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PACS (phosphofurin acidic cluster sorting) proteins are known for their roles in sorting cargo proteins to organelles and can physically interact with WD40 repeat-containing protein WDR37. PACS1, PACS2, and WDR37 variants are associated with multisystemic syndromes and neurodevelopmental disorders characterized by intellectual disability, seizures, developmental delays, craniofacial abnormalities, and autism spectrum disorder. However, the functional effects of syndromic variants at the cellular level remain unknown. Here, we report the expression pattern of Caenorhabditis elegans orthologs of PACS and WDR37 and their interaction. We show that cePACS-1 and ceWDR-37 colocalize to somatic cytoplasm of many types of cells and are mutually required for expression, supporting a conclusion that the intermolecular dependence of PACS1/PACS2/PACS-1 and WDR37/WDR-37 is evolutionarily conserved. We further show that editing in PACS1 and PACS2 variants in cePACS-1 changes protein localization in multiple cell types, including neurons. Moreover, expression of human PACS1 can functionally complement C. elegans PACS-1 in neurons, demonstrating conserved functions of the PACS-WDR37 axis in an invertebrate model system. Our findings reveal effects of human variants and suggest potential strategies to identify regulatory network components that may contribute to understanding molecular underpinnings of PACS/WDR37 syndromes.

From single-cell RNA-sequencing (scRNA-seq) and spatial transcriptomics (ST), one can extract high-dimensional gene expression patterns that can be described by intercellular communication networks or decoupled gene modules. These two descriptions of information flow are often assumed to occur independently. However, intercellular communication drives directed flows of information that are mediated by intracellular gene modules, in turn triggering outflows of other signals. Methodologies to describe such intercellular flows are lacking. We present FlowSig, a method that infers communication-driven intercellular flows from scRNA-seq or ST data using graphical causal modeling and conditional independence. We benchmark FlowSig using newly generated experimental cortical organoid data and synthetic data generated from mathematical modeling. We demonstrate FlowSig's utility by applying it to various studies, showing that FlowSig can capture stimulation-induced changes to paracrine signaling in pancreatic islets, demonstrate shifts in intercellular flows due to increasing COVID-19 severity and reconstruct morphogen-driven activator-inhibitor patterns in mouse embryogenesis.

In single-cell RNA sequencing (scRNA-Seq), gene expression is assessed individually for each cell, allowing the investigation of developmental processes, such as embryogenesis and cellular differentiation and regeneration, at unprecedented resolution. In such dynamic biological systems, cellular states form a continuum, e.g., for the differentiation of stem cells into mature cell types. This process is often represented via a trajectory in a reduced-dimensional representation of the scRNA-Seq dataset. While many methods have been suggested for trajectory inference, it is often unclear how to handle multiple biological groups or conditions, e.g., inferring and comparing the differentiation trajectories of wild-type and knock-out stem cell populations. In this manuscript, we present condiments, a method for the inference and downstream interpretation of cell trajectories across multiple conditions. Our framework allows the interpretation of differences between conditions at the trajectory, cell population, and gene expression levels. We start by integrating datasets from multiple conditions into a single trajectory. By comparing the cells conditions along the trajectorys path, we can detect large-scale changes, indicative of differential progression or fate selection. We also demonstrate how to detect subtler changes by finding genes that exhibit different behaviors between these conditions along a differentiation path.

Non-invasively evaluating gene expression products in human pre-implantation embryos remains a significant challenge. Here, we develop a non-invasive method for comprehensive characterization of the extracellular RNAs (exRNAs) in a single droplet of spent media that was used to culture human in vitro fertilization embryos. We generate the temporal extracellular transcriptome atlas (TETA) of human pre-implantation development. TETA consists of 245 exRNA sequencing datasets for five developmental stages. These data reveal approximately 4,000 exRNAs at each stage. The exRNAs of the developmentally arrested embryos are enriched with the genes involved in negative regulation of the cell cycle, revealing an exRNA signature of developmental arrest. Furthermore, a machine-learning model can approximate the morphology-based rating of embryo quality based on the exRNA levels. These data reveal the widespread presence of coding gene-derived exRNAs at every stage of human pre-implantation development, and these exRNAs provide rich information on the physiology of the embryo.

The Hedgehog (Hh) pathway plays a crucial role in embryonic development, acting both as a morphogenic signal that organizes tissue formation and a potent mitogenic signal driving cell proliferation. Dysregulated Hh signaling leads to various developmental defects in the brain. This article aims to review the roles of Hh signaling in the development of the neocortex in the mammalian brain, focusing on its regulation of neural progenitor proliferation and neuronal production. The review will summarize studies on genetic mouse models that have targeted different components of the Hh pathway, such as the ligand Shh, the receptor Ptch1, the GPCR-like transducer Smo, the intracellular transducer Sufu, and the three Gli transcription factors. As key insights into the Hh signaling transduction mechanism were obtained from mouse models displaying neural tube defects, this review will also cover some studies on Hh signaling in neural tube development. The results from these genetic mouse models suggest an intriguing hypothesis that elevated Hh signaling may play a role in the gyrification of the brain in certain species. Additionally, the distinctive production of GABAergic interneurons in the dorsal cortex in the human brain may also be linked to the extension of Hh signaling from the ventral to the dorsal brain region. Overall, these results suggest key roles of Hh signaling as both a morphogenic and mitogenic signal during the forebrain development and imply the potential involvement of Hh signaling in the evolutionary expansion of the neocortex.

Mammalian ferredoxin 1 and 2 (FDX1/2) belong to an evolutionary conserved family of iron-sulfur cluster containing proteins and act as electron shutters between ferredoxin reductase (FDXR) and numerous proteins involved in critical biological pathways. FDX1 is involved in biogenesis of steroids and bile acids, Vitamin A/D metabolism, and lipoylation of tricarboxylic acid (TCA) cycle enzymes. FDX1 has been extensively characterized biochemically but its role in physiology and lipid metabolism has not been explored. In this study, we generated Fdx1-deficient mice and showed that knockout of both alleles of the Fdx1 gene led to embryonic lethality. We also showed that like Fdxr+/-+/-, Fdx1+/-+/- had a shorter life span and were prone to steatohepatitis. However, unlike Fdxr+/-+/-, Fdx1+/-+/- were not prone to spontaneous tumors. Additionally, we showed that FDX1 deficiency led to lipid droplet accumulation possibly via the ABCA1-SREBP1/2 pathway. Specifically, untargeted lipidomic analysis showed that FDX1 deficiency led to alterations in several classes of lipids, including cholesterol, triacylglycerides, acylcarnitines, ceramides, phospholipids and lysophospholipids. Taken together, our data indicate that FDX1 is essential for mammalian embryonic development and lipid homeostasis at both cellular and organismal levels.

Riboflavin deficiency (RD) induces liver damage, abnormal embryonic development, and high mortality. We hypothesized that the phenotype could be rescued by inhibiting ER stress. The objectives of the present study were to investigate the underlying molecular mechanisms of RD-induced embryonic defects using in vitro and in vivo models. Primary duck embryonic hepatocytes were treated with an ER stress inhibitor (4-PBA) or transfected with CHOP siRNA , and cultured in RD medium and riboflavin-sufficient (CON) medium for 8 days. Laying ducks (n = 20 cages/diet, 1 bird/cage) were fed an RD diet or CON diet for 14 wk, and the eggs were collected for hatching. At day 7 of incubation, the fertilized RD eggs were injected with or without 4-PBA into the yolk. RD decreased cell number and cell viability compared to the CON group, induced oxidative stress and apoptosis in primary duck embryonic hepatocytes. However, after being treated with an ER stress inhibitor (4-PBA) or transfected with CHOP siRNA , the apoptosis rate in RD hepatocytes decreased by 60.6 % and 86.1 %, respectively, being equal to the CON. These results indicated that RD-induced hepatocyte apoptosis is mediated by ER stress and the CHOP pathway. In vivo , RD embryos showed low hatchability, abnormal development, liver damage, ER stress, and apoptosis compared to the CON group. However, 4-PBA administration, as a model of ER stress inhibition, substantially restored embryonic development and alleviated liver damage in the RD group, including ER stress and apoptosis. Notably, hatchability in the RD group increased from 21.7 % to 72.7 % after 4-PBA treatment, though it remained less than the CON group (87.7 %). These results implicated ER stress-CHOP-apoptosis pathway as molecular mechanisms underlying RD-induced abnormal embryonic development and death, this target with potential for therapy or intervention. [Display omitted] • RD leads to liver damage, abnormal embryonic development, and increased mortality in duck embryos. • Riboflavin may modulate ER stress and apoptosis, offering a novel perspective for treating related diseases. • ER stress-CHOP-apoptosis pathways may represent a novel target for regulating embryonic development. [ABSTRACT FROM AUTHOR]

