Olfactomedin1 (Olfm1) in fallopian tube may modulate tubal ectopic pregnancy in humans: evidence from Immunohistochemistry and an in vitro coculture model

Conference Theme: The Intersection Between Genetics, Genomics, and Reproductive BiologyOlfactomedins are secretary glycoprotein constituted in the extracellular matrix (ECM) of various cell types. Recent studies suggested that Olfm-1 is down-regulated during the window of implantation (WOI) in the human endometrium and up-regulated in pathological condition like endometriosis and recurrent spontaneous abortions. Ectopic pregnancy is a gynaecological emergency and fertility threatening phenomenon which occurs in 1-2% of normal pregnancies and shows an increasing trend. Yet, tubal ectopic pregnancy accounts for more than 98% of the cases ...postprintThe 43rd Annual Meeting of the Society for the Study of Preproduction (SSR), Milwaukee, WI., 30 July-3 August 2010. In Biology of Reproduction, 2010, v. 83 n. Meeting abstracts, p. 27-28, abstract no. 13

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) suppresses spheroids attachment on endometrial epithelial cells through the down-regulation of the Wnt-signaling pathway

The environmental toxicant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) affects embryo development, implantation and fertility in humans. The underlying molecular mechanism by which TCDD suppresses implantation remains largely unknown. We used the trophoblastic spheroids (embryo surrogate)-endometrial cells co-culture assay to study the attachment of trophoblastic spheroids (BeWo and Jeg-3) onto the endometrial epithelial (RL95-2 and Ishikawa) cells. TCDD dose-dependently induced cytochrome P450 1A1 (Cyp1A1) expression in trophoblastic and endometrial epithelial cells. Moreover, TCDD at 1 and 10. nM suppressed β-catenin (a Wnt-signaling molecule) and E-cadherin expression, as well as spheroids attachment onto endometrial cells. Interestingly, activation of the canonical Wnt-signaling pathway via Wnt3a or lithium chloride reverted the suppressive effect of TCDD on β-catenin and E-cadherin expressions in the BeWo and RL95-2 cells, and restored the spheroids attachment rate to be comparable to the untreated controls. Taken together, TCDD induces Cyp1A1 expression, modulates the Wnt-signaling pathway and suppresses spheroids attachment onto endometrial cells. © 2011 Elsevier Inc.postprin

Hormonal regulation of endometrial olfactomedin expression and its suppressive effect on spheroid attachment onto endometrial epithelial cells

Background Olfactomedin (Olfm) is a member of a diverse group of extracellular matrix proteins important for neuronal growth. Recent microarray studies identified Olfm as one of the down-regulated transcripts in receptive endometrium at the time of embryo attachment and implantation. However, the underlying molecular mechanisms that govern Olfm expression and its effect on embryo attachment and implantation remain unknown. Methods The expression of Olfm in the human endometrium was investigated by real-time PCR, western blotting and immunohistochemistry on human endometrial biopsies from natural and ovarian stimulated cycles. To investigate the function of Olfm in trophoblastendometrial cell attachment, an in vitro spheroid-endometrial cell co-culture study was performed. Results Human endometrial Olfactomedin-1 and -2(Olfm-1 and -2) transcripts decreased significantly from the proliferative to the secretory phases of the menstrual cycle. Olfm protein was strongly expressed in the luminal and glandular epithelium and moderately in the stromal cells of human endometria. Ovarian stimulation significantly decreased (P < 0.05) the expression of endometrial Olfm-1 and -2 transcripts in patients receiving IVF treatment when compared with those in the natural cycle. Importantly, recombinant Olfm-1 suppressed JAr spheroid attachment onto Ishikawa cells and this was not associated with changes of β-catenin and E-cadherin expression in trophoblast and endometrial cells. Conclusions Decreased expression of Olfm during the receptive phase of the endometrium may allow successful trophoblast attachment for implantation. © 2010 The Author.postprin

The endocrine disruptor TCDD modulates microRNA expression in preimplantation mouse embryos and spheroids attachment on human endometrial epithelial cells in vitro

Conference Theme: The Intersection Between Genetics, Genomics, and Reproductive BiologyThe endocrine disruptor (ED) is an exogenous substance that acts on the endocrine system and modulates normal physiological functions of the body. Although EDs such as 2,3,7,8-Tetrachlorodibenzodioxin (TCDD) affect normal reproductive function in humans and affects the growth and reproductive functions in rodents, the underlying mechanism that modulates these changes remains unclear. Accumulating evidence suggested preimplantation embryo development is controlled by ...postprintThe 43rd Annual Meeting of the Society for the Study of Reproduction (SSR), Milwaukee, WI., 30 July-3 August 2010. In Biology of Reproduction, 2010, v. 83 Meeting abstracts, p. 62, abstract no. 27

