Estrogen-induced transcription factor EGR1 regulates c-Kit transcription in the mouse uterus to maintain uterine receptivity for embryo implantation

Abstract To prepare for embryo implantation, the uterus must undergo a series of reciprocal interactions between the uterine epithelium and the underlying stroma, which are orchestrated by ovarian hormones. During this process, multiple signaling pathways are activated to direct cell proliferation and differentiation, which render the uterus receptive to the implanting blastocysts. One important modulator of these signaling pathways is the cell surface and extracellular matrix macromolecules, heparan sulfate proteoglycans (HSPGs). HSPGs play crucial roles in signal transduction by regulating morphogen transport and ligand binding. In this study, we examine the role of HSPG sulfation in regulating uterine receptivity by conditionally deleting the N-deacetylase/N-sulfotransferase (NDST) 1 gene (Ndst1) in the mouse uterus using the Pgr-Cre driver, on an Ndst2- and Ndst3-null genetic background. Although development of the female reproductive tract and subsequent ovarian function appear normal in Ndst triple-knockout females, they are infertile due to implantation defects. Embryo attachment appears to occur but the uterine epithelium at the site of implantation persists rather than disintegrates in the mutant. Uterine epithelial cells continued to proliferate past day 4 of pregnancy, accompanied by elevated Fgf2 and Fgf9 expression, whereas uterine stroma failed to undergo decidualization, as evidenced by lack of Bmp2 induction. Despite normal Indian hedgehog expression, transcripts of Ptch1 and Gli1, both components as well as targets of the hedgehog (Hh) pathway, were detected only in the subepithelial stroma, indicating altered Hh signaling in the mutant uterus. Taken together, these data implicate an essential role for HSPGs in modulating signal transduction during mouse implantation.

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Analysis of heart and neural crest derivatives-expressed protein 2 (HAND2)-progesterone interactions in peri-implantation endometrium†

Biology of Reproduction ◽

10.1093/biolre/ioaa013 ◽

2020 ◽

Vol 102 (5) ◽

pp. 1111-1121 ◽

Cited By ~ 3

Author(s):

Sandra Šućurović ◽

Tamara Nikolić ◽

Jan J Brosens ◽

Biserka Mulac-Jeričević

Keyword(s):

Transcription Factor ◽

Neural Crest ◽

Steroid Hormones ◽

Pregnancy Outcome ◽

Embryo Implantation ◽

Luminal Epithelium ◽

Wild Type ◽

Uterine Receptivity ◽

Ovarian Steroid ◽

Spatiotemporal Analyses

Abstract Implantation is restricted to a narrow window when the local endometrial microenvironment is supportive of the invading embryo. The ovarian steroid hormones estrogen (E) and progesterone (P) are principal regulators of uterine receptivity. Suppression of E-dependent proliferation of luminal epithelium (LE) by P is mandatory for embryo implantation. Here, we report that the balance of E receptor α (ERα) and P receptors (PR) activity controls HAND2 expression, a key transcription factor that determines the fate of the implanting embryo and thereby pregnancy outcome. As a model, we used wild-type mice as well as mice in which either both PR isoforms or the A-isoform was genetically ablated (PRKO and PRAKO, respectively). Detailed spatiotemporal analyses of PR, HAND2, and ERα expression at implantation site demonstrated co-expression of HAND2 and PR but not ERα. Furthermore, in hormonally treated ovariectomized WT, PRAKO and PRKO mice, E suppresses endometrial HAND2 expression. Adding P together with E partially rescues HAND2 expression in WT, but not PRAKO and PRKO animals. Therefore, infertility in PRAKO mice is at least in part associated with the loss of PR-A-regulated HAND2 expression.

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ADAMTS-1: a novel target gene of an estrogen-induced transcription factor, EGR1, critical for embryo implantation in the mouse uterus

Cell & Bioscience ◽

10.1186/s13578-021-00672-8 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Mira Park ◽

So Hee Park ◽

Hyunsun Park ◽

Hye-Ryun Kim ◽

Hyunjung J. Lim ◽

...