Background: Angiogenesis is indispensable for the sustained survival and progression of both embryonic development and tumorigenesis. This intricate process is tightly regulated by a multitude of pro‐angiogenic genes. The presence of gene modules facilitating angiogenesis has been substantiated in both embryonic development and the context of tumor proliferation. However, it remains unresolved whether the pro‐angiogenic gene modules expressed during embryonic development also exist in tumors. Methods: This study performed a pan‐cancer single‐cell RNA sequencing (scRNA‐seq) analysis on samples from 332 patients across seven cancer types: thyroid carcinoma, lung cancer, breast cancer, hepatocellular carcinoma, colorectal cancer, ovarian carcinoma, and prostate adenocarcinoma. Data processing was carried out using the Seurat R package, with rigorous quality control to filter high‐quality cells and mitigate batch effects across datasets. We used principal component analysis (PCA), shared nearest neighbor graph‐based clustering, and Uniform Manifold Approximation and Projection (UMAP) to visualize cell types and identify distinct cell clusters. Myeloid cell subpopulations were further analyzed for the expression of embryonic pro‐angiogenic gene modules (EPGM) and tumor pro‐angiogenic gene modules (TPGM). Results: The analysis identified nine major cell types within the tumor microenvironment, with myeloid cells consistently exhibiting elevated expression of both tumor pro‐angiogenic gene modules (TPGM) and EPGM across all tumor types. In particular, myeloid cells, including macrophages and monocytes, showed high EPGM expression, indicating an active role of embryonic pro‐angiogenesis pathways in tumors. A subset analysis revealed 20 distinct myeloid subtypes with varying EPGM and TPGM expression across different cancers. Treatment and disease stage influenced these gene expressions, with certain subtypes, such as HSPAhi/STAT1+ macrophages in breast cancer, displaying reduced pro‐angiogenic gene activity post‐treatment. Conclusion: This study provides evidence that tumors may exploit EPGM to enhance vascularization and support sustained growth, as evidenced by the elevated EPGM expression in tumor‐associated myeloid cells. The consistent presence of EPGM in TAMs across multiple cancer types suggests a conserved mechanism wherein tumors harness embryonic angiogenic pathways to facilitate their progression. Distinct EPGM expression patterns in specific myeloid cell subsets indicate potential therapeutic targets, particularly in cases where EPGM activation contributes to resistance against anti‐angiogenic therapies. These findings shed new light on the molecular mechanisms underlying tumor angiogenesis and highlight the prognostic relevance of EPGM expression in cancer, underscoring its potential as a biomarker for clinical applications. [ABSTRACT FROM AUTHOR]

Diapause is a survival strategy for freshwater crustaceans in the genus Daphnia that involves a genetically encoded inhibition of organism growth, development and reproduction. While the environmental triggers for Daphnia to produce diapause‐destined embryos and those that induce hatching are well documented, the influence of environmental challenges during different diapause sub‐phases remains unexplored.In this study, we exposed diapause‐destined embryos of Daphnia magna to different temperatures and light conditions during designated diapause sub‐phases.Our results underscore the importance of the maternal environment for the embryos that are in preparation for diapause. Moreover, we demonstrate a period of obligate developmental arrest during which reinitiation of development cannot be triggered. Additionally, we illustrate the embryos' ability to adjust hatching in response to environmental changes during the hatching process.This study reveals the inherent developmental pattern in Daphnia embryos as they progress through diapause and their adaptability to environmental changes. [ABSTRACT FROM AUTHOR]

The growing variety of RNA classes, such as mRNAs, lncRNAs, and circRNAs, plays pivotal roles in both developmental processes and various pathophysiological conditions. Nonetheless, our comprehension of RNA functions in live organisms remains limited due to the absence of durable and effective strategies for directly influencing RNA levels. In this study, we combined the CRISPR-RfxCas13d system with sperm-like stem cell-mediated semi-cloning techniques, which enabled the suppressed expression of different RNA species. This approach was employed to interfere with the expression of three types of RNA molecules: Sfmbt2 mRNA, Fendrr lncRNA, and circMan1a2(2,3,4,5,6). The results confirmed the critical roles of these RNAs in embryonic development, as their loss led to observable phenotypes, including embryonic lethality, delayed embryonic development, and embryo resorption. In summary, our methodology offers a potent toolkit for silencing specific RNA targets in living organisms without introducing genetic alterations. [ABSTRACT FROM AUTHOR]

Background: Congenital diaphragmatic hernia (CDH) is a rare disease that affects the development of the diaphragm, leading to abnormal lung development. Unfortunately, there is no established therapy for CDH. Retinoic acid pathways are implicated in the ethology of CDH and macrophages are known to play a role in repairing organ damage. Methods: We have analyzed the effect of several Toll like receptor (TLR) ligands in the nitrofen-induced CDH model in pregnant rats widely used to study this disease and in the G2-GATA4Cre;Wt1fl/fl CDH genetic mice model. Morphometric and histological studies were carried out. Immune cell infiltration was assayed by immunochemistry and immunofluorescence and retinoic pathway gene expression analyzed in vivo and in vitro in macrophages. Results: We found that administering a single dose of atypical TLR2/4 ligands (CS1 or CS2), 3 days after nitrofen, cured diaphragmatic hernia in 73% of the fetuses and repaired the lesion with complete diaphragm closure being on the other hand nontoxic for the mothers or pups. Moreover, these immunomodulators also improved pulmonary hypoplasia and alveolar maturation and vessel hypertrophy, enhancing pulmonary maturity of fetuses. We also found that CS1 treatment rescued the CDH phenotype in the G2-GATA4Cre;Wt1fl/fl CDH genetic mice model. Only 1 out of 11 mutant embryos showed CDH after CS1 administration, whereas CDH prevalence was 70% in untreated mutant embryos. Mechanistically, CS1 stimulated the infiltration of repairing M2 macrophages (CD206+ and Arg1+) into the damaged diaphragm and reduced T cell infiltration. Additionally, those TLR ligands induced retinol pathway genes, including RBP1, RALDH2, RARα, and RARβ, in the affected lungs and the diaphragm and in macrophages in vitro. Conclusions: Our research has shown that TLR ligand immunomodulators that influence anti-inflammatory macrophage activation can be effective in treating CDH, being nontoxic for the mothers or pups suggesting that those TLR ligands are a promising solution for CDH leading to orphan drug designation for CS1. The immune system of the fetus would be responsible for repairing the damage and closure of the hernia in the diaphragm and enhanced proper lung development after CS1 treatment. [ABSTRACT FROM AUTHOR]

To protect the germplasm resources of Gymnodiptychus dybowskii (Kessler, 1874), a species endemic to Xinjiang, the key stages of embryonic development of G. dybowskii were studied. From May to July 2021, the artificial breeding of G. dybowskii was carried out at the fish breeding station of the Kenswart Water Conservancy Project. The embryos and early larvae were obtained using temperature promotion, water stimulation, hormone induction, and dry insemination techniques. their development process was observed, and morphological characteristics of each developmental stage were recorded. The results showed that the mature oocyte was round, heavy, and light yellow, with an egg diameter of (2.32 ± 0.21) mm. The fertilized egg was slightly viscous, and the viscosity disappeared after water absorption. Under the temperature of 15.6-17.7 °C (16.23 ± 0.52) °C, the embryonic development of G. dybowskii lasted for 142 h and 33 min and went through 7 stages of fertilization, cleavage, blastula, gastrula, neurula embryo, organ formation, and membrane hatching, with a total of 26 periods. The membrane broke after 2376.724 °C-h. The embryonic development of G. dybowskii follows the rule of freshwater bony fish, but it differs from other Schizothorax species in the muscular effect period and membrane emergence mode. In addition, differences in hatching water temperature among different geographic populations lead to differences in embryo development duration and accumulated temperature, which further proves that G. dybowskii is highly adaptive to the ecological environment of its habitat water, and these characteristics together constitute its survival and reproduction strategy in the natural environment. In this study, through the artificial breeding experiment of G. dybowskii, the characteristics of its embryonic development were explored, which laid the foundation for the cultivation of unique fish seedlings, enriched the early life history data of G. dybowskii, and further provided basic data for its resource protection. [ABSTRACT FROM AUTHOR]

