In ovo injection of testosterone to yolk sac modulates early posthatching development and physiology of male chick in broilers.

The aim of this study was to investigate the effects of in ovo testosterone injection into the yolk sac of embryos on physiology and development of broiler chicks during the early posthatching period. A total of 1,010 hatching eggs were obtained from the Ross genotype. Trial design was conducted with a noninjected group (control) and injection groups in which 100 µL sesame oil, or 100 µL sesame oil + 0.50 µmol testosterone were injected into the yolk sac of the embryo on d 6 or d 12 of incubation. Testosterone hormone level was measured in the egg yolk and albumen at onset of incubation, in the yolk sac on d 19 of incubation and in the residual yolk sac at hatching. Weights of chick, yolk sac and organ, morphological traits (body length, lengths of bilateral traits and beak length), asymmetrical development of bilateral morphological traits and body mass index were measured at hatching and on d 7 after hatching. Testosterone, corticosterone and growth hormone levels were determined in blood plasma obtained from male chicks at hatching and on d 7 of chick age. Chick weight was not affected, plasma testosterone level and brain weight decreased, while body mass index, plasma corticosterone and growth hormone levels increased by administering 0.50 µmol testosterone on d 12 of embryonic age. However, plasma testosterone and growth hormone levels did not change, chick weight increased, while plasma corticosterone level and the chick body length decreased by administering 0.50 µmol testosterone on d 6 of embryonic age. A significant interaction between chick age and in ovo testosterone administration resulted in an increase in lung weight of chicks. In conclusion, this study found that in ovo testosterone administered at different embryonic ages due to age-specific effects of testosterone in the yolk sac of embryo modulates development related to physiological parameters of male broiler chicks during early posthatching period.

Influence of the preplacement holding time and feeding hydration supplementation before placement on yolk sac utilization, the crop filling rate, feeding behavior and first-week broiler performance.

This study investigated the effects of the broiler chick preplacement holding time and feeding hydration supplementation before placement on yolk sac utilization, the crop filling rate, feeding-drinking behavior and first-wk broiler performance. Broiler hatching eggs were obtained from a commercial broiler breeder flock of Ross 308 at 37 wk of age and incubated in a commercial hatchery. At 510 h of incubation, all chicks were removed from the hatcher and separated into cardboard chick boxes containing 80 chicks each. The chick boxes were randomly separated into two groups with either added commercial hydration supplementation (gel: Hydrogel-95) or the control (no gel). Then, the chicks were randomly distributed into 5 groups with different holding times across each hydration supplementation treatment (gel and control). The preplacement holding times were 6, 24, 48, 60, and 72 h from the pull time from the hatchers in the hatchery to placement in the broiler house on the farm, at which point the chicks were able to access feed and water. There were 10 subtreatment groups comprising 5 chick preplacement holding time groups × 2 hydration supplementation groups. There were 12 replicates (160 chicks per pen) per holding period × gel treatment, with a total of 19,200 chicks placed. The feed and water access time did not influence yolk sac utilization, but the absolute or relative residual yolk sac (g, %) decreased linearly with the duration after the pull time (P < 0.001). Longer preplacement holding times were associated with a higher percentage of chicks with full crops at 3 h after placement (P < 0.001). Chicks with the shortest (6 h) preplacement holding time had a lower percentage of feed-seeking activity compared to the 24, 48, and 72 h holding time groups at 3 h after placement (P < 0.001). The highest chick eating and drinking activity was observed in the 72 h group at both 3 and 8 h after placement. Chick weight at placement was significantly reduced linearly with the duration after the pull time (0.106 g/h; R2 = 0.775), and as expected, the highest and lowest BW were found in the 6 (41.51 g) and 72 h (34.50 g) preplacement holding time groups, respectively. However, BW and BW gain were higher in the 24 h group than in the other preplacement holding time groups (P < 0.001) at 7 d after placement. Mortality within the first 3 d after placement increased only when the preplacement holding time was extended to 72 h (P = 0.002). Mortality during 4 to 7 d postplacement was not affected by the holding time at all, but the 72-h holding time group still had statistically significantly higher mortality cumulatively from 0 to 7 d (P = 0.024). Neither BW nor mortality was affected by feeding the hydration supplement at placement, and the lack of effect persisted through 7 d after placement (P > 0.05). It can be concluded that the BW at 7 d after placement was greater in the 24 h holding time group than in shorter (6 h) or longer (48, 60, and 72 h) preplacement holding time groups. In the present study, a greater number of chicks were raised, and it was clearly demonstrated that mortality, as a direct indicator of flock health and welfare, was not affected by preplacement holding times up to and including a 60 h after take-off under thermal comfort conditions, but holding for a further 12 h to 72 h, mortality at 7 d of age after placement was increased. On the other hand, holding chicks in a short period (6 h) did not improve mortality and the BW at 7 d, suggesting that some delay to placement can be beneficial. In addition, feeding hydrogel during the preplacement holding period had no positive effect on BW gain and cumulative mortality during the first week of the growing period.

