Project description:Embryo DNA fingerprinting represents an important tool for tracking embryo-specific outcomes after multiple embryo transfer during IVF. The situation in which 2 embryos are transferred and only one implants represents a unique opportunity for the most well-controlled comparison of competent and incompetent embryos. Specifically, this design eliminates all patient-related variables from the comparison of embryos with or without reproductive potential. However, in order to determine which embryo implanted, the investigator must wait until newborn DNA is available upon delivery. This study validates a non-invasive fetal DNA fingerprinting method that reduces the time to identify which embryo implanted by approximately 31 weeks. Thirty-four patients were studied to determine if fingerprinting of fetal DNA extracted from maternal plasma at 9 gestational weeks concurred with the buccal DNA results obtained from the newborn after delivery. This validation required single nucleotide polymorphism (SNP) profiles on each couples’ preimplantation embryos, enriched fetal DNA from maternal plasma at 9 weeks gestation, and newborn DNA obtained from buccal swabs after delivery. The predictions from fetal DNA-based embryo tracking and gender assignments made at 9 weeks gestation were 100% consistent with standardized methods of assessment performed after term delivery. This study demonstrates the first validated fetal DNA fingerprinting method which predicts both gender and which embryo implanted at 9 weeks gestation following multiple embryo transfer.
Project description:Embryo DNA fingerprinting represents an important tool for tracking embryo-specific outcomes after multiple embryo transfer during IVF. The situation in which 2 embryos are transferred and only one implants represents a unique opportunity for the most well-controlled comparison of competent and incompetent embryos. Specifically, this design eliminates all patient-related variables from the comparison of embryos with or without reproductive potential. However, in order to determine which embryo implanted, the investigator must wait until newborn DNA is available upon delivery. This study validates a non-invasive fetal DNA fingerprinting method that reduces the time to identify which embryo implanted by approximately 31 weeks. Thirty-four patients were studied to determine if fingerprinting of fetal DNA extracted from maternal plasma at 9 gestational weeks concurred with the buccal DNA results obtained from the newborn after delivery. This validation required single nucleotide polymorphism (SNP) profiles on each couples’ preimplantation embryos, enriched fetal DNA from maternal plasma at 9 weeks gestation, and newborn DNA obtained from buccal swabs after delivery. The predictions from fetal DNA-based embryo tracking and gender assignments made at 9 weeks gestation were 100% consistent with standardized methods of assessment performed after term delivery. This study demonstrates the first validated fetal DNA fingerprinting method which predicts both gender and which embryo implanted at 9 weeks gestation following multiple embryo transfer. Affymetrix SNP arrays were processed and successfully completed according to the manufacturer's directions on DNA extracted from 136 embryos, 33 parental blood samples, 17 enriched fetal DNA samples and 21 buccal DNA samples.
Project description:Triclosan (TCS), an antibacterial compound commonly added to personal care products, could be an endocrine disruptor at low doses. Although TCS has been shown to alter fetal physiology, its effects in the developing fetal brain are unknown. The objective of this study was to use transcriptomics and systems analysis to determine significantly altered biological processes in the late gestation ovine fetal hypothalamus after direct or indirect exposure to low doses of TCS. We found that short-term infusion of TCS induces vigorous changes in the fetal hypothalamic transcriptomics, which are mainly related to food intake pathways and metabolism. For direct TCS exposure, chronically catheterized late gestation fetal sheep were infused with vehicle (n=4) or TCS (250 μg/day; n=4) iv. For indirect TCS exposure, TCS (100 μg/kg/day; n=3) or vehicle (n=3) was infused into the maternal circulation. Fetal hypothalami were collected after 2 days of infusion, and gene expression was measured using Agilent 15k ovine microarrays.
Project description:During the initial maternal recognition of pregnancy (MRP), the equine embryo displays a series of unique events characterized by rapid blastocyst expansion, secretion of a diverse array of molecules, and transuterine migration to interact with the luminal epithelium surface. Up to date, the intricate transcriptome and proteome changes of the embryo underlying these events have not been critically studied in horses. Thus, the objective of this study was to perform an integrative transcriptomic and proteomic analyses of embryos collected from days 10 to 13 of gestation.