Simple Summary: The in vitro culture of mammalian embryos lacks different embryotrophic factors that are present in the female reproductive tract and are essential for correcting embryonic development. It has been reported that supplementation with oviductal fluid (OF) and uterine fluid (UF) during the in vitro culture of mammalian embryos produced by IVF has a positive effect on their development and quality. We investigated the effect of OF–UF supplementation on the balance of reactive oxygen species (ROS), glutathione (GSH), and the development of cloned and parthenogenetic Ovis aries embryos. However, at concentrations of 1 and 2% OF–UF, the blastocyst rate decreased in both groups of embryos. At a concentration of 1% OF–UF, both intracellular ROS and GSH decreased in the blastocysts of parthenogenetic embryos but not in cloned embryos. In cloned sheep embryos, OF–UF had no impact on the embryonic development or redox balance. Somatic cell nuclear transfer (SCNT) has great potential for the replication of high-commercial-value animals, threatened wild species for conservation purposes, and transgenic animals for biomedical purposes. However, SCNT has a low success rate due to intrinsic factors of the technique itself, which leads to low rates of embryonic development and epigenetic alterations in cloned embryos. The objective of this study was to evaluate the effect of OF–UF on the intracellular concentrations of ROS and GSH and the development of cloned and parthenogenetic Ovis aries embryos. The results do not show a beneficial effect on the development of parthenogenetic and cloned embryos at concentrations of 0.5% OF–UF. Furthermore, at 1% OF–UF, an adverse effect was observed in cloned embryos at the blastocyst stage and 2% OF and UF in parthenogenetic embryos during the first divisions. Decreases in ROS and GSH levels were observed in the parthenogenetic blastocysts treated with 1% OF–UF, but not in the clones, in which a higher concentration of GSH and a similar concentration of ROS were observed. No effect of OF–UF was observed on embryonic development and redox balance in sheep embryos cloned via handmade cloning. [ABSTRACT FROM AUTHOR]

Simple Summary: A comprehensive report about the embryonic development of Cirrhinus mrigala is portrayed. Cirrhinus mrigala embryonic development is classified into 22 stages. A time-lapse imaging technique was utilized for a better understanding and display of the results. The current findings will provide profound insights into future research endeavors and the large-scale seed production of C. mrigala.Cirrhinus mrigala is an important edible fish with a significant aquaculture contribution in Southeast Asian countries. The current study aims to enhance our understanding of the developmental biology of Cirrhinus mrigala, which is crucial for implementing sustainable fish farming practices. To induce spermiation and ovulation in Cirrhinus mrigala brooders, the synthetic hormone Ovaprim® (GnRH + dopamine inhibitor) was administrated as a single injection dose of 0.2 mL/kg to males and 0.4 mL/kg to females. After induction, the fish spawned, and the eggs produced were fertilized artificially and cell division commenced successfully. The characteristics of each larval developmental stage were closely observed and recorded using a time-lapse imaging technique. The fertilized eggs were spherical, demersal, and non-adhesive throughout their incubation period. The spawned eggs ranged in diameter from 2.1 mm to 2.13 mm and possessed circular yolk sacs. The gastrula stage initiated approximately 4 h after fertilization, with 25% of the yolk sphere covered by blastoderm, reaching 75% coverage at the end of the gastrula stage, approximately 6 h post fertilization. Organogenesis was marked by the formation of notochord and the visibility of rudimentary organs such as the heart, eyes, and gills, followed by tail movement, which was observed at the time of hatching. Compared to other cyprinid fishes, C. mrigala exhibited distinct features at certain stages of embryonic development. Blood circulation was observed to start at the onset of hatching. The lengths of the newly hatched larvae ranged from 2.9 to 3.2 mm, smaller than other reports on induced breeding in carps. The findings of the present study provide a detailed reference for the embryonic development of C. mrigala, which will assist its future research endeavors and large-scale seed production for sustainable aquaculture. [ABSTRACT FROM AUTHOR]

The hedgehog signaling pathway plays an important role in early development and growth of most vertebrates. Sonic hedgehog (shh) gene is a critical regulator of embryonic development in many species, including humans. However, it is not clear what roles shh can play in the development of fish. In this paper, shh gene was cloned from Pseudopleuronectes yokohamae. The full‐length complementary DNA (cDNA) of P. yokohamae sonic hedgehog gene (Pyshh) comprises 3194 bp, with a 1317‐bp open reading frame (ORF) that encodes a polypeptide of 438 amino acids with a typical HH‐signal domain and Hint‐N domain. The conserved sequences of the protein among species were predicted by using multiple sequence comparison. The phylogenetic tree construction showed that PySHH is clustered in a branch of Pleuronectidae. To explore the expression of Pyshh gene in various tissues of P. yokohamae, we used real‐time fluorescence quantitative PCR technology to detect it. The results showed that Pyshh gene is widely distributed in various tissues of P. yokohamae juveniles, different tissues of adult males and females, and is particularly expressed in immune organs. The Pyshh gene expression was higher in the muscle and brain of juvenile fish, and higher in bone, gill, and skin of male fish than that of female fish, suggesting that Pyshh might be involved in the formation of immune organs of P. yokohamae. The expression of Pyshh gene significantly upregulated from the gastrula stage to the hatching stage. Western blotting of the expression levels of PySHH during different embryonic development stages revealed that PySHH levels increased gradually during development stages from oosperm stage to hatching stage. These results indicate that Pyshh is highly conserved among species and plays a critical role in the complex process of embryonic development. Its precise regulation is essential for the proper formation of many organs and tissues in the body, and disruptions in its function may have serious consequences for the formation of immune organs in fish. [ABSTRACT FROM AUTHOR]

During embryonic development of the retina of the eye, astrocytes, a type of glial cell, migrate over the retinal surface and form a dynamic mesh. This mesh then serves as scaffolding for blood vessels to form the retinal vasculature network that supplies oxygen and nutrients to the inner portion of the retina. Astrocyte spreading proceeds in a radially symmetric manner over the retinal surface. Additionally, astrocytes mature from astrocyte precursor cells (APCs) to immature perinatal astrocytes (IPAs) during this embryonic stage. We extend a previously-developed continuum model that describes tension-driven migration and oxygen and growth factor influenced proliferation and differentiation. Comparing numerical simulations to experimental data, we identify model equation components that can be removed via model reduction using approximate Bayesian computation (ABC). Our results verify experimental studies indicating that the choroid oxygen supply plays a negligible role in promoting differentiation of APCs into IPAs and in promoting IPA proliferation, and the hyaloid artery oxygen supply and APC apoptosis play negligible roles in astrocyte spreading and differentiation. [ABSTRACT FROM AUTHOR]

Tetraploids play a crucial role in the large-scale production of all-triploid Pacific oyster, Crassostrea gigas by mating with diploids. For nursery farms, research related to larval tolerance to the environment appears to be extremely important. For this goal, fertilization, embryonic development, growth, and settlement from fertilization stage to metamorphosis stage of the diploid, triploid, and tetraploid larvae of C. gigas "Haida No. 2" strain were studied under controlled conditions of temperature (20, 24, 28, and 32 ℃) and salinity (18, 21, 24, 27, and 30 psu). The results showed that increasing temperature and lowering salinity reduced survival time of sperm while increasing temperature and raising salinity accelerated the germinal vesicle breakdown (GVBD) ratio of stripped eggs in diploids and tetraploids for the first time. The appropriate condition for hatching of four crosses (DD, DT, TD, and TT, males were listed first, D for diploid, T for tetraploid) were listed as follows: 20–28 ℃/27–30 psu for DD, 24 ℃/27–30 psu for DT, 24–28 ℃/27–30 psu for TD, and 24 ℃/27–30 psu for TT. Diploids and triploids had similar performance but were all better than tetraploids under different temperature and salinity. The settlement rate increased with rising temperature while salinity and ploidy had little influence on settlement. The information obtained in this study can contribute to increasing the yield of triploid and tetraploid "Haida No. 2" of C. gigas. [ABSTRACT FROM AUTHOR]

Although supplementation with docosahexaenoic acid (DHA) during porcine oocyte IVM is well-established, the available data are limited due to the lack of consistency. Moreover, to our knowledge, the anti-oxidant effects of DHA on porcine oocytes have not been reported. Hence, this study aimed to examine the effects of DHA supplementation on the regulation of energy metabolism during porcine oocyte maturation to improve oocyte maturation and embryonic development. By supplementing the IVM medium with various DHA concentrations, 25 μM DHA was identified as the optimal concentration which improved intraoocyte glutathione content and enhanced embryonic development after parthenogenesis. Compared to embryos derived from the control group, those derived from SCNT or IVF showed significantly improved blastocyst formation upon DHA supplementation during IVM. In addition, various transcription factors associated with oocyte development and apoptosis in mature oocytes were beneficially regulated in the DHA-treated oocytes. Moreover, DHA improved the AMP-activated protein kinase (AMPK)-regulatory ability of porcine oocytes and ameliorated nuclear maturation and embryonic development, which were decreased by artificially downregulating AMPK. To our knowledge, this is the first study to examine the effects of DHA as an AMPK regulator on oocyte maturation and embryo development in pigs. Furthermore, DHA addition to the IVM medium upregulated the relative expression of genes associated with mitochondrial potential and lipid metabolism. Therefore, the membrane potential of mitochondria (evaluated based on the JC-1 aggregate/JC-1 monomer ratio) and the levels of fatty acids and lipid droplets in matured oocytes increased, resulting in increased ATP synthesis. In conclusion, the DHA treatment of porcine oocytes with 25 μM DHA during IVM enhances the homeostasis of energy metabolism by improving mitochondrial function and lipid metabolism, leading to improved quality of matured oocytes and enhanced embryonic developmental potential of in vitro produced (IVP) embryos. Thus, 25 μM DHA supplementation could serve as a tool for improving the quality of IVP embryos. The study findings provide a basis for further research on improving the production efficiency of cloned animals by securing high-quality matured oocytes and enhancing energy metabolism in mammalian oocytes, including those of pigs. • DHA (25 μM) optimally improves porcine oocyte quality and embryonic development. • DHA maintains the homeostasis of energy metabolism of porcine oocytes. • DHA improves pivotal functions of mitochondria in matured porcine oocytes. • DHA supplementation during porcine oocyte maturation improves lipid metabolism. [ABSTRACT FROM AUTHOR]