The effect of an isoquinoline alkaloid on treatment of periodontitis by regulating the neutrophils chemotaxis.

Neutrophil plays a critical role in the progression of periodontitis. In general, its chemotaxis and activation are benefit for the host defense of bacterial infection and inflammation. However, previous studies have reported that the hyperactive and reactive neutrophils appear to be one of the reasons for tissue destruction in periodontitis tissues. In this study, we investigated an isoquinoline alkaloid Litcubanine A (LA), which from the Traditional Chinese medicinal plant, Litsea cubeba. We found LA showed significant activity in inhibiting neutrophils chemotaxis in the zebrafish yolk sac microinjection model in vivo and in mouse neutrophils in vitro. Further investigation proved that LA could inhibit the expression levels of neutrophil respiratory burst-related and inflammation-related genes CYBB and NCF2, as well as inhibit the activation of MAPK signaling pathway. Moreover, using LA, we successfully achieved the effect of reducing periodontitis bone loss by regulating neutrophil chemotaxis and related functions in a mouse ligature-induced periodontitis model.

Maternal Inositol Status and Neural Tube Defects: A Role for the Human Yolk Sac in Embryonic Inositol Delivery?

Supplementation with myo-inositol during the periconceptional period of pregnancy may ameliorate the recurrence risk of having a fetus affected by a neural tube defect (NTD; e.g., spina bifida). This could be of particular importance in providing a means for preventing NTDs that are unresponsive to folic acid. This review highlights the characteristics of inositol and describes the role of myo-inositol in the prevention of NTDs in rodent studies and the evidence for its efficacy in reducing NTD risk in human pregnancy. The possible reduction in NTD risk by maternal myo-inositol implies functional and developmentally important maternal-embryonic inositol interrelationships and also suggests that embryonic uptake of myo-inositol is crucial for embryonic development. The establishment of active myo-inositol cellular uptake mechanisms in the embryonic stages of human pregnancy, when the neural tube is closing, is likely to be an important determinant of normal development. We draw attention to the generation of materno-fetal inositol concentration gradients and relationships, and outline a transport pathway by which myo-inositol may be delivered to the early developing human embryo. These considerations provide novel insights into the mechanisms that may underpin inositol's ability to confer embryonic developmental benefit.

Optimal combination of anti-inflammatory components from Chinese medicinal formula Liang-Ge-San.

ETHNOPHARMACOLOGICAL RELEVANCE: Liang-Ge-San (LGS), a traditional Chinese medicine (TCM) formula, is usually used in acute inflammatory diseases in China. AIM OF THE STUDY: This study aims to detect the optimal combination of anti-inflammatory components from LGS. MATERIALS AND METHODS: Four mainly representative components (phillyrin, emodin, baicalin, and liquiritin) from LGS were chosen. The optimal combination was investigated by orthogonal design study. Zebrafish inflammation model was established by lipopolysaccharide (LPS)-yolk microinjection, and then the anti-inflammatory activities of different combinations were determined by survival analysis, changes on inflammatory cells infiltration, the MyD88/NF-κB and MAPK pathways and inflammatory cytokines production. RESULTS: The different combinations of bioactive ingredients from LGS significantly protected zebrafish from LPS-induced inflammation, as evidenced by decreased recruitment of macrophages and neutrophils, inhibition of the MyD88/NF-κB and MAPK pathways and down-regulation of TNF-α and IL-6. Among them, the combination group 8 most significantly protected against LPS. The combination of group 8 is: 0.1 µM of emodin, 2 µM of baicalin, 20 µM of phillyrin and 12.5 µM of liquiritin. CONCLUSION: The optimized combination group 8 exerts the most significant anti-inflammatory activity by inhibiting the recruitment of inflammatory cells, activation of the MyD88/NF-κB and MAPK pathways and the secretion of pro-inflammatory cytokines. This present study provides pharmacological evidences for the further development of new modern Chinese drug from LGS to treat acute inflammatory diseases, but indicated the use of zebrafish in the screening of components from formulas.