Project description:Patients who are diagnosed with thrombotic thrombocytopenic purpura (TTP) during pregnancy are at increased risk of maternal and fetal complications including fetal demise. We present a case of a 32-year-old G3P0 (gravida 3, para 0) who presented at 20 weeks’ gestation with a new diagnosis of congenital TTP (cTTP) and fetal demise. Methods: We describe the pathophysiology of pregnancy complications in a patient with cTTP using platelet procoagulant membrane dynamics analysis and quantitative proteomic studies, compared to 4 pregnant patients with gestational hypertension, 4 pregnant patients with preeclampsia and 4 healthy pregnant controls. Results: The cTTP patient had increased P-selectin, tissue factor expression, annexin-V binding on platelets and neutrophils, and localized thrombin generation, suggestive of hypercoagulability. Among 15 proteins that were upregulated, S100A8 and S100A9 were distinctly overexpressed.Conclusions: There is platelet-neutrophil activation and interaction, platelet hypercoagulability and proinflammation in our case of cTTP with fetal demise.
Project description:The success of early pregnancy depends largely on the successful implantation of the embryo. Dynamic crosstalk between embryo and mother plays a crucial role in this process, but our understanding is still limited. Here, using the pig as a model, we first comprehensively profile the single-cell transcriptome of early free embryos and corresponding maternal endometrium, identifying embryonic and matched endometrial cells with heterogeneity. We developed a novel tool, ExtraCellTalk, to predict crosstalk between the early embryo and the maternal uterus using uterine luminal proteins as bridges, generating an overall dynamic map of maternal-fetal crosstalk. Through cross-species comparisons, we identified a conserved RBP4/STRA6 pathway in which embryonic-derived RBP4 targets the STRA6 receptor on stromal cells in the endometrium, thereby interacting with endothelial and perivascular cells to regulate their interconversion to promote endometrial remodeling. These maternal-fetal crosstalks and corresponding endometrial changes that we uncovered are essential for subsequent embryo implantation and are the basis for further studies to improve embryo implantation.
Project description:We hypothesized that poor maternal nutrition during gestation would reduce the growth and development of offspring muscle prenatally, reduce the number of myogenic progenitor cells, and result in changes in the global expression of genes involved in prenatal muscle development and function. Ewes were fed a control (100% NRC)-, restricted (60% NRC),- or over-fed (140% NRC) diet beginning at day 30 of gestation until day 45, 90, and 135 of gestation or until parturition. In offspring, we report altered secondary: primary muscle fiber ratios and percent PAX7 positive cells during fetal development. In addition, changes in muscle mRNA expression were observed between maternal dietary treatments and over timepoints during gestation. In conclusion, poor maternal nutrition during gestation contributes to altered offspring muscle growth during early fetal development which persists throughout the fetal stage.
Project description:Reduced representation bisulfite sequencing of BVDV PI and controls fetal spleens, collected on day 245 of gestation (maternal inoculation at day 75 of gestation)
Project description:Pregnancy is a time of extreme metabolic demand that requires coordinated adaptations between mother and fetus. To determine the contributions of maternal and fetal metabolism to metabolic plasticity during gestation, mice with a liver-specific Carnitine Palmitoyltransferase-2 knockout mice (Cpt2-/-), or Pparα KO mice were subjected to late-gestation nutrient stress, a 24hr fast from E16.5 to E17.5. The fetal response to maternal fasting was dominated by maternal lipid metabolism as the loss of maternal hepatic fatty acid oxidation or Pparα signaling accelerated fetal liver transcriptional programing. These data show that maternal nutritional environment is a major driver of perinatal metabolic programing and plasticity.
Project description:Early life exposures are critical in fetal programming and may influence function and health in later life. Adequate maternal folate consumption during pregnancy is essential for healthy fetal development and long-term offspring health. The mechanisms underlying fetal programming are poorly understood, but are likely to involve gene regulation. Epigenetic marks, including DNA methylation, regulate gene expression and are modifiable by folate supply. We observed before transcriptional changes in fetal liver in response to maternal folate depletion and hypothesised that these changes are due to altered gene promoter methylation. Female C57BL/6J mice were fed diets containing 2âmg or 0.4âmg folic acid/kg for 4âweeks before mating and throughout pregnancy. At 17.5 day gestation, genome-wide gene expression and promoter methylation were measured by microarray analysis in male fetal livers.