In view of the rapidly expanding medical and commercial applications of silver nanoparticles (AgNPs), their potential health risks and environmental effects are a significant growing concern. Earlier research by our group uncovered the embryotoxic potential of AgNPs, showing detrimental impacts of these nanoparticles on both pre- and post-implantation embryonic development. In the current study, we showed that low (50–100 μM) and high (200–400 μM) dose ranges of AgNPs trigger distinct cell death programs affecting mouse embryo development and further explored the underlying mechanisms. Treatment with low concentrations of AgNPs (50–100 μM) triggered ROS generation, in turn, inducing mitochondria-dependent apoptosis, and ultimately, harmful effects on embryo implantation, post-implantation development, and fetal development. Notably, high concentrations of AgNPs (200–400 μM) evoked more high-level ROS generation and endoplasmic reticulum (ER) stress-mediated necrosis. Interestingly, pre-incubation with Trolox, a strong antioxidant, reduced ROS generation in the group treated with 200–400 μM AgNPs to the level induced by 50–100 μM AgNPs, resulting in switching of the cell death mode from necrosis to apoptosis and a significant improvement in the impairment of embryonic development. Our findings additionally indicate that activation of PAK2 is a crucial step in AgNP-triggered apoptosis and sequent detrimental effects on embryonic development. Based on the collective results, we propose that the levels of ROS generated by AgNP treatment of embryos serve as a critical regulator of cell death type, leading to differential degrees of damage to embryo implantation, post-implantation development and fetal development through triggering apoptosis, necrosis or other cell death signaling cascades. [ABSTRACT FROM AUTHOR]

After normal pollination and fertilization of pseudoparthenocarpic seedless grapes, their embryos often stop developing due to certain developmental factors, resulting in embryo abortion. Hybrid breeding using seedless grapes as the maternal parent requires embryo rescue breeding technology. This technology plays a crucial role in seedless grape breeding. Although previous studies have extensively explored this technology, knowledge regarding its impact on embryo abortion and the effectiveness of rescue techniques remains limited. This study aimed to investigate the correlation between embryo rescue and hormonal changes in seedless grapes. Four Eurasian seedless grape cultivars, "Thompson Seedless," "Flame Seedless," "Heshi Seedless," and "Ruby Seedless," were selected for examination. We investigated endogenous hormone levels, including indole-3-acetic acid (IAA), gibberellic acid (GA3), and abscisic acid (ABA), in both berries and in vitro ovules during the most suitable embryo rescue time for these cultivars. Based on the observed fluctuations in endogenous hormone levels and previous research findings, appropriate doses of exogenous hormones, such as IAA, GA3, and ABA, were applied during seedless grape embryo rescue. The results indicated significant differences in endogenous hormone levels between berrieswith varying ovule counts of the same cultivar and ovules cultured in vitro, suggesting a hormonal influence on ovule abortion and embryo development in seedless grapes. Further research has identified effective ratios of exogenous hormones: 30 mg·L-1 IAA + 30 mg·L-1 ABA for berry ovule development, 1.0 mg·L-1 IAA + 2.0 mg·L-1 6-benzylaminopurine (6-BA) + 1.0 mg·L-1 GA3 + 1.0 mg·L-1 ABA for in vitro ovule development, and 1.0 mg·L-1 IAA + 2.0 mg·L-1 6-BA + 1.0 mg·L-1 GA3 for embryo germination and seedling formation. In summary, hormonal changes significantly influence ovule and embryo development and are closely related to seedless grape embryo rescue breeding. This study deepened our understanding of the correlation between seedless grape embryo rescue and hormonal changes. It also resulted in the successful production of a batch of embryo rescue seedlings, further improving embryo rescue breeding technology and providing new germplasm materials for developing new seedless grape cultivars. [ABSTRACT FROM AUTHOR]

The rise of "fast fashion" has driven up the production of low-cost, short-lived clothing, significantly increasing global textile fiber production and, consequently, exacerbating environmental pollution. This study investigated the ecotoxicological effects of different types of anthropogenic microfibers—cotton, polyester, and mixed fibers (50% cotton: 50% polyester)—on marine organisms, specifically sea urchin embryos. All tested fibers exhibited toxicity, with cotton fibers causing notable effects on embryonic development even at environmentally relevant concentrations. The research also simulated a scenario where microfibers were immersed in seawater for 30 days to assess changes in toxicity over time. The results showed that the toxicity of microfibers increased with both concentration and exposure duration, with polyester being the most toxic among the fibers tested. Although synthetic fibers have been the primary focus of previous research, this study highlights that natural fibers like cotton, which are often overlooked, can also be toxic due to the presence of harmful additives. These natural fibers, despite decomposing faster than synthetic ones, can persist in aquatic environments for extended periods. The findings underline the critical need for further research on both natural and synthetic microfibers to understand their environmental impact and potential threats to marine ecosystems and sea urchin populations. [ABSTRACT FROM AUTHOR]

SETD2 is the only enzyme responsible for transcription-coupled histone H3 lysine 36 trimethylation (H3K36me3). Mutations in SETD2 cause human diseases including cancer and developmental defects. In mice, Setd2 is essential for embryonic vascular remodeling. Given that many epigenetic modifiers have recently been found to possess noncatalytic functions, it is unknown whether the major function(s) of Setd2 is dependent on its catalytic activity or not. Here, we established a site-specific knockin mouse model harboring a cancer patient-derived catalytically dead Setd2 (Setd2-CD). We found that the essentiality of Setd2 in mouse development is dependent on its methyltransferase activity, as the Setd2CD/CD and Setd2−/− mice showed similar embryonic lethal phenotypes and largely comparable gene expression patterns. However, compared with Setd2−/−, the Setd2CD/CD mice showed less severe defects in allantois development, and single-cell RNA-seq analysis revealed differentially regulated allantois-specific 5′ Hoxa cluster genes in these two models. Collectively, this study clarifies the importance of Setd2 catalytic activity in mouse development and provides a new model for comparative study of previously unrecognized Setd2 functions. [ABSTRACT FROM AUTHOR]

Hippo-YAP/TAZ and Wnt/β-catenin signaling pathways, by controlling proliferation, migration, cell fate, stemness, and apoptosis, are crucial regulators of development and tissue homeostasis. We employed zebrafish embryos as a model system to elucidate in living reporter organisms the crosstalk between the two signaling pathways. Co-expression analysis between the Wnt/β-catenin Tg(7xTCF-Xla.Siam:GFP)ia4 and the Hippo-Yap/Taz Tg(Hsa.CTGF:nlsmCherry)ia49 zebrafish reporter lines revealed shared spatiotemporal expression profiles. These patterns were particularly evident in key developmental regions such as the midbrain–hindbrain boundary (MHB), epidermis, muscles, neural tube, notochord, floorplate, and otic vesicle. To investigate the relationship between the Wnt/β-catenin pathway and Hippo-Yap/Taz signaling in vivo, we conducted a series of experiments employing both pharmacological and genetic strategies. Modulation of the Wnt/β-catenin pathway with IWR-1, XAV939, or BIO resulted in a significant regulation of the Yap/Taz reporter signal, highlighting a clear correlation between β-catenin and Yap/Taz activities. Furthermore, genetic perturbation of the Wnt/β-catenin pathway, by APC inhibition or DKK1 upregulation, elicited evident and robust alteration of Yap/Taz activity. These findings revealed the intricate regulatory mechanisms underlying the crosstalk between the Wnt/β-catenin and Hippo-Yap/Taz signaling, shedding light on their roles in orchestrating developmental processes in vivo. [ABSTRACT FROM AUTHOR]

Synchronization of coupled oscillators is a universal phenomenon encountered across different scales and contexts, e.g., chemical wave patterns, superconductors, and the unison applause we witness in concert halls. The existence of common underlying coupling rules defines universality classes, revealing a fundamental sameness between seemingly distinct systems. Identifying rules of synchronization in any particular setting is hence of paramount relevance. Here, we address the coupling rules within an embryonic oscillator ensemble linked to vertebrate embryo body axis segmentation. In vertebrates, the periodic segmentation of the body axis involves synchronized signaling oscillations in cells within the presomitic mesoderm (PSM), from which somites, the prevertebrae, form. At the molecular level, it is known that intact Notch-signaling and cell-to-cell contact are required for synchronization between PSM cells. However, an understanding of the coupling rules is still lacking. To identify these, we develop an experimental assay that enables direct quantification of synchronization dynamics within mixtures of oscillating cell ensembles, for which the initial input frequency and phase distribution are known. Our results reveal a "winner-takes-it-all" synchronization outcome, i.e., the emerging collective rhythm matches one of the input rhythms. Using a combination of theory and experimental validation, we develop a coupling model, the "Rectified Kuramoto" (ReKu) model, characterized by a phase-dependent, nonreciprocal interaction in the coupling of oscillatory cells. Such nonreciprocal synchronization rules reveal fundamental similarities between embryonic oscillators and a class of collective behaviors seen in neurons and fireflies, where higher-level computations are performed and linked to nonreciprocal synchronization. [ABSTRACT FROM AUTHOR]