Effects of maternal dietary vitamin E on the egg characteristics, hatchability and offspring quality of prolonged storage eggs of broiler breeder hens.

This research aims to evaluate the effects of maternal vitamin E (VE) dietary supplementation on the egg characteristics, hatchability and antioxidant status of the embryo and newly hatched chicks of prolonged storage eggs. A total of 576 75-week-old Ross 308 breeder hens were randomly allocated into three dietary VE treatments (100, 200 and 400 mg/kg) with 6 replicates of 32 hens, for a 12-week feeding trial. At week 12, a total of 710 eggs were collected over a 5-day period, and eggs per treatment were attributed into 5 replicates and stored for 14 days until incubation. The egg yolk, trunk and head of 7-day-old embryo and the serum, liver, brain and yolk sac of newly hatched chicks were sampled for the evaluation of antioxidant status. Results showed that as maternal dietary VE levels increased, yolk α-tocopherol concentration increased (p < .05). Compared with 100 mg/kg VE, the use of 200 and 400 mg/kg VE increased the hatchability of set/fertile eggs and total antioxidant capacity (T-AOC) of liver and serum in chicks (p < .05), and decreased both the early embryonic mortality and the malondialdehyde (MDA) content of trunk and head in 7-day-old embryos (p < .05); moreover, 400 mg/kg VE increased the yolk T-AOC (p < .05) and decreased yolk and brain MDA content of chicks (p < .05). Brain T-AOC of chicks in 200 mg/kg VE group was improved compared to that of chicks in 100 mg/kg VE group (p < .05). In conclusion, maternal dietary VE at 200 or 400 mg/kg could increase hatchability by decreasing early embryonic mortality and increasing the antioxidant status of egg yolk, embryo and newly hatched chicks as breeder egg storage was prolonged to 14-18 days. The suitable VE level for the broiler breeder diet was 200 mg/kg as breeder egg storage was prolonged.

Porous Yolk-Shell Particle Engineering via Nonsolvent-Assisted Trineedle Coaxial Electrospraying for Burn-Related Wound Healing.

Yolk-shell particles (YSPs) have attracted increasing attention from various research fields because of their low density, large surface area, and excellent loading capacity. However, the fabrication of polymer-based porous YSPs remains a great challenge. In this work, multifunctional polycaprolactone YSPs were produced using trineedle coaxial electrospraying with a simple nonsolvent process. TiO2-Ag nanoparticles and Ganoderma lucidum polysaccharides (GLPs) were encapsulated into the outer shell of the YSPs as the major antibacterial and antioxidant components, whereas iron oxide (Fe3O4) nanoparticles were incorporated into the inner core to act as a photothermal agent. The morphology and structure, chemical composition, biocompatibility, antioxidant, and antibacterial effects of the fabricated YSPs, photothermal effects, and the release profile of the encapsulated GLP were studied in vitro. Furthermore, the in vivo wound healing effects of the YSPs and the laser-assisted therapy were explored based on a burn wound model on c57 mice.

A Case of a Patent Omphalomesenteric Duct Presenting with Meconium Discharge from the Umbilicus

The omphalomesenteric duct is a link between the primitive midgut and the yolk sac. Normally, the duct obliterates around 6 weeks of gestation, yet varying degrees of incomplete obliteration can take place in 1%–4% of infants. This study described the case of a newborn with a patent omphalomesenteric duct remnant fistula identified at birth with meconium in the umbilical cord. At birth, the infant presented meconium staining and meconium discharged within the umbilical cord. Physical examination and other examinations showed no other specific findings. The omphalomesenteric duct fistula was confirmed through the imaging study (abdominal ultrasonography, gastrografin enema). A surgery was carried out where the remnant was resected. The patient did well and was discharged soon after without complication.

Omega-3 polyunsaturated fatty acids promote angiogenesis in placenta derived mesenchymal stromal cells.

