The Identification and Functional Analysis of CircRNAs in Endometrial Receptivity of Mice with Polycystic Ovary Syndrome
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ABSTRACT: The disorders of endometrial receptivity and ovulatory dysfunction are both important causes of infertility of patients with Polycystic Ovary Syndrome (PCOS). The study aimed to investigate the expression profile and functional analysis of circRNAs in the endometrial receptivity of mice with PCOS. Twenty-four 4-week-old female C57BL/6 mice were randomly divided into two groups (PCOS group and Normal control group, n=12) and PCOS group was subcutaneously with DHEA 6mg/kg for 20 days. The circRNA expression profile in the endometrium tissue of two groups on the 4th day of gestation was screened by gene chip technique.
Project description:Polycystic ovary syndrome (PCOS) is one of the most common endocrinopathies among fertile women. PCOS patients have been confirmed to be accompanied by an increased risk of pregnancy complication, and the exact cause is still unclear. Many factors may cause this situation, and impaired endometrial receptivity could be responsible for adverse pregnancy outcomes in PCOS. Unfortunately, there were still few researches about the molecular mechanisms regarding impaired endometrial receptivity. So we designed this study to explore the secretory endometrial proteome in PCOS patients.
Project description:Impaired endometrial receptivity is one of the major causes of recurrent implantation failure (RIF), and the underlying molecular mechanism has not been fully elucidated. We demonstrated that Chromodomain Y like (CDYL) was highly expressed in the endometrium at mid-secretory phase during the normal menstrual cycles. However, the expression of CDYL was down-regulated in the endometrial tissues obtained from women with RIF, consistently with the protein level of LIF, which is the marker of endometrial receptivity. Knockdown of CDYL significantly decreased the cell migration of human endometrial Ishikawa cells and primary endometrial cells. Genomic analyses revealed that CDYL gene silencing resulted in decreased expression of many genes associated with cytoskeleton and migration regulation.
Project description:Intrauterine infusion of CXCL12 has a beneficial effect on endometrial angiogenesis by recontruction of endometrial vasculature and improvement of endometrial receptivity
Project description:We find that the Endometrial Blood Vessel Generation is an important part ofthe establishment of Endometrial Receptivity. We have demonstrated inprevious experiments: Acupuncture in IVF-ET treatment can effectively improve Endometrial Receptivity and improve the pregnancy rate, therefore, we proposea VEGF signaling pathway in Acupuncture can improve Endometrial receptivity, thus improving the success rate of IVF-ET Hypothesis. Around the hypothesisintends to adopt the rat model of IVF-ET, through methods of acupuncture, acupuncture combined with blocker, research the related indexes effect performance that acupuncture on the VEGF signaling pathway in endometrial. Using experimental techniques such as Morphological observation, ELIAS, Immunohistochemistry, Real-time quantitative PCR, Methylation-sensitive high-resolution melting curve method(MS-HRM), to complete the research ofobserving the Effect of acupuncture interference IVF-ET, Blocking the impaction VEGF to the effect, the relationship VEGF or gene methylation inits receptors with effect. From the molecular level clarify the adverse effects and mechanism action that acupuncture will improve IVF-ET superovulation of endometrial receptivity, through adjust VEGF signaling pathway, to provide a scientific basis for acupuncture in IVF-ET treatment
Project description:Several lines of evidence indicate that there exists differential display of biochemical characteristics in endometrium during receptivity in the proven conception cycle as compared to corresponding secretory phases endometrium of non-conception cycles of fertile rhesus monkeys. In the present study, we examined the differential display in the whole genome expression of endometrium during days 2, 4 and 6 post-ovuation in proven conception cycles and normo-ovulatory, non-conception cycles in proven fertile rhesus monkeys. Female monkeys showing at least two consecutive ovulatory menstrual cycles of normal length were allocated to either of the two groups: group 1 in which females were allowed to cohabit with proven fertile males during days 8 to 16 of their menstrual cycles, and group 2 in which females were not allowed to cohabit. Vaginal smears were checked daily and the days