Stat3 is a critical transcription factor involved in regulating embryonic development，cell proliferation, apoptosis, inflammation and angiogenesis. The Stat3 protein exists in two isoforms: the fulllength Stat3α and the truncated Stat3β, which lacks the C-terminal transcriptional activation domain. Homozygous mutations in Stat3 in mice lead to early embryonic lethality, which can be rescued by the expression of Stat3β. To further understand the structure and function of the Stat3 protein, we expressed various truncated versions of Stat3 in yeast and demonstrated that Stat3 (aa2-11) could activate transcription in yeast and also function effectively in mammalian cells by performing yeast two-hybrid experiments and dual luciferase assay. Our study offers a potential explanation for how Stat3β expression can reverse the embryonic lethality observed in Stat3 gene mutant mice and enhances our understanding of the structure and function of the Stat3 protein. [ABSTRACT FROM AUTHOR]

Embryo transfer has become a major method to improve fertility in both humans and cattle. The current review focuses on predicting an embryo with a high developmental competence and high potential to establish pregnancy. One way to evaluate the embryo quality is to assess its morphology. However, this approach provides only limited and inadequate information. Using a time-lapse system allows a continuous monitoring of embryonic development. Here we introduce the main morphokinetic parameters and discuss their potential to predict embryo viability, implantation, pregnancy, and live birth. Special attention is given to the association between the transcript's profile and the embryo developmental potential; normally cleaved embryos differ in their profile from their abnormally cleaved counterpart; synchronously and asynchronously cleaved embryos, which are both defined as suitable for transfer, differ in their transcript profile. Recently, the advancements and wide use of time-lapse systems led to the development of algorithms for embryo selection. This technology is already implanted in human in vitro fertilization units, but it is not yet used for domestic animals. The authors believe that combining information from both the human and bovine morphokinetics might reveal the benefits of using a time-lapse system to select embryos with good potential for transfer. [ABSTRACT FROM AUTHOR]

Simple Summary: This study explores how the autumn-spawning bitterling, Acheilognathus rhombeus, reproduces and develops within its mussel host. The research highlights how these fish lay their eggs inside mussels, which then serve as a safe environment for the eggs to develop over winter. A critical phase, called diapause, allows the embryos to pause their development for about seven months, ensuring they survive the cold winter months. Once the temperature rises above 10 °C, development resumes. During this time, tiny structures on the embryos called minute tubercles help anchor them within the mussel, preventing them from being expelled prematurely. These findings provide valuable insights into the complex relationship between the bitterlings and their mussel hosts, revealing important evolutionary adaptations that aid in the reproductive success and survival of these fish. We investigated the reproductive strategies and embryonic development of Acheilognathus rhombeus (a bitterling species that spawns in autumn) within its freshwater mussel host in the Bongseo Stream, South Korea. By focusing on survival mechanisms during critical stages of embryonic development, the selective use of mussel gill demibranchs by the bitterlings and associated adaptive traits were observed over 1 year. A significant diapause phase occurs at developmental stage D, which lasts for approximately 7 months, allowing embryos to survive winter. Development resumes when the temperature exceeds 10 °C. Minute tubercles on the embryos (crucial for anchoring within the host gill demibranchs and preventing premature ejection) exhibit the largest height during diapause, and the height decreases when developmental stage E is reached, when growth resumes. Acheilognathus rhombeus embryos were observed in 30.5% of the mussels, mostly within the inner gills, thereby maximizing spatial use and oxygen access to enhance survival. These results highlight the intricate relationship between A. rhombeus and its mussel hosts, demonstrating the evolutionary adaptations that enhance reproductive success and survival. This study provides valuable insights into the ecological dynamics and conservation requirements of such symbiotic relationships. [ABSTRACT FROM AUTHOR]

Anthropogenically induced changes to the natural world are increasingly exposing organisms to stimuli and stress beyond that to which they are adapted. In aquatic systems, it is thought that certain life stages are more vulnerable than others, with embryos being flagged as highly susceptible to environmental stressors. Interestingly, evidence from across a wide range of taxa suggests that aquatic embryos can hatch prematurely, potentially as an adaptive response to external stressors, despite the potential for individual costs linked with underdeveloped behavioural and/or physiological functions. However, surprisingly little research has investigated the prevalence, causes and consequences of premature hatching, and no compilation of the literature exists. Here, we review what is known about premature hatching in aquatic embryos and discuss how this phenomenon is likely to become exacerbated with anthropogenically induced global change. Specifically, we (1) review the mechanisms of hatching, including triggers for premature hatching in experimental and natural systems; (2) discuss the potential implications of premature hatching at different levels of biological organisation from individuals to ecosystems; and (3) outline knowledge gaps and future research directions for understanding the drivers and consequences of premature hatching. We found evidence that aquatic embryos can hatch prematurely in response to a broad range of abiotic (i.e. temperature, oxygen, toxicants, light, pH, salinity) and biotic (i.e. predators, pathogens) stressors. We also provide empirical evidence that premature hatching appears to be a common response to rapid thermal ramping across fish species. We argue that premature hatching represents a fascinating yet untapped area of study, and the phenomenon may provide some additional resilience to aquatic communities in the face of ongoing global change. [ABSTRACT FROM AUTHOR]

Problem: Predicting the impact of systemic inflammation on oocyte and embryonic development in unexplained infertile women using the new immunological indexes. Method of Study: This retrospective cohort study was conducted using the records of the In Vitro Fertilization Department of Ankara Gülhane Training and Research Hospital. After reviewing the records of patients who had undergone in vitro fertilization (IVF) for unexplained infertility (UI) and excluding all known factors that could cause systemic immune inflammation, the systemic immune response index (SIRI), and pan‐immune score were calculated from the pre‐treatment hemogram parameters between the embryo arrest (EA) group and the embryo transfer group. It was investigated whether there was a statistical difference between the two groups and whether an SIRI value affecting embryo quality was found. A receiver operating characteristic (ROC) curve analysis was performed to determine the optimal cut‐off values for inflammatory markers to predict EA. Results: The 108 EA group (embryos that were arrested during their development and could not be transferred) and the 140 embryo transfer group showed statistically significant differences in the parameters of systemic inflammatory index (SII), SIRI, pan‐immune inflammation value (PIV), and neutrophil/lymphocyte ratio (NLR) (p < 0.05). These inflammatory parameters, which were examined before ovulation induction, also correlated positively with the required total dose of gonadotropin and negatively with the ovarian sensitivity index (OSI). SII, SIRI, PIV, and NLR have specific cut‐off values with ROC analysis and determine the effect of the inflammatory status of the environment in which the oocyte develops on EA (p < 0.005). Conclusion: In women with UI, high levels of systemic immune inflammation have a negative impact on oocyte and embryo development, and treatments to suppress inflammation may improve IVF success. [ABSTRACT FROM AUTHOR]

Purpose of review: The cardiac conduction system, composed of pacemaker cells and conducting cardiomyocytes, orchestrates the propagation of electrical activity to synchronize heartbeats. The conduction system plays a crucial role in the development of cardiac arrhythmias. In the embryo, the cells of the conduction system derive from the same cardiac progenitors as the contractile cardiomyocytes and and the key question is how this choice is made during development. Recent Findings: This review focuses on recent advances in developmental biology using the mouse as animal model to better understand the cellular origin and molecular regulations that control morphogenesis of the cardiac conduction system, including the latest findings in single-cell transcriptomics. Summary: The conducting cell fate is acquired during development starting with pacemaking activity and last with the formation of a complex fast-conducting network. Cardiac conduction system morphogenesis is controlled by complex transcriptional and gene regulatory networks that differ in the components of the cardiac conduction system. [ABSTRACT FROM AUTHOR]