Enhancement of angiogenesis is solicited in wound repair and regeneration. Mesenchymal stromal cells derived from the placenta (P-MSCs) have an inherent angiogenic potential. Polyunsaturated fatty acids (PUFAs) in turn, specifically the omega-3 (N-3) are essential for growth and development. They are also recommended as dietary supplements during pregnancy. We therefore hypothesized that addition of N-3 PUFAs in P-MSC culture media may enhance their angiogenic potential. Hence, we treated P-MSCs with omega-3 (N-3) fatty acids -Docosahexaenoic acid (DHA) and Eicosapentaenoic acid (EPA) at different concentrations and tested their angiogenic potential. We saw an upregulation of both bFGF and VEGFA. We also found enhanced in vitro tube formation ability of P-MSCs treated with DHA: EPA. We then looked at the influence of the conditioned medium (CM) collected from P-MSCs exposed to DHA: EPA on the key effector cells -HUVECs (Human Umbilical Vein derived endothelial cells and their functionality was further confirmed on chick yolk sac membrane. We found that the CM of P-MSCs exposed to DHA: EPA could enhance angiogenesis in both cases. These result were finally validated in an in vivo matrigel plug assay which revealed enhanced migration and vessel formation in CM treated with DHA: EPA. Our data thus reveals for the first time that supplementation with lower concentration of PUFA enhances the angiogenic potential of P-MSCs making them suitable for chronic wound healing applications.

Deficient Vitamin E Uptake During Development Impairs Neural Tube Closure in Mice Lacking Lipoprotein Receptor SR-BI.

SR-BI is the main receptor for high density lipoproteins (HDL) and mediates the bidirectional transport of lipids, such as cholesterol and vitamin E, between these particles and cells. During early development, SR-BI is expressed in extraembryonic tissue, specifically in trophoblast giant cells in the parietal yolk sac. We previously showed that approximately 50% of SR-BI-/- embryos fail to close the anterior neural tube and develop exencephaly, a perinatal lethal condition. Here, we evaluated the role of SR-BI in embryonic vitamin E uptake during murine neural tube closure. Our results showed that SR-BI-/- embryos had a very low vitamin E content in comparison to SR-BI+/+ embryos. Whereas SR-BI-/- embryos with closed neural tubes (nSR-BI-/-) had high levels of reactive oxygen species (ROS), intermediate ROS levels between SR-BI+/+ and nSR-BI-/- embryos were detected in SR-BI-/- with NTD (NTD SR-BI-/-). Reduced expression of Pax3, Alx1 and Alx3 genes was found in NTD SR-BI-/- embryos. Maternal α-tocopherol dietary supplementation prevented NTD almost completely (from 54% to 2%, p < 0.001) in SR-BI-/- embryos and normalized ROS and gene expression levels. In sum, our results suggest the involvement of SR-BI in the maternal provision of embryonic vitamin E to the mouse embryo during neural tube closure.

High-precision, non-invasive anti-microvascular approach via concurrent ultrasound and laser irradiation.

Antivascular therapy represents a proven strategy to treat angiogenesis. By applying synchronized ultrasound bursts and nanosecond laser irradiation, we developed a novel, selective, non-invasive, localized antivascular method, termed photo-mediated ultrasound therapy (PUT). PUT takes advantage of the high native optical contrast among biological tissues and can treat microvessels without causing collateral damage to the surrounding tissue. In a chicken yolk sac membrane model, under the same ultrasound parameters (1 MHz at 0.45 MPa and 10 Hz with 10% duty cycle), PUT with 4 mJ/cm2 and 6 mJ/cm2 laser fluence induced 51% (p = 0.001) and 37% (p = 0.018) vessel diameter reductions respectively. With 8 mJ/cm2 laser fluence, PUT would yield vessel disruption (90%, p < 0.01). Selectivity of PUT was demonstrated by utilizing laser wavelengths at 578 nm or 650 nm, where PUT selectively shrank veins or occluded arteries. In a rabbit ear model, PUT induced a 68.5% reduction in blood perfusion after 7 days (p < 0.001) without damaging the surrounding cells. In vitro experiments in human blood suggested that cavitation may play a role in PUT. In conclusion, PUT holds significant promise as a novel non-invasive antivascular method with the capability to precisely target blood vessels.

Epithelial magnesium transport by TRPM6 is essential for prenatal development and adult survival.