of ovulation were assessed from immunopositive profiles of estradiol-17β and progesterone in peripheral serum samples. Endometrial samples were collected on days 2, 4 and 6 after ovulation from both groups of animals following laparotomy and fundal hysterectomy under ketamine anaesthesia. In group 1, only endometrial samples obtained from mated females who yielded viable, preimplantation-stage matched embyos on uterine flushing were used. In group 2, endometrial samples were obtained from females who showed normal ovulatory cycles. Tissue samples from both groups were washed in ice-cold PBS and were used for RNA extraction to run rhesus monkey whole genome expression array. On average, ~60% and ~68% genes of total number of hybridized genes were expressed in endometrial samples of groups 1 and 2, respectively. Group-wise hierarchical cluster analysis (HCA) yielded highest segregation between peri-implantation stage (groups 1b and 1c) and other (groups 2a-c and group 1a) samples with cluster distance (cd) 0.9, when cd=1.0 denotes complete segregation. On the other hand, least segregation was seen between groups 1a and 2a with cd 0.2, when cd=0 denotes complete aggregation. The number of common genes were less with the progress in days in group 1 (that is in proven conception cycle) unlike that in group 2 (that is in non-mated, normo-ovulatory group). Also, the number of common genes between groups for a given day after ovulation displayed a clear declining trend with increasing number of uncommon genes. Differential dipslay of regulated common genes among all the subgroups based on statistical analyses of variances in data sets obtained from whole genome expression arrays identified a total of fifteen (15) genes that displayed differential (>2-fold, p< 0.05) expression: ADCY5, ADIPOR1, AVEN, CHRND, FOXD3, GJD4, MAPK8IP3, MKS1, NNMT, NUP50, PALT1, PIGV, TGFBR2, TOX2, VWA5B1.
Project description:To identify a role for protein palmitoylation in ovarian of DHEA-induced PCOS mice model. First, we used DHEA to induce a PCOS mice model,hematoxylin-eosin (H&E) staining sections form the ovaries of the Control and DHEA groups.The mice oral gavage with DHEA disrupted ovarian morphology,the estrous cycle, and hormone profilein DHEA-induced mouse. The three pairs of ovary for each tube are mixed together as a group were lysed and divided into 2 fractions. The -HAM condition served as a negative control and the +HAM sample positive control.In the +HAM sample, the palmitate residue was cleaved off and exchanged with biotin. After the ABE reaction was completed, streptavidin beads were used to enrich for biotinylated proteins. Proteins enriched from +HAM and −HAM conditions were identified using mass spectrometry (MS).
Project description:Embryo implantation is a key step in establishing pregnancy and a major limiting factor in IVF. Implantation requires the endometrium, the inner lining of the uterus, to transform from a non-receptive to a receptive state, to allow embryos to attach to the surface and enter into the tissue. However, the fundamental mechanisms governing receptivity are not well understood. Here we show that transmembrane protein podocalyxin is a major and clinically significant factor regulating human endometrial receptivity. Podocalyxin is expressed in all endometrial epithelial cells in the non-receptive state but is selectively down-regulated in the luminal epithelium at receptivity. We present evidence that podocalyxin critically governs the barrier function of the endometrial epithelium, likely as an intrinsic protective mechanism, rendering it non-receptive to an embryo. In addition, podocalyxin suppresses genes promoting receptivity (eg LIF, CSF1) but stimulates those inhibiting implantation (eg WNT7A, LEFTY2). Down-regulation of podocalyxin in the luminal epithelium, likely mediated by progesterone, selectively converts the endometrial surface to a more adhesive state that facilitates embryo attachment and penetration. Furthermore, inadequate down-regulation of podocalyxin in the endometrial luminal epithelium is associated with poorer implantation rates in IVF. We thus propose that podocalyxin promotes the barrier function of human endometrial epithelial cells and critically regulates receptivity for embryo implantation.
Project description:Different progesterone levels and timing of exposure results in different endometrial gene expression and implantation potential. Sufficient progesterone level is important to induce endometrial receptivity and endometrial expressed the most receptive stage after exposure to progesterone for 7 days based on decidualisation marker expression.