Objective: Taking advantage of the broad coverage of Wireless Application Protocol (WAP), we developed a Content Management System (CMS)-programmed mobile learning application. This application can help the undergraduate to obtain a comprehensive understanding of concepts in Cleft lip and palate Phenotype, and Embryonic development (CPE). The present study aims to evaluate the feasibility and efficacy of the concept acquisition teaching model on the basis of WAP in a practical undergraduate course of CPE. Design: The concept acquisition teaching model based on WAP was programmed by CMS, covering definitions of various cleft lip and palate, the mechanisms underlying the phenotypes, practical medical cases, as well as corresponding tests after learning. Setting: The CPE concept acquisition teaching model was distributed to a total of 524 undergraduate students and 46 tutors participated in CPE teaching from seven highly ranked schools of stomatology in China since April 2022. Participants: 524 undergraduate students and 46 tutors from seven highly ranked schools of stomatology in China. Interventions: The CPE concept acquisition teaching model. Main outcome measures: The effectiveness of the CPE teaching model. Results: The response rate to the survey was 100%. The grading of the questionnaires indicated that the students were satisfied with the usability, practicality, and outcome, whereas the tutors were more positive with the contents, cooperation, and outcome. Conclusions: The present study demonstrated the feasibility and efficacy of the WAP-based concept acquisition teaching model of CPE and a high level of satisfaction among undergraduate students and tutors who major in Stomatology. [ABSTRACT FROM AUTHOR]

Abstract Background Congenital diaphragmatic hernia (CDH) is a rare disease that affects the development of the diaphragm, leading to abnormal lung development. Unfortunately, there is no established therapy for CDH. Retinoic acid pathways are implicated in the ethology of CDH and macrophages are known to play a role in repairing organ damage. Methods We have analyzed the effect of several Toll like receptor (TLR) ligands in the nitrofen-induced CDH model in pregnant rats widely used to study this disease and in the G2-GATA4Cre;Wt1fl/fl CDH genetic mice model. Morphometric and histological studies were carried out. Immune cell infiltration was assayed by immunochemistry and immunofluorescence and retinoic pathway gene expression analyzed in vivo and in vitro in macrophages. Results We found that administering a single dose of atypical TLR2/4 ligands (CS1 or CS2), 3 days after nitrofen, cured diaphragmatic hernia in 73% of the fetuses and repaired the lesion with complete diaphragm closure being on the other hand nontoxic for the mothers or pups. Moreover, these immunomodulators also improved pulmonary hypoplasia and alveolar maturation and vessel hypertrophy, enhancing pulmonary maturity of fetuses. We also found that CS1 treatment rescued the CDH phenotype in the G2-GATA4Cre;Wt1fl/fl CDH genetic mice model. Only 1 out of 11 mutant embryos showed CDH after CS1 administration, whereas CDH prevalence was 70% in untreated mutant embryos. Mechanistically, CS1 stimulated the infiltration of repairing M2 macrophages (CD206+ and Arg1+) into the damaged diaphragm and reduced T cell infiltration. Additionally, those TLR ligands induced retinol pathway genes, including RBP1, RALDH2, RARα, and RARβ, in the affected lungs and the diaphragm and in macrophages in vitro. Conclusions Our research has shown that TLR ligand immunomodulators that influence anti-inflammatory macrophage activation can be effective in treating CDH, being nontoxic for the mothers or pups suggesting that those TLR ligands are a promising solution for CDH leading to orphan drug designation for CS1. The immune system of the fetus would be responsible for repairing the damage and closure of the hernia in the diaphragm and enhanced proper lung development after CS1 treatment.

The TGF-β regulatory system plays crucial roles in the preservation of organismal integrity. TGF-β signaling controls metazoan embryo development, tissue homeostasis, and injury repair through coordinated effects on cell proliferation, phenotypic plasticity, migration, metabolic adaptation, and immune surveillance of multiple cell types in shared ecosystems. Defects of TGF-β signaling, particularly in epithelial cells, tissue fibroblasts, and immune cells, disrupt immune tolerance, promote inflammation, underlie the pathogenesis of fibrosis and cancer, and contribute to the resistance of these diseases to treatment. Here, we review how TGF-β coordinates multicellular response programs in health and disease and how this knowledge can be leveraged to develop treatments for diseases of the TGF-β system.

There is emerging evidence that the cardiac interatrial septum has an important role as a thromboembolic source for ischemic strokes. There is little consensus on treatment of patients with different cardiac interatrial morphologies or pathologies who have had stroke. In this paper, we summarize the important background, diagnostic, and treatment considerations for this patient population as presented during the Federation of American Societies for Experimental Biology (FASEB) Virtual Catalytic Conference on the Cardiac Interatrial Septum and Stroke Risk, held on December 7, 2022. During this conference, many aspects of the cardiac interatrial septum were discussed. Among these were the embryogenesis of the interatrial septum and development of anatomic variants such as patent foramen ovale and left atrial septal pouch. Also addressed were various mechanisms of injury such as shunting physiologies and the consequences that can result from anatomic variants, as well as imaging considerations in echocardiography, computed tomography, and magnetic resonance imaging. Treatment options including anticoagulation and closure were addressed, as well as an in-depth discussion on whether the left atrial septal pouch is a stroke risk factor. These issues were discussed and debated by multiple experts from neurology, cardiology, and radiology.

Eggshell thickness can be an indicator of environmental pollution in wild birds and shell quality in wild and domestic birds, but it is difficult to measure calcite eggshell thickness due to the presence of the adherent outer eggshell membrane. Eggshells of 13 waterbird species were divided in half longitudinally and the outer membrane was removed from one of the halves. Subsequently, we measured eggshell thickness, both with and without the outer eggshell membrane, using a Hall-effect thickness gauge to the nearest 0.001 mm along the equator of each eggshell half. Outer eggshell membrane thicknesses ranged from 0.014 to 0.073 mm. Caspian Tern (Hydroprogne caspia) and California Gull (Larus californiens) had the thickest eggshell membranes (0.056 and 0.073 mm, respectively), and Green Heron (Butorides virescens) and Killdeer (Charadrius vociferus) had the thinnest membranes (0.014 and 0.022 mm, respectively). The eggshell membrane, as a percent of the total eggshell and membrane thickness, varied among the 13 species and ranged among species from 7.9% to 20.6%. The outer membrane comprised a greater percent of the total eggshell and membrane thickness for Black Skimmer (19.3%; Rynchops niger), California Gull (20.5%), and Forster's Tern (20.6%; Sterna forsteri) than for Green Heron (7.9%), Double-crested Cormorant (10.4%; Phalacrocorax auritus), and Western Grebe (10.6%; Aechmophorus occidentalis). Within species, the outer membrane thickness was not correlated with egg morphometries but, for a subset of species, there was some indication that the calcite eggshell thickness decreases with embryo development (age). We discuss several reasons for conducting future eggshell thickness measurements without removing the membrane. Received 27 February 2023. Accepted 31 December 2023. Key words: eggshell measurement, eggshell membrane, eggshell thickness, Hall-effect thickness gauge. El grosor de la cascara de huevo puede ser indicador de la contaminacion ambiental en aves silvestres y de calidad de la cascara tanto en aves silvestres como domesticas, pero es dificil medir el grosor de la calcita de la cascara de huevo por la presencia de la membrana externa adherente. Cascaras de huevos de 13 especies de aves acuaticas fueron divididas en mitades longitudinales y la membrana externa fue removida de una de las dos mitades. Luego medimos el grosor de las cascaras de huevo, tanto con como sin membrana externa, usando calibrador de grosor de efecto Hall con al 0.001 mm mas cercano, proximo al ecuador de cada mitad de la cascara de huevo. El grosor de la membrana externa vario entre 0.014 to 0.073 mm. El charran Hydroprogne caspia y la gaviota Larus californicus tuvieron las membranas mas gruesas (0.056 and 0.073 mm, respectivamente), mientras que la garcita verde (Butorides virescens) y el chorlo Charadrius vociferus tuvieron las membranas mas delgadas (0.014 y 0.022 mm, respectivamente). La membrana del cascaron como porcentaje de la suma del grosor del cascaron y de la membrana, vario entre las 13 especies de 7.9% a 20.6%. La membrana externa represento un mayor porcentaje del grosor total de cascara de huevo y membrana para rayador Rynchops niger (19.3%), la gaviota (20.5%) y el charran Sterna forsteri (0.6%) que para la garcita verde (7.9%), cormoran Phalacrocorax auritus (10.4%) y archichilique Aechmophorus occidentalis (10.6%). Dentro de las especies, el grosor de la membrana externa no estaba correlacionado con la morfometria del huevo pero, para un subconjunto de especies, hubo algunos indicios de que el grosor de la calcita de la cascara de huevo disminuyo con el desarrollo del embrion (edad). Discutimos varias razones para llevar a cabo mediciones futuras de grosor de cascara de huevo sin remover la membrana. Palabras clave: medida de cascara de huevo, membrana de cascara de huevo, grosor de cascara de huevo, calibrador de grosor de efecto Hall., Eggshell thickness is used as an indicator of environmental pollution in wild birds, eggshell quality in wild and domestic birds (e.g., Blus 2011, Maurer et al. 2012), and to investigate [...]

