Project description:Uterine glands are essential for pregnancy in mice and likely humans, because they secrete or transport bioactive substances that regulate uterine receptivity for blastocyst implantation. In mice, the uterus becomes receptive to blastocyst implantation on day 4, but is refractory by day 5. Here, blastocysts could be recovered from progesterone-induced uterine gland (PUGKO) but not wildtype (WT) mice on day 5 post-mating. Anti-adhesive Muc1 protein and microvilli were present on the luminal epithelium of PUGKO but not WT uteri. A number of known uterine receptivity genes and gland-specific genes were altered in the PUGKO uterus. Next, the uterus and uterine luminal fluid (ULF) were obtained from WT and PUGKO mice on day 3, 4 and 5. Transcriptome analysis revealed that 580 genes were decreased in the PUGKO uterus, however ULF secrotome analysis revealed that many proteins and several amino acids were increased in the PUGKO ULF. Of note, many proteins encoded by many gland-specific genes were not identified in the ULF of WT mice. These results support the ideas that uterine glands secrete factors that regulate ULF homeostasis and interact with other cell types in the uterus to influence uterine receptivity and blastocyst implantation for the establishment of pregnancy.
Project description:RNA-seq for the uterine circular muscle (CM) and longitudinal muscle (LM) layers at the mesometrial (M) side on days 12 (pre-implantation stage) and 15 (implantation stage) of pregnancy and day 15 of the estrous cycle in pigs.
Project description:Embryo implantation is a complex process which involves biochemical and physiological interactions between an implantation-competent blastocyst and a receptive uterus. However, the exact biochemical changes of uterine fluid, uterus, and plasma during peri-implantation remain unclear. This study aims to characterize the biochemical and metabolic changes that occur during the peri-implantation period of early pregnancy, using mice as an animal model. Gas chromatography-mass spectrometry was used to analyze the metabolite profiles of the uterus, uterine fluid, and maternal plasma at pre-implantation and implantation. The multivariate analyses, ANOVA and Tukey's HSD test, were applied to detect significant changes in metabolites and metabolic pathways. The metabolic networks were reconstructed in silico based on the identified metabolites and KEGG metabolic framework. Between pre-implantation day 1 and day 4, dramatic metabolic changes were observed in the uterine fluid that could be important for blastocyst development and protection against the harsh uterine environment. Palmitoleic acid, fumaric acid, and glutaric acid changed levels at day 4 in the uterus, suggesting that they may be associated with endometrial receptivity. Both the uterus and maternal plasma showed profound changes in cellular metabolism at the early implantation period, including upregulation of branched-chain amino acids and intermediates of one-carbon metabolism, an upregulation of glyoxylate and dicarboxylate metabolism, and downregulation of aerobic respiration; all of which could be involved in the regulation of the maternal-fetal interface, alternative nutrient utilization, and energy preservation for implantation as well as later placentation and fetal development to ensure successful embryo implantation.
Project description:Dairy cow subfertility is a worldwide issue arising from multiple factors. It manifests in >30% early pregnancy losses in seasonal pasture-grazed herds, especially when cows are inseminated in the early post-partum period. Most losses occur before implantation, when embryo growth depends on factors present in maternal tract fluids. Here we used label-free LC-MS/MS to examine the proteomic composition of uterine luminal fluid in 87 day-7 pregnant cows to identify potential biomarkers of fertility. We also determined changes in uterine luminal fluid from first to third oestrus cycles postpartum in individual cows and linked those changes with divergent embryo development. Out of 1563 proteins detected, 472 had not been previously reported in this fluid, and 408 were predicted to be actively secreted by bioinformatic analysis. The abundance of 19 proteins with roles in immune regulation and metabolic function was associated with contrasting embryo quality (e.g. cystatin B, pyruvate kinase M2). Matched-paired pathway analysis indicated that, from first to third oestrus postpartum, upregulation of metabolic (e.g. creatine and carbohydrate) and immune (e.g. complement system regulation and antiviral defence) processes were related to poorer quality embryos in the third oestrus cycle postpartum. Conversely, upregulated signal transduction and protein trafficking appeared related to improved embryo quality in third oestrus. These results advance the characterisation of the molecular environment of bovine uterine luminal fluid and may aid in understanding fertility issues in other mammals, including humans.
Project description:Progesterone (P4) acts via the endometrium to modify the uterine environment and promotes conceptus growth for elongation and pregnancy establishment. Ewes were ovariectomized and treated with P4 for 14 days or P4 for 14 days and RU486, a progesterone receptor antagonist, from days 8 to 14. Small RNA sequencing of endometrium and EVs from the uterine lumen detected expression of 768 miRNAs and P4 regulation of 9 endometrial and 7 extracellular vesicle miRNAs.
Project description:The protein cargos of uterine extracellular vesicles isolated from uterine lavages, collected from pregnant mares (P; day 10, 11, 12 and 13 after insemination) and cyclic control mares (C; day 10 and 13 after insemination), was analyzed.
Project description:Healthy cow uterine fluid (UF) was collected from synchronized cattle at day 0, 7 and 16 after ovulation. Extracellular vesicles (EVs) were isolated from UF. UF-EVs from same cattle on different timepoints were subjected to LC-MS/MS. Additionally, one sample before EV purification was subjected to LC-MS/MS (corresponding EV sample is named 1D0).
Project description:Progesterone (P4) acts via the endometrium to modify the uterine environment and promotes conceptus growth for elongation and pregnancy establishment. Ewes were ovariectomized and treated with P4 for 14 days or P4 for 14 days and RU486, a progesterone receptor antagonist, from days 8 to 14. Interrogation of the endometrial transcriptome resulted in 1,611 differentially abundant transcripts with 931 increased and 680 decreased in P4 as compared to P4+RU treated ewes.
Project description:Gallus gallus avian eggshell is a biomineral composed of 95% of calcium carbonate on calcitic form and of 3.5% of an extracellular organic matrix (proteins, polysaccharides and proteoglycans). The calcification process occurred in the lumen of the uterus into the acellular uterine fluid, and is divided in three stages (initial, growth and terminal), where the concentration and the nature of proteins varies depending on stages. We have used mass spectrometry methodologies to identify and quantify proteins secreted into the uterine fluid proteins during the three stages of biomineralization.
Project description:The objective of the study was to compare the microRNA content in uterine fluid from patients with recurrent implantation failure (RIF) to that of healthy fertile women. It is a descriptive laboratory study including healthy fertile women and patients with RIF, defined as three failed in vitro fertilization cycles with high quality embryos. Study subjects were instructed to monitor their menstrual cycles using a luteinizing hormone test kit. Uterine fluid was collected on day LH+ 7-9 by flushing with saline. Samples were processed for small RNA sequencing and results were analysed using bioinformatics. The main outcome measure was to identify differentially expressed miRNAs between patients with RIF and healthy fertile women.