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Syndecan-1 knock-down in decidualized human endometrial stromal cells leads to significant changes in cytokine and angiogenic factor expression patterns

BackgroundSuccessful embryonic implantation depends on a synchronized embryo-maternal dialogue. Chemokines, such as chemokine ligand 1 (CXCL1), play essential roles in the maternal reproductive tract leading to morphological changes during decidualization, mediating maternal acceptance towards the semi-allograft embryo and induction of angiogenesis. Chemokine binding to their classical G-protein coupled receptors is essentially supported by the syndecan (Sdc) family of heparan sulfate proteoglycans. The aim of this study was to identify the involvement of Sdc-1 at the embryo-maternal interface regarding changes of the chemokine and angiogenic profile of the decidua during the process of decidualization and implantation in human endometrium.MethodsA stable Sdc-1 knock-down was generated in the immortalized human endometrial stromal cell line St-T1 and was named KdS1. The ability of KdS1 to decidualize was proven by Insulin-like growth factor binding 1 (IGFBP1) and prolactin (PRL) confirmation on mRNA level before further experiments were carried out. Dot blot protein analyses of decidualized knock-down cells vs non-transfected controls were performed. In order to imitate embryonic implantation, decidualized KdS1 were then incubated with IL-1beta, an embryo secretion product, vs controls. Statistical analyses were performed applying the Students t-test with p < 0.05, p < 0.02 and p < 0.01 and one way post-hoc ANOVA test with p < 0.05 as cut-offs for statistical significance.ResultsThe induction of the Sdc-1 knock-down revealed significant changes in cytokine and angiogenic factor expression profiles of dKdS1 vs decidualized controls. Incubation with embryonic IL-1beta altered the expression patterns of KdS1 chemokines and angiogenic factors towards inflammatory-associated molecules and factors involved in matrix regulation.ConclusionsSdc-1 knock-down in human endometrial stroma cells led to fulminant changes regarding cytokine and angiogenic factor expression profiles upon decidualization and imitation of embryonic contact. Sdc-1 appears to play an important role as a co-receptor and storage factor for many cytokines and angiogenic factors during decidualization and implantation period, supporting proper implantation and angiogenesis by regulation of chemokine and angiogenic factor secretion in favour of the implanting embryo.

CXCR7 and syndecan-4 are potential receptors for CXCL12 in human cytotrophoblasts

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.

Angiopoietin-1 and -2 mRNA and protein expression in mouse preimplantation embryos and uteri suggests a role in angiogenesis during implantation.

After attachment and migration through the endometrial epithelium, the embryo must induce angiogenesis within the endometrial stroma to successfully complete the implantation process. Growth factors have been shown to play an important role in embryo implantation and placentation. The aim of the study was to investigate the expression of angiopoietin-1 and -2 (Ang-1 and -2) mRNA and protein expression during the development of single preimplantation mouse embryos and of possible complementary expression in mouse uteri. Angiopoietin-1 mRNA was expressed throughout development in 78% of zygotes, 66% of 2-cell-embryos, 71% of 4-cell-embryos, 70% of 8-cell-embryos, 60% of morula stages, 48% of early blastocysts and 78% of late blastocysts. The number of Ang-1-expressing embryos in the early-blastocyst group was significantly different in comparison with zygotes, 4-cell-embryos, 8-cell-embryos and late blastocysts. Angiopoietin-2 mRNA and protein expression could not be detected in preimplantation embryos. Examination of the uteri revealed Ang-2 mRNA and protein expression in the oestrogen-dominated cycling phase and the progesterone-dominated mated phase, whereas Ang-1 expression was restricted to the mated phase. Herein, Ang-1 expression in preimplantation mouse embryos as well as Ang-1 and -2 expression in mouse uteri is demonstrated, suggesting a possible role for angiopoietins in the embryo-maternal dialogue of the implantation process via an enhancement of the vascular remodelling in favour of an implanting conceptus.