New technologies have resulted in a better understanding of blood and lymphatic vascular heterogeneity at the cellular and molecular levels. However, we still need to learn more about the heterogeneity of the cardiovascular and lymphatic systems among different species at the anatomical and functional levels. Even the deceptively simple question of the functions of fish lymphatic vessels has yet to be conclusively answered. The most common interpretation assumes a similar dual setup of the vasculature in zebrafish and mammals: a cardiovascular circulatory system, and a lymphatic vascular system (LVS), in which the unidirectional flow is derived from surplus interstitial fluid and returned into the cardiovascular system. A competing interpretation questions the identity of the lymphatic vessels in fish as at least some of them receive their flow from arteries via specialised anastomoses, neither requiring an interstitial source for the lymphatic flow nor stipulating unidirectionality. In this alternative view, the 'fish lymphatics' are a specialised subcompartment of the cardiovascular system, called the secondary vascular system (SVS). Many of the contradictions found in the literature appear to stem from the fact that the SVS develops in part or completely from an embryonic LVS by transdifferentiation. Future research needs to establish the extent of embryonic transdifferentiation of lymphatics into SVS blood vessels. Similarly, more insight is needed into the molecular regulation of vascular development in fish. Most fish possess more than the five vascular endothelial growth factor (VEGF) genes and three VEGF receptor genes that we know from mice or humans, and the relative tolerance of fish to whole‐genome and gene duplications could underlie the evolutionary diversification of the vasculature. This review discusses the key elements of the fish lymphatics versus the SVS and attempts to draw a picture coherent with the existing data, including phylogenetic knowledge. [ABSTRACT FROM AUTHOR]

During embryonic development, vitamins are involved in energy metabolism and protein synthesis, while cathepsins and antioxidant defense play key roles in yolk degradation and protecting fish embryos from oxidative damage. This study aimed to investigate the consumption of vitamins (A, B1, B2, C, D, and E), ontogenesis of cathepsins, and antioxidant capacity in developing embryos of turbot. In this study, embryos were collected over time from 2.3 to 102.5 h (i.e., hatching) after fertilization for vitamins, cathepsins, and antioxidant capacity analyses. Results showed that the embryonic growth rate in the cleavage stage (CS) was faster than in other stages, while that in the segmentation stage (SS) was lower than in other stages. Embryos displayed higher demands on VA during blastula stage (BS), gastrula stage (GS), the early SS, and hatching day (HD), VB1 at CS; VB2 at BS, GS, SS, and HD; VD at CS and SS; VE at CS, BS, and the late SS; and Vc during entire periods. Cathepsins A (CA) showed a high level at early SS, suggesting its involvement in yolk degradation. Cathepsins B (CB) and cathepsins D (CD) were highly active at GS, suggesting an active apoptosis process. CA, CB, CD, and cathepsins L (CL) all kept low levels in CS, revealing low protein demand during the cleavage process. On the whole, the predominant antioxidant enzymes were superoxide dismutase (SOD) and malondialdehyde (MDA) from CS to SS; SOD, MDA, glutathione peroxidase (GSH-Px), and catalase (CAT) during SS; and GSH-Px and CAT during HD. In conclusion, the fastest growth rate and the slowest growth rate of embryos occurred in CS and SS, respectively, being consistent with energy allocation. Vitamin consumption and cathepsin ontogenesis were stage-specific. Oxidative damage on embryos was relatively less at CS, but larger at SS and HD. [ABSTRACT FROM AUTHOR]

Growing evidence suggests that metabolic regulation directly influences cellular function and development and thus may be more dynamic than previously expected. In vivo and in real-time analysis of metabolite activities during development is crucial to test this idea directly. In this study, we employ two metabolic biosensors to track the dynamics of pyruvate and oxidative phosphorylation (Oxphos) during the early embryogenesis of the sea urchin. A pyruvate sensor, PyronicSF, shows the signal enrichment on the mitotic apparatus, which is consistent with the localization patterns of the corresponding enzyme, pyruvate kinase (PKM). The addition of pyruvate increases the PyronicSF signal, while PKM knockdown decreases its signal, responding to the pyruvate level in the cell. Similarly, a ratio-metric sensor, Grx-roGFP, that reads the redox potential of the cell responds to DTT and H 2 O 2 , the known reducer and inducer of Oxphos. These observations suggest that these metabolic biosensors faithfully reflect the metabolic status in the cell during embryogenesis. The time-lapse imaging of these biosensors suggests that pyruvate and Oxphos levels change both spatially and temporarily during embryonic development. Pyruvate level is increased first in micromeres compared to other blastomeres at the 16-cell stage and remains high in ectoderm while decreasing in endomesoderm during gastrulation. In contrast, the Oxphos signal first decreases in micromeres at the 16-cell stage, while it increases in the endomesoderm during gastrulation, showing the opposite trend of the pyruvate signal. These results suggest that metabolic regulation is indeed both temporally and spatially dynamic during embryogenesis, and these biosensors are a valuable tool to monitor metabolic activities in real-time in developing embryos. • PKM and pyruvate localize on the spindle and in the nucleus during early embryogenesis of the sea urchin. • Pyruvate is increased in micromeres, while Oxidative phosphorylation is decreased in micromeres. • Pyruvate and Oxidative phosphorylation sensors show an opposite trend of expression dynamics during embryogenesis. • Metabolic biosensors are valuable tools for analyzing in vivo metabolic regulation in developing embryos. [ABSTRACT FROM AUTHOR]

Introduction: The specialists refer unexplained infertility to the so-called diagnosis of exclusion due to the fact that in the process of medical examination of married couples the causes of infertility cannot be established. Material and methods: The clinical characteristics and embryological features of in vitro fertilization programs of couples with unexplained infertility versus the patients with tuboperitoneal infertility were analyzed retrospectively and prospectively. The study group comprised 93 women, who underwent 108 in vitro fertilization programs, and the control group consisted of 45 patients, who underwent 49 in vitro fertilization programs. Results: Significant differences (p<0.05) were found between the groups in anamnestic, clinical, laboratory, and instrumental characteristics. The ovarian stimulation protocols were comparable between the groups of patients. The blastulation rate was considered to be the endpoint in in vitro fertilization programs, and it was significantly lower in the group of women with unexplained infertility (45.53%). In-depth analysis of the embryonic stage of in vitro fertilization programs showed, that low blastulation rate in unexplained infertility was mainly due to the fact that embryos stopped developing about three days after they were cultured. At the same time the morphological assessment showed that the quality of blastocysts was higher in the group of unexplained infertility (66.7%) compared to the group of tuboperitoneal factor of infertility (45.8%). Preimplantation genetic testing for aneuploidy showed similar frequency of detection of euploid embryos (41.7% and 40.0%, respectively). Conclusion: A "clinical portrait" of women with unexplained infertility was described. The low blastulation rate was noted in in vitro fertilization programs for women with unexplained infertility. Given the identified impairments of early embryonic development in unexplained infertility, it is appropriate to recommend the patients to undergo early use of in vitro fertilization with good-quality embryo transfer (>3, AA, AB, BA according to Gardner grading system) on day 5-6 of culture without long-lasting preliminary examination and empirical treatment. [ABSTRACT FROM AUTHOR]

Abstract Background Extensive research has been conducted on embryonic developmental disorders linked to Polycystic Ovary Syndrome (PCOS), a pathological condition that affects 5−10% of women and is characterized by irregularities in the menstrual cycle and infertility. By employing RNA sequencing (RNA-seq), we performed an in-depth investigation of PCOS-related changes in gene expression patterns at the mouse blastocyst stage. Methods The zygotes of female B6D2 mice were obtained and then differentiated into blastocysts in K + Simplex Optimised Medium (KSOM) cultures containing exo-NC (negative control for exosomes) or exo-LIPE-AS1 (a novel exosomal marker of PCOS). Subsequently, blastocysts were collected for RNA-seq. The bioinformatics was performed to analyze and compare the differences of gene expression profile between blastocysts of control and PCOS group. Results There were 1150 differentially expressed genes (DEGs) between the two groups of mouse blastocysts; 243 genes were upregulated and 907 downregulated in the blastocysts of the exo-LIPE-AS1 group compared to those of the exo-NC group. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that the genes involved in amino acid synthesis and glutathione metabolic pathways were down-regulated in exo-LIPE-AS1 group. Conclusion This study has revealed that blastocyst developmental retardation may be associated with the downregulation of amino acid synthesis and glutathione metabolism, which may affect energy metabolism, biosynthesis, cellular osmotic pressure, antioxidant synthesis, ROS clearance or mitochondrial function, and ultimately cause blastocyst cell development abnormalities. Our research offers encouraging data on the mechanisms underlying aberrant embryonic development in patients with PCOS as well as potential treatment strategies.

