155. UNDERSTANDING THE CROSSTALK IN THE HUMAN UTERINE CAVITY: ROLES FOR SOLUBLE MEDIATORS DURING EMBRYO IMPLANTATION

2010 ◽  
Vol 22 (9) ◽  
pp. 73
Author(s):  
N. Hannan ◽  
P. Paiva ◽  
K. L. Meehan ◽  
C. Hincks ◽  
L. J. F. Rombauts ◽  
...  
Keyword(s):  
Menstrual Cycle ◽  
Embryo Implantation ◽  
Uterine Cavity ◽  
Uterine Receptivity ◽  
Adhesive Properties ◽  
Infertile Women ◽  
Functional Studies ◽  
Glandular Secretion ◽  
Uterine Fluid ◽  
Insight Into

Embryo implantation requires synchronized dialogue between a receptive endometrium and an activated blastocyst via locally produced soluble mediators. During the mid-secretory (MS) phase of the menstrual cycle there is increased glandular secretion into the uterine lumen. These secretions contain important mediators that modulate the endometrium and support the conceptus during implantation. Analysis of the composition of uterine fluid across the menstrual cycle and in fertile and infertile women will, therefore, provide new insights into uterine receptivity. We hypothesized that multiplex platform analysis of human uterine lavages would identify soluble mediators important for the establishment of pregnancy in humans. Lavages were collected (by flushing the uterine cavity with 5mL of saline) from fertile and infertile women during the MS phase and from fertile women during the mid-proliferative (MP) phase of the menstrual cycle. Comparison of lavages from the three cohorts was performed using quantitative MilliplexTM Luminex® cytokine/chemokine assays containing 42-analytes. Luminex analysis detected a number of cytokines in uterine fluid, revealing 8 soluble mediators previously unknown in the endometrium and present in human uterine fluid including, PDGF-AA, TNFβ, sIL-2Rα, Flt-3 ligand, sCD40L, IL-7, IFNα2 and GRO. Furthermore comparison of the three cohorts revealed VEGF levels were significantly higher in the fertile (MS) fluid when compared to infertile. Functional studies demonstrated that rhVEGF treatment significantly increased the adhesive properties in cells present at the maternal-fetal interface. These findings suggest VEGF plays a role in regulating embryo implantation. Furthermore identifying the soluble mediators in uterine fluid may provide potential markers of endometrial receptivity, insight into the unique microenvironment essential for pregnancy and a profile of maternal factors that influence the implanting blastocyst.

532. SOLUBLE MEDIATORS IN THE HUMAN UTERINE CAVITY: POTENTIAL ROLES IN EMBRYO IMPLANTATION

2009 ◽  
Vol 21 (9) ◽  
pp. 130
Author(s):  
N. Hannan ◽  
K. L. Meehan ◽  
L. J. F. Rombauts ◽  
L. A. Salamonsen
Keyword(s):  
Chemokine Receptors ◽  
Embryo Implantation ◽  
Uterine Cavity ◽  
Cycle Phase ◽  
Human Trophoblast ◽  
Endometrial Epithelial Cell ◽  
Glandular Secretion ◽  
Uterine Fluid ◽  
Human Chemokines ◽  
Insight Into

Embryo implantation requires synchronized dialogue between a receptive endometrium and an activated blastocyst via locally produced soluble mediators. During the mid-secretory (MS) phase of the menstrual cycle there is increased glandular secretion into the uterine lumen. These secretions likely contain important mediators that modulate the endometrium and support the conceptus during implantation. Previously we identified that several chemokines were maximally produced during the MS phase by endometrial glandular epithelium (GE) (1, 2) and the presence of chemokine receptors on GE and human trophoblast (3). Furthermore recombinant human chemokines and endometrial epithelial cell-conditioned media stimulated trophoblast migration; this was attenuated by neutralizing specific chemokines (3). Chemokines also regulate a variety of adhesion and ECM molecules on trophoblast (4). Thus chemokines have important roles during embryo implantation. We hypothesized that chemokines are secreted into the uterine cavity and may act on the implanting blastocyst and the endometrium. This study aimed to identify chemokines in uterine fluid (collected by flushing the uterine cavity with 5mls of saline) from fertile women during the proliferative (non-receptive; n=4) and MS (receptive; n=4) phases of the cycle, and from women with unexplained infertility during the MS phase (n=4). Uterine fluid was analyzed using quantitative MilliplexTM Luminex® 42-plex cytokine/chemokine assays revealing the presence of IL-8, CCL2, CCL4, CCL7, CCL11, CCL22 and CX3CL1 in uterine fluid from all women. Importantly chemokine profiles were altered with both cycle phase and fertility; for example CCL4 and CCL22 levels were lower in the infertile cohort, where as CCL2 levels were higher in uterine fluid collected during the proliferative phase. Identifying the soluble mediators in human uterine fluid may provide potential markers of endometrial receptivity, insight into the unique microenvironment essential for pregnancy and a profile of maternal factors that influence the implanting blastocyst.