Keyword(s):

Transcription Factor ◽

Target Gene ◽

Target Genes ◽

Expression Profiles ◽

Embryo Implantation ◽

Uterine Epithelium ◽

Expression Vectors ◽

Dependent Manner ◽

Mouse Uterus ◽

Novel Target

Abstract Background Recently, we demonstrated that estrogen (E2) induces early growth response 1 (Egr1) to mediate its actions on the uterine epithelium by controlling progesterone receptor signaling for successful embryo implantation. EGR1 is a transcription factor that regulates the spectrum of target genes in many different tissues, including the uterus. E2-induced EGR1 regulates a set of genes involved in epithelial cell remodeling during embryo implantation in the uterus. However, only few target genes of EGR1 in the uterus have been identified. Result The expression of ADAM metallopeptidase with thrombospondin type 1 motif 1 (Adamts-1) was significantly downregulated in the uteri of E2-treated ovariectomized (OVX) Egr1(−/−) mice. Immunostaining of ADAMTS-1 revealed its exclusive expression in the uterine epithelium of OVX wild-type but not Egr1(−/−) mice treated with E2. The expression profiles of Adamts-1 and Egr1 were similar in the uteri of E2-treated OVX mice at various time points tested. Pre-treatment with ICI 182, 780, a nuclear estrogen receptor (ER) antagonist, effectively inhibited the E2-dependent induction of Egr1 and Adamts-1. Pharmacologic inhibition of E2-induced ERK1/2 or p38 phosphorylation interfered with the induction of EGR1 and ADAMTS-1. Furthermore, ADAMTS-1, as well as EGR1, was induced in stroma cells surrounding the implanting blastocyst during embryo implantation. Transient transfection with EGR1 expression vectors significantly induced the expression of ADAMTS-1. Luciferase activity of the Adamts-1 promoter containing EGR1 binding sites (EBSs) was increased by EGR1 in a dose-dependent manner, suggesting functional regulation of Adamts-1 transcription by EGR1. Site-directed mutagenesis of EBS on the Adamts-1 promoter demonstrated that EGR1 directly binds to the EBS at -1151/-1134 among four putative EBSs. Conclusions Collectively, we have demonstrated that Adamts-1 is a novel target gene of E2-ER-MAPK-EGR1, which is critical for embryo implantation in the mouse uterus during early pregnancy.

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Changes in anandamide levels in mouse uterus are associated with uterine receptivity for embryo implantation

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.94.8.4188 ◽

1997 ◽

Vol 94 (8) ◽

pp. 4188-4192 ◽

Cited By ~ 182

Author(s):

P. C. Schmid ◽

B. C. Paria ◽

R. J. Krebsbach ◽

H. H. O. Schmid ◽

S. K. Dey

Keyword(s):

Embryo Implantation ◽

Uterine Receptivity ◽

Mouse Uterus

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The Notch Family Transcription Factor, RBPJκ, Modulates Glucose Transporter and Ovarian Steroid Hormone Receptor Expression During Decidualization

Reproductive Sciences ◽

10.1177/1933719118799209 ◽

2018 ◽

Vol 26 (6) ◽

pp. 774-784 ◽

Cited By ~ 1

Author(s):

Michael R Strug ◽

Ren-Wei Su ◽

Tae Hoon Kim ◽

Jae-Wook Jeong ◽

Asgerally Fazleabas

Keyword(s):

Transcription Factor ◽

Notch Signaling ◽

Glucose Transporter ◽

Embryo Implantation ◽

Receptor Expression ◽

Marker Genes ◽

Ovarian Steroid ◽

Mouse Uterus ◽

Endometrial Stromal Cells

During decidualization, endometrial stromal cells differentiate into a secretory phenotype to modulate the uterine microenvironment and promote embryo implantation. This highly metabolic process relies on ovarian steroid receptors and glucose transporters. Canonical Notch signaling is mediated by the transcription factor Recombination Signal Binding Protein for Immunoglobulin Kappa J Region (RBPJ). Loss of RBPJ in the mouse uterus ( Pgrcre/+Rbpjflox/flox; Rbpj c-KO) results in subfertility in part due to an abnormal uterine–embryonic axis during implantation and, as described herein, decidualization failure. Induced in vivo decidualization in Rbpj c-KO mice was impaired with the downregulation of decidual markers and decreased progesterone receptor (Pgr) signaling. Consistent with in vivo mouse data, RBPJ knockdown during in vitro Human uterine fibroblast (HuF) cell decidualization results in the reduced expression of decidual marker genes along with PGR. Expression of the glucose transporter, SLC2A1, was decreased in the RBPJ-silenced HuF cells, which corresponded to decreased Slc2a1 in the secondary decidual zone of Rbpj c-KO mouse uteri. Exogenous administration of pyruvate, which bypasses the need for glucose, rescues PRL expression in RBPJ-deficient HuF cells. In summary, Notch signaling through RBPJ controls both ovarian steroid receptor PGR and glucose transporter SLC2A1 expression during decidualization, and this dysregulation likely contributes to embryo implantation failure.