Mg2+ regulates many physiological processes and signalling pathways. However, little is known about the mechanisms underlying the organismal balance of Mg2+. Capitalizing on a set of newly generated mouse models, we provide an integrated mechanistic model of the regulation of organismal Mg2+ balance during prenatal development and in adult mice by the ion channel TRPM6. We show that TRPM6 activity in the placenta and yolk sac is essential for embryonic development. In adult mice, TRPM6 is required in the intestine to maintain organismal Mg2+ balance, but is dispensable in the kidney. Trpm6 inactivation in adult mice leads to a shortened lifespan, growth deficit and metabolic alterations indicative of impaired energy balance. Dietary Mg2+ supplementation not only rescues all phenotypes displayed by Trpm6-deficient adult mice, but also may extend the lifespan of wildtype mice. Hence, maintenance of organismal Mg2+ balance by TRPM6 is crucial for prenatal development and survival to adulthood.

Specific polyunsaturated fatty acids modulate lipid delivery and oocyte development in C. elegans revealed by molecular-selective label-free imaging.

Polyunsaturated fatty acids (PUFAs) exhibit critical functions in biological systems and their importance during animal oocyte maturation has been increasingly recognized. However, the detailed mechanism of lipid transportation for oocyte development remains largely unknown. In this study, the transportation of yolk lipoprotein (lipid carrier) and the rate of lipid delivery into oocytes in live C. elegans were examined for the first time by using coherent anti-Stokes Raman scattering (CARS) microscopy. The accumulation of secreted yolk lipoprotein in the pseudocoelom of live C. elegans can be detected by CARS microscopy at both protein (~1665 cm(-1)) and lipid (~2845 cm(-1)) Raman bands. In addition, an image analysis protocol was established to quantitatively measure the levels of secreted yolk lipoprotein aberrantly accumulated in PUFA-deficient fat mutants (fat-1, fat-2, fat-3, fat-4) and PUFA-supplemented fat-2 worms (the PUFA add-back experiments). Our results revealed that the omega-6 PUFAs, not omega-3 PUFAs, play a critical role in modulating lipid/yolk level in the oocytes and regulating reproductive efficiency of C. elegans. This work demonstrates the value of using CARS microscopy as a molecular-selective label-free imaging technique for the study of PUFA regulation and oocyte development in C. elegans.

In Vivo Cardiotoxicity Induced by Sodium Aescinate in Zebrafish Larvae.

Sodium aescinate (SA) is a widely-applied triterpene saponin product derived from horse chestnut seeds, possessing vasoactive and organ-protective activities with oral or injection administration in the clinic. To date, no toxicity or adverse events in SA have been reported, by using routine models (in vivo or in vitro), which are insufficient to predict all aspects of its pharmacological and toxicological actions. In this study, taking advantage of transparent zebrafish larvae (Danio rerio), we evaluated cardiovascular toxicity of SA at doses of 1/10 MNLC, 1/3 MNLC, MNLC and LC10 by yolk sac microinjection. The qualitative and quantitative cardiotoxicity in zebrafish was assessed at 48 h post-SA treatment, using specific phenotypic endpoints: heart rate, heart rhythm, heart malformation, pericardial edema, circulation abnormalities, thrombosis and hemorrhage. The results showed that SA at 1/10 MNLC and above doses could induce obvious cardiac and pericardial malformations, whilst 1/3 MNLC and above doses could induce significant cardiac malfunctions (heart rate and circulation decrease/absence), as compared to untreated or vehicle-treated control groups. Such cardiotoxic manifestations occurred in more than 50% to 100% of all zebrafish treated with SA at MNLC and LC10. Our findings have uncovered the potential cardiotoxicity of SA for the first time, suggesting more attention to the risk of its clinical application. Such a time- and cost-saving zebrafish cardiotoxicity assay is very valid and reliable for rapid prediction of compound toxicity during drug research and development.

New development of the yolk sac theory in diabetic embryopathy: molecular mechanism and link to structural birth defects.