216. Activin A regulates trophoblast cell adhesion: implications for uterine receptivity and embryo implantation

2008 ◽  
Vol 20 (9) ◽  
pp. 16
Author(s):  
C. J. Stoikos ◽  
A. O.'Connor ◽  
L. A. Salamonsen ◽  
E. Dimitriadis
Keyword(s):  
Embryo Implantation ◽  
Uterine Cavity ◽  
Activin A ◽  
Trophoblast Cell ◽  
Collagen I ◽  
Trophoblast Invasion ◽  
Uterine Receptivity ◽  
Adhesive Properties ◽  
Secretory Phase ◽  
Human Trophoblast

Embryo implantation involves blastocyst attachment to the endometrial luminal epithelium, followed by trophoblast invasion. This process involves a coordinated crosstalk between the implanting blastocyst and the endometrium. Adhesion molecules play an instrumental role during implantation and are regulated by a variety of factors including cytokines and growth factors. Activin A, a TGF-β superfamily member, has been detected in uterine washings,1 and its subunit, β A, is produced by endometrial glands during the secretory phase of the menstrual cycle.2 In endometriosis, a disease that associated with sub-fertility, β A immunostaining is increased in endometrial glands,3 suggesting higher levels of activin A secreted into the uterine lumen could contribute to sub-fertility observed in endometriosis. Therefore we hypothesised that activin A secretion into the uterine cavity affects the adhesive properties of the cells present at the maternal-fetal interface. The aims of the study were to measure and compare activin A secretion in uterine washings from women with and without endometriosis and to demonstrate whether activin A regulates adhesion to extracellular matrix (ECM) components. Uterine washings (5 mL of sterile saline) were collected from women with and without endometriosis during the secretory phase. Activin A was measured by ELISA. HTR8 (human trophoblast cell-line) cells were treated with rhActivin A (50 ng/mL) and assessed for binding to fibronectin, laminin, vitronectin, collagen I and IV. Activin A (>10pg/mL) was detectable in uterine washings from women with and without endometriosis and levels were elevated in endometriosis patients. Untreated HTR8 cells adhered maximally to fibronectin, collagen I and collagen IV with low binding to vitronectin and laminin. Following activin treatment, HTR8 cell binding to fibronectin, collagen I and IV was significantly decreased (n = 3, P < 0.05). These results suggest that activin A regulates the adhesive properties of the blastocyst during implantation. This study also implies that abnormalities in local activin A levels during endometrial receptivity may contribute to sub-fertility in women. (1) Petraglia et., al. (1998) J Clin Endocrinol Metab. (2) Jones et., al. (2000) Mol Hum Reprod. (3) Rombauts et., al. (2006) Aust N Z J Obstet Gynaecol.

scholarly journals A Prospective, Randomized Study of Endometrial Telomerase during the Menstrual Cycle

2001 ◽  
Vol 86 (8) ◽  
pp. 3912-3917 ◽  
Author(s):  
Christopher D. Williams ◽  
John F. Boggess ◽  
L. Robert LaMarque ◽  
William R. Meyer ◽  
Michael J. Murray ◽  
...  
Keyword(s):  
Menstrual Cycle ◽  
Luteal Phase ◽  
Randomized Study ◽  
Embryo Implantation ◽  
Telomerase Activity ◽  
Endometrial Biopsy ◽  
Uterine Receptivity ◽  
Serum Progesterone ◽  
Proliferative Phase ◽  
Late Luteal Phase