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Nuclear Shp2 directs normal embryo implantation via facilitating the ERα tyrosine phosphorylation by the Src kinase

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1700978114 ◽

2017 ◽

Vol 114 (18) ◽

pp. 4816-4821 ◽

Cited By ~ 11

Author(s):

Hao Ran ◽

Shuangbo Kong ◽

Shuang Zhang ◽

Jianghong Cheng ◽

Chan Zhou ◽

...

Keyword(s):

Tyrosine Phosphorylation ◽

Tyrosine Phosphatase ◽

Embryo Implantation ◽

Src Kinase ◽

Estrogen Receptor Α ◽

Normal Embryo ◽

Uterine Receptivity ◽

Mouse Uterus ◽

Independent Manner ◽

Rtk Signaling

Estrogen and progesterone coupled with locally produced signaling molecules are essential for embryo implantation. However, the hierarchical landscape of the molecular pathways that governs this process remains largely unexplored. Here we show that the protein tyrosine phosphatase Shp2, a positive transducer of RTK signaling, is predominately localized in the nuclei in the periimplantation mouse uterus. Uterine-specific deletion of Shp2 exhibits reduced progesterone receptor (PR) expression and progesterone resistance, which derails normal uterine receptivity, leading to complete implantation failure in mice. Notably, the PR expression defects are attributed to the limited estrogen receptor α (ERα) activation in uterine stroma. Further analysis reveals that nuclear Shp2, rather than cytosolic Shp2, promotes the ERα transcription activity. This function is achieved by enhancing the Src kinase-mediated ERα tyrosine phosphorylation, which facilitates ERα binding to Pgr promoter in an ERK-independent manner in periimplantation uteri. Besides uncovering a regulatory mechanism, this study could be clinically relevant to dysfunctional ERα-caused endometrial disorders in women.

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Inhibition of TMEM16A impedes embryo implantation and decidualization in mice

Reproduction ◽

10.1530/rep-18-0197 ◽

2018 ◽

Cited By ~ 2

Author(s):

Qianrong Qi ◽

Yifan Yang ◽

Kailin Wu ◽

Qingzhen Xie

Keyword(s):

Progesterone Receptor ◽

Stromal Cells ◽

Embryo Implantation ◽

Mouse Uterus ◽

Endometrial Stromal Cells ◽

Uterine Endometrium ◽

Molecule Inhibitor ◽

Pathway Inhibition ◽

And Function ◽

Reproductive Processes

Recent studies revealed that TMEM16A is involved in several reproductive processes, including ovarian estrogen secretion and ovulation, sperm motility and acrosome reaction, fertilization, and myometrium contraction. However, little is known about the expression and function of TMEM16A in embryo implantation and decidualization. In this study, we focused on the expression and regulation of TMEM16A in mouse uterus during early pregnancy. We found that TMEM16A is up-regulated in uterine endometrium in response to embryo implantation and decidualization. Progesterone treatment could induce TMEM16A expression in endometrial stromal cells through progesterone receptor/c-Myc pathway, which is blocked by progesterone receptor antagonist or the inhibitor of c-Myc signaling pathway. Inhibition of TMEM16A by small molecule inhibitor (T16Ainh-A01) resulted in impaired embryo implantation and decidualization in mice. Treatment with either specific siRNA of Tmem16a or T16Ainh-A01 inhibited the decidualization and proliferation of mouse endometrial stromal cells. In conclusion, our results revealed that TMEM16A is involved in embryo implantation and decidualization in mice, compromised function of TMEM16A may lead to impaired embryo implantation and decidualization.

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Cell–Cell Communication at the Embryo Implantation Site of Mouse Uterus Revealed by Single-Cell Analysis

International Journal of Molecular Sciences ◽

10.3390/ijms22105177 ◽

2021 ◽

Vol 22 (10) ◽

pp. 5177

Author(s):

Yi Yang ◽

Jia-Peng He ◽

Ji-Long Liu

Keyword(s):

Single Cell ◽

Molecular Mechanism ◽

Single Cell Analysis ◽

Embryo Implantation ◽

Cell Communication ◽

Human Reproduction ◽

Mouse Uterus ◽

Implantation Sites ◽

Low Efficiency ◽

Signaling Interactions

As a crucial step for human reproduction, embryo implantation is a low-efficiency process. Despite rapid advances in recent years, the molecular mechanism underlying embryo implantation remains poorly understood. Here, we used the mouse as an animal model and generated a single-cell transcriptomic atlas of embryo implantation sites. By analyzing inter-implantation sites of the uterus as control, we were able to identify global gene expression changes associated with embryo implantation in each cell type. Additionally, we predicted signaling interactions between uterine luminal epithelial cells and mural trophectoderm of blastocysts, which represent the key mechanism of embryo implantation. We also predicted signaling interactions between uterine epithelial-stromal crosstalk at implantation sites, which are crucial for post-implantation development. Our data provide a valuable resource for deciphering the molecular mechanism underlying embryo implantation.

