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The process of embryo attachment and invasion through the endometrial epithelial cells and subsequent implantation into the decidualized endometrial stroma is the groundbreaking step for the establishment of a successful pregnancy. Necessary prerequisites are a receptive endometrium, a good-quality embryo and a well-orchestrated molecular dialog between embryo and maternal endometrium. The embryo-maternal dialog is conducted via a wide scope of factors, including secreted cytokines, chemokines, and growth factors in addition to the expression of corresponding receptors and co-receptors. Several embryonic proteins, including the aforementioned, are involved in the process of apoptosis, which necessarily needs to take place at the maternal endometrium to allow the embryo to invade. The endometrial epithelium is thereby disintegrated completely within a particular area, whereas the endometrial stroma seems to require a more depth-limited apoptosis. As of today, the exact mechanisms and factors mediating the apoptotic process involved in those apparently differently regulated incidents are not fully understood, particularly with regard to stromal cell apoptosis. There is evidence though, that cytokines and their respective receptors play a major role. A suggested important co-receptor for cytokines, which is highly upregulated in the receptive human endometrium, is the heparan sulfate proteoglycan syndecan-1. It is present on the cell surface and involved in the regulation of cell-cell-interaction, cell binding, cell signaling and cytoskeletal organization and therefore represents a possible mediator of apoptosis regulation in human endometrium. Herein, the literature on endometrial epithelial and stromal apoptosis in general, and in light of the influence of syndecan-1, is reviewed.

Human chorionic gonadotropin (hCG) has long been associated with the initiation and maintenance of pregnancy, where angiogenesis plays an important role. However, the function of hCG in angiogenesis and the recruitment of vascular active cells are not fully understood. In this study, the role of hCG and its receptor in circulating angiogenic and human endothelial cells, including lymphatic, uterine microvascular, and umbilical vein endothelial cells, was examined. Immunohistochemistry and immunoblot analysis were used to detect LH/hCG receptor expression and the expression of hCG-induced angiogenic molecules. HIF-1α was determined via ELISA and downstream molecules, such as CXCL12 and CXCR4, via real-time PCR. Chemotaxis was analyzed using Boyden chambers. Our results show that the LH/hCG receptor was present in all tested cells. Furthermore, hCG was able to stimulate LH/hCG-receptor-specific migration in a dose-dependent fashion and induce key angiogenic molecules, including HIF-1α, CXCL12, and CXCR4. In conclusion, our findings underscore the importance of hCG as one of the first angiogenic molecules produced by the conceptus. hCG itself alters endothelial motility, recruitment, and expression of pro-angiogenic molecules and may therefore play an important role in vascular adaption during implantation and early placental formation.

The placenta forms the interface between the mother and the fetus. During placental development cytotrophoblasts differentiate to form the syncytium or to invade the decidual wall to breach maternal vessels and establish the blood flow in the intervillous space. This process is still not well understood but it is proposed that chemokines and their receptors are involved in guiding cytotrophoblasts to the decidua and maternal vessels as well as attracting immunocompetent cells to the implantation site. CXCL12 is a chemokine expressed by cytotrophoblasts and is involved in cytotrophoblast invasion, differentiation and survival. One of its receptors, CXCR4, has been detected on cytotrophoblasts. Recent data show that CXCR7 and syndecan-4 might partially mediate CXCL12 function in other cell types. In this study, we examined CXCR7 and syndecan-4 expression at the maternal-fetal interface via immmunolocalization in placental tissue sections and in isolated cytotrophoblasts. We further used immunoblot analyses to confirm the data. We were able to show that cytotrophoblasts express both receptors and that upregulation occurs during the differentiation process of cytotrophoblasts towards the invasive phenotype. On a functional level CXCR7 seems not to be involved in JAR cell chemotaxis, suggesting a different function of this receptor. In conclusion, we propose that CXCL12 binds to CXCR4, but also to CXCR7 and syndecan-4. These three receptors could mediate different functions of CXCL12, such as cell migration, directed invasion, proliferation and survival. The latter molecules might also be involved in the development of placental pathologies, such as preeclampsia or choriocarcinoma growth.