The purpose of this study was to characterize telomerase activity during the menstrual cycle, focusing on the luteal phase. A total of 84 endometrial biopsy samples were obtained from 72 participants. Daily urinary LH testing (OvuQuick, Quidel) was used to establish the day of the LH rise, and participants were randomized to return during the secretory phase. Twelve women returned on the identical day during the luteal phase of a subsequent cycle to allow intercycle comparisons of telomerase activity. Telomerase activity was evaluated using a modified TRAP-eze (Intergen) detection protocol. At the time of each endometrial biopsy, serum estrogen and progesterone were measured. Proliferative phase endometrium showed high telomerase activity. At the onset of the luteal phase telomerase activity was high, but it decreased during the early luteal phase, disappeared by the midluteal phase (6 d after LH surge detected), and then rose to moderate levels in the late luteal phase beginning on luteal d 10. Serum progesterone levels were inversely related to telomerase activity. In conclusion, endometrial telomerase activity is dynamic: high during the proliferative phase but inhibited during the midsecretory phase of the menstrual cycle. The timing of expression coincides with the rise and fall of progesterone levels and the time period of maximal uterine receptivity for embryo implantation. This supports a relationship between sex steroid levels and telomerase regulation.

scholarly journals 014.Novel uterine genes in regulation of embryo implantation

2004 ◽  
Vol 16 (9) ◽  
pp. 14
Author(s):  
G. Nie ◽  
Y. Li ◽  
K. Luu ◽  
J. Findlay ◽  
L. Salamonsen
Keyword(s):  
Antisense Oligonucleotides ◽  
Embryo Implantation ◽  
Uterine Receptivity ◽  
Temporal Expression ◽  
Tissue Remodelling ◽  
Functional Studies ◽  
Molecular Events ◽  
Uterine Environment ◽  
Identity Expression

Implantation of the embryo into the maternal endometrium is the first and critical step leading to the establishment of a pregnancy. It has been well established that only during the 'window' of implantation, a limited time span when the uterine environment is receptive, can a blastocyst successfully implant into the uterus. The development of uterine receptivity is accompanied by remarkable morphological and physiological changes in the endometrium, and this is primarily driven by the coordinated effects of the ovarian steroid hormones. Uterine tissue remodelling during implantation also contributes significantly to the development of the placenta. Insufficient uterine remodelling causes implantation failure and infertility. To date, the exact molecular events occurring in the uterus during the establishment of receptivity and at the actual site of implantation are still not well understood. We used the mouse as a model and identified a number of previously unrecognised molecules that are uniquely regulated in the early stages of implantation: one of these is proprotein convertase 6 (PC6). The potential importance of these genes and their products in modulating fertility in the primate, including the human, was demonstrated by their unique spatial and temporal expression in the endometrium of human and rhesus monkey during the phase of uterine receptivity and at implantation. The importance of the genes for implantation was ultimately confirmed by functional studies in vivo using morpholino antisense oligonucleotides. These molecules will be discussed in terms of their identity, expression and functions.

scholarly journals MicroRNAs in small extracellular vesicles indicate successful embryo implantation during early pregnancy

2020 ◽  
Author(s):  
Qiang Tan ◽  
Shuang Shi ◽  
Jingjie Liang ◽  
Xiaowei Zhang ◽  
Dingren Cao ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Early Stage ◽  
Expression Profiles ◽  
Embryo Implantation ◽  
Uterine Cavity ◽  
Synchronous Communication ◽  
Protein Markers ◽  
Uterine Receptivity ◽  
Transmission Electron ◽  
Mirnas Expression