Molekulare Mechanismen der Embryoimplantation im Endometrium

ZusammenfassungDie menschliche Reproduktion und die Implantation des Embryos sind ein komplexer und dadurch störungsanfälliger Prozess. Ein genauer Zeitplan bestimmt die Synchronisation der Blastozystenreifung und die endometriale Vorbereitung auf die Einnistung. Eine Vertiefung unseres Wissens über die molekularen Abläufe während der physiologischen Präimplantationsentwicklung und Implantation sollte zu einer Verbesserung der In-vitro-Kulturbedingungen und damit ultimativ zu einer Steigerung der Erfolgsraten der In-vitro-Fertilisationsbehandlung führen. In dem vorliegenden Beitrag werden molekulare Mechanismen, wie hormonelle Faktoren, Zytokin- und Wachstumsfaktorensysteme, die Bedeutung von Metalloproteinasen und Adhäsionsmolekülen kurz beschrieben und in den Kontext aktueller Forschung gestellt. Die weitere Erforschung dieser Zusammenhänge wird verantwortlich dafür sein, dass in Zukunft ein besseres klinisches sowie therapeutisches Management im Hinblick auf Sterilität, Infertilität, jedoch auch Kontrazeption entwickelt werden kann.AbstractHuman Reproduction is a very complex and surprisingly inefficient process. The embryo development and the generation of a receptive endometrium needs an exact timing and synchronization. Understanding the factors involved in preimplantation embryo development and embryo-maternal interaction and implantation is crucial for improving success rates in reproductive medicine. The purpose of the present review is to describe briefly the function of adhesion- and hormonal factors, cytokines, growth factors and metalloproteinases. A better understanding of the molecular mechanisms of implantation will be responsible for the improvement of clinical and therapeutical management of sterility, infertility and contraception.

Syndecan-1 Acts as an Important Regulator of CXCL1 Expression and Cellular Interaction of Human Endometrial Stromal and Trophoblast Cells

Successful implantation of the embryo into the human receptive endometrium is substantial for the establishment of a healthy pregnancy. This study focusses on the role of Syndecan-1 at the embryo-maternal interface, the multitasking coreceptor influencing ligand concentration, release and receptor presentation, and cellular morphology. CXC motif ligand 1, being involved in chemotaxis and angiogenesis during implantation, is of special interest as a ligand of Syndecan-1. Human endometrial stromal cells with and without Syndecan-1 knock-down were decidualized and treated with specific inhibitors to evaluate signaling pathways regulating CXC ligand 1 expression. Western blot analyses of MAPK and Wnt members were performed, followed by analysis of spheroid interactions between human endometrial cells and extravillous trophoblast cells. By mimicking embryo contact using IL-1β, we showed less ERK and c-Jun activation by depletion of Syndecan-1 and less Frizzled 4 production as part of the canonical Wnt pathway. Additionally, more beta-catenin was phosphorylated and therefore degraded after depletion of Syndecan-1. Secretion of CXC motif ligand 1 depends on MEK-1 with respect to Syndecan-1. Regarding the interaction of endometrial and trophoblast cells, the spheroid center-to-center distances were smaller after depletion of Syndecan-1. Therefore, Syndecan-1 seems to affect signaling processes relevant to signaling and intercellular interaction at the trophoblast-decidual interface.

Wann ist die Chance auf Mutterschaft vertan

Viele Frauen, die die Familienplanung immer weiter hinausschieben, verdrängen die Tatsache, dass ihre Fertilität etwa ab dem 35. Lebensjahr drastisch sinkt. Was heute über die „Biologie des reproduktiven Alters“ bekannt ist und mit welchen Methoden Frauen im kritischen Alter doch noch zu einem eigenen Kind verholfen werden kann, ist Thema des folgenden Beitrags.

Rettung vor der Infertilität im Alter

Über Social Freezing, dem vorsorglichen Einfrieren von Eizellen, damit eine Frau auch noch in späteren Jahren schwanger werden kann, wird viel diskutiert. Im Mittelpunkt des nachfolgenden Beitrags stehen weniger ethische als vielmehr medizinische und gesellschaftliche Aspekte. Wie funktioniert Sozial Freezing, welche Erfolgsraten sind zu erwarten, wer lässt sich behandeln, und was kostet die Behandlung?