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Sperm modulate uterine immune parameters relevant to embryo implantation and reproductive success in mice

Communications Biology ◽

10.1038/s42003-021-02038-9 ◽

2021 ◽

Vol 4 (1) ◽

Author(s):

John E. Schjenken ◽

David J. Sharkey ◽

Ella S. Green ◽

Hon Yeung Chan ◽

Ricky A. Matias ◽

...

Keyword(s):

Reproductive Success ◽

Seminal Fluid ◽

Embryo Implantation ◽

Reproductive Investment ◽

Global Gene Expression ◽

Wild Type ◽

Mouse Uterus ◽

Immune Parameters ◽

Oocyte Fertilization

AbstractSeminal fluid factors modulate the female immune response at conception to facilitate embryo implantation and reproductive success. Whether sperm affect this response has not been clear. We evaluated global gene expression by microarray in the mouse uterus after mating with intact or vasectomized males. Intact males induced greater changes in gene transcription, prominently affecting pro-inflammatory cytokine and immune regulatory genes, with TLR4 signaling identified as a top-ranked upstream driver. Recruitment of neutrophils and expansion of peripheral regulatory T cells were elevated by seminal fluid of intact males. In vitro, epididymal sperm induced IL6, CXCL2, and CSF3 in uterine epithelial cells of wild-type, but not Tlr4 null females. Collectively these experiments show that sperm assist in promoting female immune tolerance by eliciting uterine cytokine expression through TLR4-dependent signaling. The findings indicate a biological role for sperm beyond oocyte fertilization, in modulating immune mechanisms involved in female control of reproductive investment.

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COUP-TFII Regulates Human Endometrial Stromal Genes Involved in Inflammation

Molecular Endocrinology ◽

10.1210/me.2013-1191 ◽

2013 ◽

Vol 27 (12) ◽

pp. 2041-2054 ◽

Cited By ~ 32

Author(s):

Xilong Li ◽

Michael J. Large ◽

Chad J. Creighton ◽

Rainer B. Lanz ◽

Jae-Wook Jeong ◽

...

Keyword(s):

Cell Fate ◽

Cell Biology ◽

Embryo Implantation ◽

Orphan Nuclear Receptor ◽

Loss Of Function ◽

Mouse Uterus ◽

Endometrial Stromal Cells ◽

Endometrial Stroma ◽

Stroma Cell

Chicken ovalbumin upstream promoter-transcription factor II (COUP-TFII; NR2F2) is an orphan nuclear receptor involved in cell-fate specification, organogenesis, angiogenesis, and metabolism. Ablation of COUP-TFII in the mouse uterus causes infertility due to defects in embryo attachment and impaired uterine stromal cell decidualization. Although the function of COUP-TFII in uterine decidualization has been described in mice, its role in the human uterus remains unknown. We observed that, as in mice, COUP-TFII is robustly expressed in the endometrial stroma of healthy women, and its expression is reduced in the ectopic lesions of women with endometriosis. To interrogate the role of COUP-TFII in human endometrial function, we used a small interfering RNA-mediated loss of function approach in primary human endometrial stromal cells. Attenuation of COUP-TFII expression did not completely block decidualization; rather it had a selective effect on gene expression. To better elucidate the role of COUP-TFII in endometrial stroma cell biology, the COUP-TFII transcriptome was defined by pairing microarray comparison with chromatin immunoprecipitation followed by deep sequencing. Gene ontology analysis demonstrates that COUP-TFII regulates a subset of genes in endometrial stroma cell decidualization such as those involved in cell adhesion, angiogenesis, and inflammation. Importantly this analysis shows that COUP-TFII plays a role in controlling the expression of inflammatory cytokines. The determination that COUP-TFII plays a role in inflammation may add insight into the role of COUP-TFII in embryo implantation and in endometrial diseases such as endometriosis.

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