Abstract Background : Synchronous communication between the developing embryo and the receptive endometrium is required for successful embryo implantation. Assessing of uterine receptivity is important for overcoming the recurrent implantation failure (RIF). Although the potential roles of small extracellular vesicles (sEVs) miRNAs in pregnancy have repeatedly mentioned, the systematic study of sEVs derived from endometrium and its cargoes during the implantation have not yet been reported. Methods : In this study, sEVs were isolated from mouse uterine cavity on D2 (pre-receptive phase), D4 (receptive phase) and D5 (implantation) of pregnancy using ultracentrifugation and analyzed by transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). The miRNAs expression profiles of sEVs were identified by RT-qPCR. The datasets analysis of RIF were employed by an integrative bioinformatics analysis. The protein markers of sEVs were examined by western blotting. The effects of miRNAs in embryo implantation were determined by an agomir injection test. Results : Here we show that the number of multivesicular bodies (MVBs) in the endometrium is increased during the window of implantation (WOI). Meanwhile, the expression of CD63 is mainly located in the luminal and glandular epithelium. sEVs miRNAs profile reveal that miR-34c-5p, miR-210, miR-369-5p, miR-30b and miR-582-5p are enriched during WOI. By further integrating the database analysis results of RIF, we found that miR-34c-5p regulates GAS1 to involve normal process of embryo implantation. It is interesting that miR-34c-5p is down-regulated during implantation but enriched in sEVs. An implication of this is the possibility that sEVs miR-34c-5p could be used to evaluate uterine states. Conclusion : Our study identified that mouse endometrium release sEVs during the early stage of pregnancy, especially during WOI. We found that miR-34c-5p in sEVs affects embryo implantation by targeting to GAS1. Furthermore, the sEVs miRNAs suggest potential biomarkers for the choose of suitable period for embryo implantation. This study also has a number of important implications for future practice.

Soluble Delta-like ligand 1 alters human endometrial epithelial cell adhesive capacity

2017 ◽  
Vol 29 (4) ◽  
pp. 694 ◽  
Author(s):  
Michelle Van Sinderen ◽  
Jennifer Oyanedel ◽  
Ellen Menkhorst ◽  
Carly Cuman ◽  
Katarzyna Rainczuk ◽  
...  
Keyword(s):  
Menstrual Cycle ◽  
Embryo Implantation ◽  
Notch Signalling ◽  
Enzyme Linked Immunosorbent Assay ◽  
Secretory Phase ◽  
Functional Changes ◽  
Infertile Women ◽  
Potential Biomarker ◽  
Adhesive Capacity

The endometrium undergoes substantial morphological and functional changes to become receptive to embryo implantation and to enable establishment of a successful pregnancy. Reduced Delta-like ligand 1 (DLL1, Notch ligand) in the endometrium is associated with infertility. DLL1 can be cleaved by ‘a disintegrin and metalloprotease’ (ADAM) proteases to produce a soluble ligand that may act to inhibit Notch signalling. We used an enzyme-linked immunosorbent assay to quantify soluble DLL1 in uterine lavages from fertile and infertile women in the secretory phase of the menstrual cycle. We also determined the cellular location and immunostaining intensity of ADAM12 and 17 in human endometrium throughout the cycle. Functional effects of soluble DLL1 in receptivity were analysed using in vitro adhesion and proliferation assays and gene expression analysis of Notch signalling targets. Soluble DLL1 was significantly increased in uterine lavage samples of infertile women compared with fertile women in the secretory phase of the menstrual cycle. This coincided with significantly increased ADAM17 immunostaining detected in the endometrial luminal epithelium in the mid-secretory phase in infertile women. Soluble DLL1 significantly inhibited the adhesive capacity of endometrial epithelial cells via downregulation of helix–loop–helix and hairy/enhancer of split family member HES1 mRNA. Thus, soluble DLL1 may serve as a suitable target or potential biomarker for receptivity.

scholarly journals A High-Throughput Assay for the Detection of α-Dystroglycan N-Terminus in Human Uterine Fluid to Determine Uterine Receptivity

2015 ◽  
Vol 21 (4) ◽  
pp. 408-413 ◽  
Author(s):  
Sophea Heng ◽  
Beverley Vollenhoven ◽  
Luk J. Rombauts ◽  
Guiying Nie
Keyword(s):  
Embryo Implantation ◽  
Sandwich Elisa ◽  
Hostile Environment ◽  
Uterine Tissue ◽  
Uterine Receptivity ◽  
Biochemical Tests ◽  
Functional Changes ◽  
N Terminus ◽  
Uterine Fluid ◽  
High Throughput Assay

Embryo implantation requires a healthy embryo and a receptive uterus. In women, the uterus remains a hostile environment and must undergo functional changes to convert to a receptive state for embryo implantation. Determining uterine receptivity is vital in IVF treatment, as the timing of embryo transfer needs to be synchronized with uterine receptivity. However, to date, no reliable biochemical tests are available to determine uterine receptivity. We recently established that removal of α-dystroglycan N-terminus (α-DG-N) from the uterine surface plays an important role in the establishment of uterine receptivity. Importantly, the α-DG-N removed from the uterine tissue enters into the uterine fluid, and the levels correlate with the tissue status of receptivity. Detection of α-DG-N in uterine fluid may therefore provide a nonsurgical approach to assess uterine receptivity. In this study, we first validated three monoclonal antibodies raised against α-DG-N in our system, and then established a sandwich ELISA suitable for the detection of α-DG-N in human uterine fluid. This ELISA detected significantly higher concentrations of α-DG-N in uterine fluid of women in the receptive phase. We believe this newly established α-DG-N ELISA may provide an important tool in the development of noninvasive strategies to detect uterine receptivity in women.