Excessive ovarian stimulation up-regulates the Wnt-signaling molecule DKK1 in human endometrium and may affect implantation: An in vitro co-culture study

Background: High serum estradiol (E2) levels following ovarian stimulation lead to reduced implantation and pregnancy rates, yet the underlying mechanisms remain unknown. We investigated if aberrant expression of genes in the Wnt-signaling pathway may be involved. Methods: Microarray and real-time PCR analysis were performed to analyze gene expression profiles of endometrial samples taken at day hCG + 7 in stimulated cycles, and days LH + 7 and LH + 10 in natural cycles. Expression of several Wnt-signaling transcripts, including Dickkopf homolog 1 (DKK1), DKK2 and secreted frizzled-related protein 4 (sFRP4), was analyzed throughout the menstrual cycle. JAr spheroid/Ishikawa endometrial cell co-culture experiments were established to study effects of DKK1 on spheroid attachment in vitro. Results: We identified 351 differentially expressed genes. Endometrial samples taken at hCG + 7 had similar expression profiles to those at LH + 10. DKK1 transcripts were up-regulated and DKK2 and sFRP4 were down-regulated in the stimulated compared with LH + 7 group (all P < 0.05). DKK1 transcripts were low in proliferative phase (PS) and increased in late-secretory phase (LS, P < 0.05), although DKK2 peaked in mid-secretory phase (P < 0.05). sFRP4 transcripts were high in PS. Treatment of spheroid with recombinant human DKK-1 protein dose-dependently suppressed (P < 0.05 versus control) spheroids attachment onto endometrial cells (associated with decreased-catenin protein): this suppression was nullified by anti-DKK1 antibody.CONCLUSIONGene expression patterns in stimulated cycles resembled those of LS in natural cycles, when the implantation window is about to close, suggesting high serum E2 and/or progesterone concentrations may advance endometrial development, altering the implantation window and possibly decreasing pregnancy rate. Aberrant expression of DKK1 might impair embryo attachment and implantation in vivo.postprin

Effects of fungicide 'Mancozeb' on bovine spermatozoa

Session - Health and Hygiene: no. P163Fungicides are used to maximize the production and productivity of modern agriculture. Present work was undertaken to investigate the effect/s of Mancozeb fungicide on bovine spermatozoa. Semen samples were obtained from Jersey bulls (n=6) at the Central Artificial Insemination Station, Kundasale. Preliminary studies were carried out to determine the effective dose and incubation time using 0.01μg/ml, 0.1μg/ml, 1μg/ml and 10μg/ml of pure Mancozeb and 1.0 μg/ml commercial Mancozeb concentrations during 1-4 hours of incubation periods. Sperm motility parameters were investigated using a Computer Assisted Sperm Analysis (CASA) system. Further functional analyses were performed to test the Acrosine ...postprin

Artificial Neural Network Inference (ANNI): A Study on Gene-Gene Interaction for Biomarkers in Childhood Sarcomas

Objective: To model the potential interaction between previously identified biomarkers in children sarcomas using artificial neural network inference (ANNI). Method: To concisely demonstrate the biological interactions between correlated genes in an interaction network map, only 2 types of sarcomas in the children small round blue cell tumors (SRBCTs) dataset are discussed in this paper. A backpropagation neural network was used to model the potential interaction between genes. The prediction weights and signal directions were used to model the strengths of the interaction signals and the direction of the interaction link between genes. The ANN model was validated using Monte Carlo cross-validation to minimize the risk of over-fitting and to optimize generalization ability of the model. Results: Strong connection links on certain genes (TNNT1 and FNDC5 in rhabdomyosarcoma (RMS); FCGRT and OLFM1 in Ewing’s sarcoma (EWS)) suggested their potency as central hubs in the interconnection of genes with different functionalities. The results showed that the RMS patients in this dataset are likely to be congenital and at low risk of cardiomyopathy development. The EWS patients are likely to be complicated by EWS-FLI fusion and deficiency in various signaling pathways, including Wnt, Fas/Rho and intracellular oxygen. Conclusions: The ANN network inference approach and the examination of identified genes in the published literature within the context of the disease highlights the substantial influence of certain genes in sarcomas

Olfactomedin 1 (Olfm1): a novel protein modulates the embryo like trophoblastic spheroids attachment to the endometrial cells in vitro

Poster presentations: Theme 6 - Reproduction and Development, Musculoskeletal System: no. P.6.03The 14th Research Postgraduate Symposium, The University of Hong Kong, Hong Kong, 2-3 December 2009