Embryo transfer has become a major method to improve fertility in both humans and cattle. The current review focuses on predicting an embryo with a high developmental competence and high potential to establish pregnancy. One way to evaluate the embryo quality is to assess its morphology. However, this approach provides only limited and inadequate information. Using a time-lapse system allows a continuous monitoring of embryonic development. Here we introduce the main morphokinetic parameters and discuss their potential to predict embryo viability, implantation, pregnancy, and live birth. Special attention is given to the association between the transcript’s profile and the embryo developmental potential; normally cleaved embryos differ in their profile from their abnormally cleaved counterpart; synchronously and asynchronously cleaved embryos, which are both defined as suitable for transfer, differ in their transcript profile. Recently, the advancements and wide use of time-lapse systems led to the development of algorithms for embryo selection. This technology is already implanted in human in vitro fertilization units, but it is not yet used for domestic animals. The authors believe that combining information from both the human and bovine morphokinetics might reveal the benefits of using a time-lapse system to select embryos with good potential for transfer.

Abstract The ribosomal DNA (rDNA) constitutes a remarkably conserved DNA sequence within species, located in the area of the nucleolus, and responsible for coding three major types of rRNAs (18S, 5.8S and 28S). While historical investigations into rDNA focused on its structure and coding capabilities, recent research has turned to explore its functional roles in various biological processes. In this review, we summarize the main findings of rDNA methylation with embryonic development, aging and diseases in multiple species, including epigenetic alterations, related biological processes and potential applications of rDNA methylation. We present an overview of current related research and identify gaps in this field.

Abstract As the most numerous group of animals on Earth, insects are found in almost every ecosystem. Their useful role in the environment is priceless; however, for humans, their presence may be considered negative or even harmful. For years, people have been trying to control the number of pests by using synthetic insecticides, which eventually causes an increased level of resistance to applied compounds. The effects of synthetic insecticides have encouraged researchers to search for alternatives and thus develop safe compounds with high specificity. Using knowledge about the physiology of insects and the functionality of compounds of insect origin, a new class of bioinsecticides called peptidomimetics, which are appropriately modified insect analogues, was created. One promising compound that might be successfully modified is the thirteen amino acid peptide alloferon (HGVSGHGQHGVHG), which is obtained from the hemolymph of the blue blowfly Calliphora vicinia. Our research aimed to understand the physiological properties of alloferon and the activity of its peptidomimetics, which will provide the possibility of using alloferon or its analogues in the pharmaceutical industry, as a drug or adjuvant, or in agriculture as a bioinsecticide. We used alloferon and its three peptidomimetics, which are conjugates of the native peptide with three unsaturated fatty acids with various chain lengths: caprylic, myristic, and palmitic. We tested their effects on the morphology and activity of the reproductive system and the embryogenesis of the Tenebrio molitor beetle. We found that the tested compounds influenced the growth and maturation of ovaries and the expression level of the vitellogenin gene. The tested compounds also influenced the process of egg laying, embryogenesis, and offspring hatching, showing that alloferon might be a good peptide for the synthesis of effective bioinsecticides or biopharmaceuticals.

Objective·To investigate the effects of the cancer-testis antigen 14 (CT14) on embryonic and larval development in nematodes by using the model organism Caenorhabditis elegans (C. elegans), aiming to uncover its potential functions and mechanisms during development.Methods·Transgenic C. elegans strains were constructed by using microinjection for the inducible expression of human CT14 (HsCT14), a truncated mutant of CT14 (HsCT14∆CIR) lacking CT14-specific intermediate region (CIR), and a green fluorescent protein (GFP) control. The impacts of full-length and truncated mutant CT14 on nematode embryonic and larval development were analyzed and compared. Additionally, transgenic C. elegans strains with inducible expression of CT14 from various primates, including the crab-eating macaque (Macaca fascicularis) and mouse lemur (Microcebus murinus), were also constructed to assess the effects on egg hatching and larval-to-adult transformation rates. The differential gene expression in nematode embryos induced by CT14 was analyzed by Smart-seq transcriptome sequencing, with further insights gained through KEGG (Kyoto Encyclopedia of Genes and Genomes) and GSEA (Gene Set Enrichment Analysis), to explore the involved biological processes and pathways.Results·The induced expression of HsCT14 and its truncated mutant HsCT14∆CIR significantly reduced the hatching rate of nematode eggs, with a more pronounced effect observed in HsCT14-expressing strains. Differential interference contrast (DIC) microscopy imaging revealed significant morphological abnormalities in embryos expressing HsCT14 during the comma stage. Nematodes expressing HsCT14 or HsCT14∆CIR exhibited developmental arrest in larvae and substantially lower larval-to-adult transformation rates compared to the GFP control. The impact was more pronounced in nematodes expressing HsCT14 than those with HsCT14∆CIR. The expression of Macaca fascicularis CT14 (MfCT14) exhibited significant effects on the hatching rate and adult transformation rate, similar to that of HsCT14, while the expression of Microcebus murinus CT14 (MmCT14) displayed significantly reduced impact compared to HsCT14 and MfCT14. Smart-seq results indicated that CT14 expression affected various biological processes in nematode embryos, related to ATP-dependent chromatin remodeling and DNA replication.Conclusion·Ectopic expression of the cancer-testis antigen CT14 significantly disrupts both embryonic and larval developments in C. elegans, with the CIR sequence substantially enhancing this effect. It suggests that CT14 may play an important regulatory role in biological development by affecting gene expression in multiple pathways, including chromatin remodeling.

This study aimed to determine the effect of in ovo injections of garlic and tomato extracts on the embryogenesis and neurological traits of chicken embryos with a focus on growth parameters and neurological development during the embryonic stage. A total of 300 eggs were placed in five distinct groups: Group 1 (control negative, CO, without injection); Group 2 (control positive, distilled water, DW, 0.1 mL/egg); Group 3 (garlic extract, GAR, 0.1 mg/egg); Group 4 (tomato extract, TOM, 0.1 mg/egg); and Group 5 (garlic and tomato extract mix, ATM, 0.1 mL/egg). The narrow ends of the albumins were injected with the extracts on the fifth day of incubation using a 1 mL syringe with a 2-gauge needle. The results show a significant increase (p

Abstract Compound eyes formation in decapod crustaceans occurs after the nauplius stage. However, the key genes and regulatory mechanisms of compound eye development during crustacean embryonic development have not yet been clarified. In this study, RNA-seq was used to investigate the gene expression profiles of Neocaridina denticulata sinensis from nauplius to zoea stage. Based on RNA-seq data analysis, the phototransduction and insect hormone biosynthesis pathways were enriched, and molting-related neuropeptides were highly expressed. There was strong cell proliferation in the embryo prior to compound eye development. The formation of the visual system and the hormonal regulation of hatching were the dominant biological events during compound eye development. The functional analysis of DEGs across all four developmental stages showed that cuticle formation, muscle growth and the establishment of immune system occurred from nauplius to zoea stage. Key genes related to eye development were discovered, including those involved in the determination and differentiation of the eye field, eye-color formation, and visual signal transduction. In conclusion, the results increase the understanding of the molecular mechanism of eye formation in crustacean embryonic stage.

Engineered nanomaterials (ENMs) are ubiquitous in contemporary applications, yet their environmental and human health impacts remain inadequately understood. This study addresses the challenge of identifying potential risks associated with ENM exposure by highlighting the significant variability in existing research methodologies. Without a systematic collection of toxicological data that encompasses standardized materials, relevant platforms, and assays, the task of identifying potential risks linked to ENM exposure becomes an intricate challenge. In vitro assessments often use media rich in ionic species, such as RPMI and fetal bovine serum (FBS). Zebrafish embryos, known to develop normally in low-ionic environments, were exposed to Cerium Oxide, Zinc Oxide, and Graphene Oxides in different media at varying concentrations. Here, we discovered that zebrafish embryos tolerated a mix of 80 % RPMI, 2 % FBS, and 1 % antibiotic cocktail. The results revealed that adverse effects observed in zebrafish with certain nanomaterials in Ultra-Pure (UP) water were mitigated in cell culture medium, emphasizing the importance of revisiting previously considered non-toxic materials in vitro. The zebrafish results underscore the importance of utilizing a multidimensional in vivo platform to gauge the biological activity of nanomaterials accurately.

Waprin, a WAP (Whey acidic protein) domain‐containing extracellular secretory protein, is widely known for its antibacterial properties. In this study, a waprin homologue (Tc_wapF) expressing in a female‐specific manner was identified in Tribolium castaneum, through the analysis of sex‐specific transcriptomes. Developmental‐ and tissue‐specific profiling revealed the widespread expression of Tc_wapF in adult female tissues, particularly in the ovary, gut and fatbody. This female‐specific expression of Tc_wapF is not regulated by the classical sex‐determination cascade of T. castaneum, as we fail to get any attenuation in Tc_wapF transcript levels in Tcdsx and Tctra (key players of sex determination cascade of T. castaneum) knockdown females. RNA interference‐mediated knockdown of Tc_wapF in females led to the non‐hatching of eggs laid by these females, suggesting the crucial role of Tc_wapF in the embryonic development in T. castaneum. This is the first report on the identification of a sex‐specific waprin homologue in an insect and its involvement in embryonic development. Future investigations on the functional conservation of insect waprins and their mechanistic role in embryonic development can be exploited for improving pest management strategies. [ABSTRACT FROM AUTHOR]