Maternal diabetes mellitus is a significant risk factor for structural birth defects, including congenital heart defects and neural tube defects. With the rising prevalence of type 2 diabetes mellitus and obesity in women of childbearing age, diabetes mellitus-induced birth defects have become an increasingly significant public health problem. Maternal diabetes mellitus in vivo and high glucose in vitro induce yolk sac injuries by damaging the morphologic condition of cells and altering the dynamics of organelles. The yolk sac vascular system is the first system to develop during embryogenesis; therefore, it is the most sensitive to hyperglycemia. The consequences of yolk sac injuries include impairment of nutrient transportation because of vasculopathy. Although the functional relationship between yolk sac vasculopathy and structural birth defects has not yet been established, a recent study reveals that the quality of yolk sac vasculature is related inversely to embryonic malformation rates. Studies in animal models have uncovered key molecular intermediates of diabetic yolk sac vasculopathy, which include hypoxia-inducible factor-1α, apoptosis signal-regulating kinase 1, and its inhibitor thioredoxin-1, c-Jun-N-terminal kinases, nitric oxide, and nitric oxide synthase. Yolk sac vasculopathy is also associated with abnormalities in arachidonic acid and myo-inositol. Dietary supplementation with fatty acids that restore lipid levels in the yolk sac lead to a reduction in diabetes mellitus-induced malformations. Although the role of the human yolk in embryogenesis is less extensive than in rodents, nevertheless, human embryonic vasculogenesis is affected negatively by maternal diabetes mellitus. Mechanistic studies have identified potential therapeutic targets for future intervention against yolk sac vasculopathy, birth defects, and other complications associated with diabetic pregnancies.

Transition of maternal dietary n-3 fatty acids from the yolk to the liver of broiler breeder progeny via the residual yolk sac.

The aim of the present study was to evaluate the transfer of maternal dietary fatty acids (FA) from the yolk to the developing offspring, with special emphasis on n-3 FA eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Six hundred forty Ross 308 breeders were housed from 6 to 58 wk of age in 16 pens resulting in 4 replicates per dietary treatment. They were fed 1 of 4 diets: a basal diet, rich in n-6 FA (CON), or an n-3 FA enriched diet formulated to obtain an EPA/DHA ratio of 1/1 (EPA=DHA), 1/2 (DHA), or 2/1 (EPA). At 28, 43, and 58 wk of age, 20 eggs per treatment were collected and analyzed for FA composition. At these same breeder ages, 600 fertilized eggs per treatment were incubated. At hatch the residual yolks of 25 chicks per treatment were collected and analyzed for FA composition. At every hatch, 180 chicks per treatment were raised under standard conditions and livers were sampled at d 1, 14, 28, and 38 d for FA analysis. Concentrations of EPA in the yolk and residual yolk of eggs laid by EPA-fed breeders were highest, next-to-highest for EPA=DHA-fed breeders, next-to-lowest for DHA-fed breeders, and lowest in those laid by control hens, reflecting the inclusion levels in the maternal diets. Yolk and residual yolk DHA concentrations, however, were not only elevated due to DHA supplementation, compared with the control diet, but also due to EPA supplementation. Offspring hepatic EPA concentrations were elevated until d 28 in all n-3 enriched groups, whereas hepatic DHA concentrations were only affected by EPA=DHA and DHA supplementation at d 1. No differences were found in hepatic DHA concentrations at later offspring ages. Considering the role of EPA and DHA in early development and growth, the maternal supply of these n-3 FA might improve offspring health and performance.

A novel four-step system for screening angiogenesis inhibitors.

Angiogenesis exhibits a significant effect on tumor progression. Inhibiting angiogenesis may provide significant advantages over currently available therapeutics for cancer therapies thus, the development of a system of screening angiogenesis is essential. In the present study, a novel available system of screening angiogenesis inhibitors by four steps was developed. The chorioallantoic membrane (CAM), yolk sac membrane and early chick embryo blood island assay were initially performed to obtain possible antitumor compounds. The MMTV­PyMT transgenic breast cancer mouse model was used for final screening and to confirm potential antitumor effects. Four angiogenesis inhibitors were isolated from 480 compounds, which were obtained from ICCB known bioactives library, by a combination of the CAM, yolk sac membrane and early chick embryo blood island assay. The MMTV­PyMT mouse was treated with one of four agents and it was demonstrated that the tumor volume was significantly inhibited. These results demonstrate that the four­step screening system is feasible.

Maternal-fetal transfer of selenium in the mouse.