Novel aspects of endometrial function: a biological sensor of embryo quality and driver of pregnancy success

2012 ◽  
Vol 24 (1) ◽  
pp. 68 ◽  
Author(s):  
Olivier Sandra ◽  
Nadéra Mansouri-Attia ◽  
Richard G. Lea
Keyword(s):  
Developmental Stages ◽  
Assisted Reproductive Technologies ◽  
Uterine Cavity ◽  
Reproductive Technologies ◽  
Biological Properties ◽  
Reproductive Capacity ◽  
Epigenetic Alterations ◽  
Uterine Receptivity ◽  
Blastocyst Development ◽  
Developmental Potential

Successful pregnancy depends on complex biological processes that are regulated temporally and spatially throughout gestation. The molecular basis of these processes have been examined in relation to gamete quality, early blastocyst development and placental function, and data have been generated showing perturbations of these developmental stages by environmental insults or embryo biotechnologies. The developmental period falling between the entry of the blastocyst into the uterine cavity to implantation has also been examined in terms of the biological function of the endometrium. Indeed several mechanisms underlying uterine receptivity, controlled by maternal factors, and the maternal recognition of pregnancy, requiring conceptus-produced signals, have been clarified. Nevertheless, recent data based on experimental perturbations have unveiled unexpected biological properties of the endometrium (sensor/driver) that make this tissue a dynamic and reactive entity. Persistent or transient modifications in organisation and functionality of the endometrium can dramatically affect pre-implantation embryo trajectory through epigenetic alterations with lasting consequences on later stages of pregnancy, including placentation, fetal development, pregnancy outcome and post-natal health. Developing diagnostic and prognostic tools based on endometrial factors may enable the assessment of maternal reproductive capacity and/or the developmental potential of the embryo, particularly when assisted reproductive technologies are applied.

The Development of the Embryo and Membranes of the Humpback Whale, Megaptera nodosa (Bonnaterre)

1960 ◽  
Vol 11 (3) ◽  
pp. 365 ◽  
Author(s):  
CW Stump ◽  
JP Robins ◽  
ML Garde
Keyword(s):  
Umbilical Cord ◽  
Uterine Cavity ◽  
Distal Region ◽  
Yolk Sac ◽  
Consecutive Series ◽  
Chorionic Villi ◽  
Suspensory Ligament ◽  
Foetal Membranes ◽  
Uterine Fluid ◽  
Tubular Form

The material consists of 20 embryos (5-30 mm) and two foetuses (63 mm and 90 mm) collected at whaling stations on Moreton and Norfolk Islands (latitude 27� 11'S. and 29� 5' S. respectively) during late August, September, and early October in 1952-53-54 and 1956. The consecutive series permitted the study of membrane formation and organogenesis. Younger embryos are found in grooves between the folds of endometrium in a constant site in that uterine horn associated with the ovary containing the recent corpus luteum. Older embryos and the early foetus are adapted to lie freely in the uterine fluid, and are devoid of any mechanism for apposition or attachment to the endometrium. Variation in the sequence of the association of the components of the umbilical cord provides suspensory structures for the amnion and yolk sac, and for the embryo a bifid ligament, retained in the early foetus for attachment of the foetal membranes. In the younger foetus the allantoic duct drains the nephric secretion into the uterine cavity. In the older foetus chorionic villi are present. The bifid suspensory ligament forms the major part of the distal region of the umbilical cord. The allantoic duct is reunited with the allantoic sac. Amniogenesis is by folding. During the embryonic period the chorio-amniotic connection forms a suspensory ligament. The yolk sac, attached by a novel ligament to the amnion, is large and functional in the embryo. In the foetus vascular splanchnopleure is present in a tubular form. A rete system develops in the embryo.

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