic placenta surface. It presented cells with high columnar format and pseudostratified epithelium featured. The visceral endoderm presented many apical villi, especially in areas close to the chorioallantoic placenta. The continued development of the embryo and chorioallantoic placenta evidenced the emergence of an important apposition area between visceral and parietal endoderm. The yolk sac inversion represented an anatomical arrangement in favor of the embryo development as well as an evolutionary trait in this rodent species.

Development and morphology of the inverted yolk sac in the guinea pig (Cavia porcellus).

Although the guinea pig is an important animal model for human placentation, aspects of fetal nutrition are not fully understood, especially in regard to the yolk sac that is regarded to be essential for early development of the embryo. We investigated differentiation by means of histology, histochemistry, immunohistochemistry, and transmission electron microscopy. Data suggest that the guinea pig's yolk sac was not sufficiently developed to facilitate substantial fetal nutrition in early pregnancy. On Day 12, it was a flat, inverted, but avascular structure. This was followed by differentiation to form the typical, highly villous and vascularized condition of advanced gestation. Finally, the yolk sac degenerated toward term. We suggest that the guinea pig and other caviomorphs rely predominantly on hemotrophic nutrition via the placenta even in very early pregnancy. In contrast to the general pattern of mammals, histiotrophic nutrition via yolk sac routes seems to be most essential during mid-gestation.

Yolk sac endoderm is the major source of serum proteins and lipids and is involved in the regulation of vascular integrity in early chick development.

An important function of the vascular system is nutrient delivery. In adult animals, this is mediated through a close contact of the mesoderm-derived vasculature with the endoderm-derived enterocytes and hepatocytes. During embryonic development, the yolk sac (YS) endoderm has been suggested to play a similar role. Physiological and molecular nature of the contact between the YS endoderm and the vasculature is not well-understood. To understand roles of the YS endoderm in early development, we used the avian model and carried out a gene expression profiling analysis of isolated area vasculosa YS endoderm tissues from embryonic day 2-4 chick embryos, covering the first 48 hr of postcirculation development. Genes involved in lipid metabolism are highly enriched, indicating an active modification of lipid components during their transfer from the yolk to the circulatory system. We also uncovered genes encoding major serum proteins and key regulators of vascular integrity. In particular, PTGDS, an enzyme controlling the last step of prostaglandin D2 production, shows high expression in the YS endoderm. Experimental introduction of prostaglandin D2 into embryonic circulation led to intraembryonic vessel rupture. These data suggest that the YS endoderm is the major, if not exclusive, source of lipid and protein constituents of the early embryonic serum and plays an important role in the regulation of vascular integrity in developing embryo.

Mesothelium of the murine allantois exhibits distinct regional properties.

The rodent allantois is thought to be unique amongst mammals in not having an endodermal component. Here, we have investigated the mesothelium, or outer surface, of murine umbilical precursor tissue, the allantois (∼7.25-8.5 days postcoitum, dpc) to discover whether it exhibits the properties of an epithelium. A combination of morphology, challenge with biotinylated dextran amines (BDAs), and immunohistochemistry revealed that the mesothelium of the mouse allantois exhibits distinct regional properties. By headfold stages (∼7.75-8.0 dpc), distal mesothelium was generally squamous in shape, and highly permeable to BDA challenge, whereas ventral proximal mesothelium, referred to as "ventral cuboidal mesothelium" (VCM) for the characteristic cuboidal shape of its cells, was relatively impermeable. Although "dorsal cuboidal mesothelium" (DCM) resembled the VCM in cell shape, its permeability to BDA was intermediate between the other two regions. Results of immunostaining for Zonula Occludens-1 (ZO-1) and Epithelial-cadherin (E-cadherin), together with transmission electron microscopy (TEM), suggested that impermeability in the VCM may be due to greater cellular contact area between cells and close packing rather than to maturity of tight junctions, the latter of which, by comparison with the visceral yolk sac, appeared to be rare or absent from the allantoic surface. Both VCM and DCM exhibited an ultrastructure more favorable for protein synthesis than did the distal squamous mesothelium; however, at most stages, VCM exhibited robust afadin (AF-6), whereas the DCM uniquely contained alpha-4-integrin. These observations demonstrate that the allantoic mesothelium is not a conventional epithelium but possesses regional ultrastructural, functional and molecular differences that may play important roles in the correct deployment of the umbilical cord and its associated vascular, hematopoietic, and other cell types.

Scanning electron microscopy of the placental interface in the viviparous lizard Sceloporus jarrovi (Squamata: Phrynosomatidae).

Placental membranes mediate maternal-fetal exchange in all viviparous reptilian sauropsids. We used scanning electron microscopy to examine the placental interface in the mountain spiny lizard, Sceloporus jarrovi (Phrynosomatidae). From the late limb bud stage until birth, the conceptus is surrounded by placental membranes formed from the chorioallantois and yolk sac omphalopleure. The chorioallantois lies directly apposed to the uterine lining with no intervening shell membrane. Both fetal and maternal sides of the chorioallantoic placenta are lined by continuous layers of flattened epithelial cells that overlie dense capillary networks. The chorioallantoic placenta shows specializations that enhance respiratory exchange, as well as ultrastructural evidence of maternal secretion and fetal absorption. The yolk sac placenta contains enlarged fetal and maternal epithelia with specializations for histotrophic nutrient transfer. This placenta lacks intrinsic vascularity, although the vascular allantois lies against its inner face, contributing to an omphallantoic placenta. In a specialized region at the abembryonic pole, uterine and fetal tissues are separated by a compact mass of shed shell membrane, yolk droplets, and cellular debris. The omphalopleure in this region develops elongate folds that may contribute to sequestration and absorption of this material. Fetal membrane morphogenesis and composition in S. jarrovi are consistent with those of typical squamates. However, this species exhibits unusual placental specializations characteristic of highly placentotrophic lizards.