Selenoprotein P (Sepp1) is taken up by receptor-mediated endocytosis for its selenium. The other extracellular selenoprotein, glutathione peroxidase-3 (Gpx3), has not been shown to transport selenium. Mice with genetic alterations of Sepp1, the Sepp1 receptors apolipoprotein E receptor-2 (apoER2) and megalin, and Gpx3 were used to investigate maternal-fetal selenium transfer. Immunocytochemistry (ICC) showed receptor-independent uptake of Sepp1 and Gpx3 in the same vesicles of d-13 visceral yolk sac cells, suggesting uptake by pinocytosis. ICC also showed apoER2-mediated uptake of maternal Sepp1 in the d-18 placenta. Thus, two selenoprotein-dependent maternal-fetal selenium transfer mechanisms were identified. Selenium was quantified in d-18 fetuses with the mechanisms disrupted. Maternal Sepp1 deletion, which lowers maternal whole-body selenium, decreased fetal selenium under selenium-adequate conditions but deletion of fetal apoER2 did not. Fetal apoER2 deletion did decrease fetal selenium, by 51%, under selenium-deficient conditions, verifying function of the placental Sepp1-apoER2 mechanism. Maternal Gpx3 deletion decreased fetal selenium, by 13%, but only under selenium-deficient conditions. These findings indicate that the selenoprotein uptake mechanisms ensure selenium transfer to the fetus under selenium-deficient conditions. The failure of their disruptions (apoER2 deletion, Gpx3 deletion) to affect fetal selenium under selenium-adequate conditions indicates the existence of an additional maternal-fetal selenium transfer mechanism.

Visceral endoderm expression of Yin-Yang1 (YY1) is required for VEGFA maintenance and yolk sac development.

Mouse embryos lacking the polycomb group gene member Yin-Yang1 (YY1) die during the peri-implantation stage. To assess the post-gastrulation role of YY1, a conditional knock-out (cKO) strategy was used to delete YY1 from the visceral endoderm of the yolk sac and the definitive endoderm of the embryo. cKO embryos display profound yolk sac defects at 9.5 days post coitum (dpc), including disrupted angiogenesis in mesoderm derivatives and altered epithelial characteristics in the visceral endoderm. Significant changes in both cell death and proliferation were confined to the YY1-expressing yolk sac mesoderm indicating that loss of YY1 in the visceral endoderm causes defects in the adjacent yolk sac mesoderm. Production of Vascular Endothelial Growth Factor A (VEGFA) by the visceral endoderm is essential for normal growth and development of the yolk sac vasculature. Reduced levels of VEGFA are observed in the cKO yolk sac, suggesting a cause for the angiogenesis defects. Ex vivo culture with exogenous VEGF not only rescued angiogenesis and apoptosis in the cKO yolk sac mesoderm, but also restored the epithelial defects observed in the cKO visceral endoderm. Intriguingly, blocking the activity of the mesoderm-localized VEGF receptor, FLK1, recapitulates both the mesoderm and visceral endoderm defects observed in the cKO yolk sac. Taken together, these results demonstrate that YY1 is responsible for maintaining VEGF in the developing visceral endoderm and that a VEGF-responsive paracrine signal, originating in the yolk sac mesoderm, is required to promote normal visceral endoderm development.

Effects of dietary vitamin B1 (thiamine) and magnesium on the survival, growth and histological indicators in lake trout (Salvelinus namaycush) juveniles.

An interaction of two essential nutrients, thiamine and magnesium (Mg) has been documented in in vitro and in vivo studies in mammalian metabolism. However, the role of this association in poikilothermic vertebrates, such as fish, remains elusive. The purpose of this study was first to investigate the effects of dietary thiamine and Mg, and their interaction in lake trout and second to better understand the mechanism leading to early mortality syndrome (EMS), which is caused by a low thiamine level in embryos of many species of salmonids in the wild. Semi-purified diets (SPD) were prepared to accomplish 2 × 2 factorial design that were either devoid of or supplemented with thiamine mononitrate (20 mg/kg diet), magnesium oxide (700 mg/kg diet), or both. Lake trout alevins at the swim-up stage were fed for 10 wk one of the SPD diets or a commercial diet at the same rate (2.0-1.5%) based on recorded biomass. Our results showed that the concentrations of thiamine in the trunk muscle and Mg of whole body were closely associated with the dietary level of two nutrients. The interaction of low dietary Mg and thiamine resulted in apparently worsened overt symptoms of thiamine deficiency in lake trout leading to a higher mortality of fish during the seven week long trial (P<0.05). The fish fed a thiamine-devoid and Mg-supplemented diet were presumed to survive longer (10 wk) than the fish fed diets devoid of both nutrients (discontinued after 7th wk due to high mortality). However, we did not observe histopathological changes in the brain and liver corresponding to thiamine concentrations in tissues. These data suggest that Mg enhanced utilization of the thiamine remaining in the fish body and its interdependence was consistent with observations in mammals. EMS severity might be worsened when Mg is deficient in parental diets (and consequently in yolk sac) and/or first feed.