Angiogenesis is a key process in the endometrium which undergoes dramatic changes during the menstrual cycle. Molecules such as vascular endothelial growth factor (VEGF), acting via two tyrosine kinase family receptors (VEGFR1 [Flt-1] and VEGFR2 [KDR/Flk-1]), are potent modulators of angiogenesis and vascular remodelling in the endometrium. Recently, neuropilin-1 (NRP-1) was shown to be expressed in endothelial cells binding VEGF(165) and therewith enhancing the binding of VEGF(165) to VEGFR2. This suggests that NRP-1, in addition to the known VEGF receptors, may play an important role in VEGF-induced angiogenesis. In this study, the expression of NRP-1 in the cycling human endometrium has been investigated by reverse transcription (RT)-polymerase chain reaction (RT-PCR), semi-quantitative competitive RT-PCR (RT-cPCR) and immunohistochemical staining. NRP-1 was expressed in all 32 endometrium samples throughout the menstrual cycle. However, samples from the proliferative phase showed significantly higher expression levels of NRP-1 mRNA compared to samples from the secretory phase (t/c-ratio 2.13 vs. 0.84, p0.05). Immunohistochemistry confirmed the results showing increased NRP-1 staining in vascular endothelium, glandular epithelium and stromal cells of the proliferative phase endometrium. This study demonstrates mRNA and protein expression of NRP-1 in human endometrium samples throughout the menstrual cycle. The enhanced expression of NRP-1 in the proliferative phase suggests that it may participate in hormonally regulated changes of endometrial angiogenesis, preparing the endometrium for the implantation of an embryo. NRP-1 expression might act as a co-factor for VEGF(165) enhancing the angiogenic stimulus.

The follicle provides a unique micro-environment for the oocyte to develop and mature to a fertilizable gamete, within. Intrafollicular components, such as follicular fluid and cellular material are a potential resource for the noninvasive prediction of oocyte developmental potential. Using the cow as a model, we have established in vitro embryo production (IVP) methods that facilitate the individual tracking of oocytes and embryos, permitting a retrospective analysis of retained follicular components according to thedevelopmental outcomeof the oocyte during IVP. We have investigated the relationship between a range of parameters (including intrafollicular steroids and metabolomic profiles in follicle fluid and gene expression in follicular and cumulus cells) and oocyte development in vitro. Intrafollicular steroid concentrations in fluid from 3to 8-mm antral follicles were not predictive of oocyte developmental competence. However, metabolomic profiling of the same follicular fluid was highly predictive; several amino acids and polyunsaturated fatty acids positively correlated with blastocyst formation. Metabolomic analyses of follicular fluid from alternative bovine models of fertility have confirmed the predictive value of this approach. Employing both candidate and global gene expression analysis of transcript abundance in follicular and cumulus cells, we identified differentially expressed transcripts that were positively associated with blastocyst formation. However, many candidate genes are only predictivewithin themodel inwhich theywerefirst identified. Recently, we have focused on the role of progesterone (P4) during oocyte maturation. P4 has an anti-apoptotic effect on the cumulus–oocyte complex during maturation that is mediated via the nuclear P4 receptors. We have identified P4-responsive genes that have similar expression profiles in both the oocyte and their cumulus cells, implying their potential as biomarkers of oocyte quality. In particular, the pro-survival and cell cycle factor AVEN, whose dynamic regulation by P4 is reflected in both the oocyte and surrounding cumulus cells could potentially be exploited not just as an indicatorof oocytequality, but also as an indicator of IVCconditions. In summary, follicular components are an excellent resource for thenon-invasivepredictionofoocyte developmental potential; however, not all parameters are predictive across all models of oocyte development.