scholarly journals Differential roles of uterine epithelial and stromal STAT3 coordinate uterine receptivity and embryo attachment

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Takehiro Hiraoka ◽  
Yasushi Hirota ◽  
Yamato Fukui ◽  
Mona Gebril ◽  
Tetsuaki Kaku ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Embryo Implantation ◽  
Uterine Receptivity ◽  
Uterine Lumen ◽  
Female Mice ◽  
Attachment Sites ◽  
Transcription 3 ◽  
Estrogenic Responses ◽  
Embryo Attachment

Abstract Although it has been reported that uterine signal transducer and activator of transcription 3 (STAT3) is essential for embryo implantation, the exact roles of uterine epithelial and stromal STAT3 on embryo implantation have not been elucidated. To address this issue, we generated Stat3-floxed/Ltf-iCre (Stat3-eKO), Stat3-floxed/Amhr2-Cre (Stat3-sKO), and Stat3-floxed/Pgr-Cre (Stat3-uKO) mice to delete Stat3 in uterine epithelium, uterine stroma, and whole uterine layers, respectively. We found that both epithelial and stromal STAT3 have critical roles in embryo attachment because all the Stat3-eKO and Stat3-sKO female mice were infertile due to implantation failure without any embryo attachment sites. Stat3-eKO uteri showed indented structure of uterine lumen, indicating the role of epithelial STAT3 in slit-like lumen formation in the peri-implantation uterus. Stat3-sKO uteri exhibited hyper-estrogenic responses and persistent cell proliferation of the epithelium in the peri-implantation uterus, suggesting the role of stromal STAT3 in uterine receptivity. In addition, Stat3-uKO female mice possessed not only the characteristic of persistent epithelial proliferation but also that of indented structure of uterine lumen. These findings indicate that epithelial STAT3 controls the formation of slit-like structure in uterine lumen and stromal STAT3 suppresses epithelial estrogenic responses and cell proliferation. Thus, epithelial and stromal STAT3 cooperatively controls uterine receptivity and embryo attachment through their different pathways.

scholarly journals Influence of hyaluronan on endometrial receptivity and embryo attachment in sheep

2017 ◽  
Vol 29 (9) ◽  
pp. 1763 ◽  
Author(s):  
Waleed F. A. Marei ◽  
D. Claire Wathes ◽  
Kabir A. Raheem ◽  
Omnia Mohey-Elsaeed ◽  
Fataneh Ghafari ◽  
...  
Keyword(s):  
Embryo Implantation ◽  
Endometrial Receptivity ◽  
Uterine Receptivity ◽  
Female Reproduction ◽  
Mucin 1 ◽  
Synthesis Inhibitor ◽  
The Impact ◽  
Hyaluronidase 2 ◽  
Embryo Attachment

An increasing number of reports suggests a role of hyaluronan (HA) in female reproduction and interest in its application in assisted reproduction is rising. However, there are contrasting data about the effectiveness of adding HA to the embryo-transfer medium on improving pregnancy rates. Using sheep as an experimental model, the studies reported here analysed the impact of HA infusion into the uterus on embryo attachment to uterine luminal epithelium (LE) and expression of selected markers of uterine receptivity. On Day 14 after natural mating (pre-attachment), uterine horns were infused with either (n = 4 each): PBS (control), HA (1 mg mL–1), HA + hyaluronidase 2 (Hyal2; 300 IU mL–1) or 4-methyl-umbelliferone (HA-synthesis inhibitor; 4MU, 1 mM). HA immunostaining on uterine sections collected on Day 17 was negative in the 4MU group and weak in the HA+Hyal2 group. In contrast to 4MU, which resulted in 100% attachment, HA infusion blocked embryo attachment in all treated animals. This was accompanied by the disappearance of mucin 1 and increased expression of osteopontin and CD44v6 in the LE of uteri with attached embryos. In conclusion, the presence of HA at the embryo–maternal interface during embryo implantation resulted in reduced endometrial receptivity and inhibited the interaction of trophoblasts with the LE, whereas clearance of HA favoured embryo attachment.

Levonorgestrel Inhibits Embryo Attachment by Eliminating Uterine Induction of Leukemia Inhibitory Factor

Endocrinology ◽  
2019 ◽  
Vol 161 (2) ◽  
Author(s):  
Mitsunori Matsuo ◽  
Yasushi Hirota ◽  
Yamato Fukui ◽  
Hidetoshi Fujita ◽  
Tomoko Saito-Fujita ◽  
...  
Keyword(s):  
Leukemia Inhibitory Factor ◽  
Embryo Implantation ◽  
Infertility Treatment ◽  
Vehicle Control ◽  
Inhibitory Factor ◽  
Wild Type ◽  
Attachment Sites ◽  
Attachment Inhibition ◽  
Evening Administration ◽  
Embryo Attachment

Abstract Progestogens including progesterone (P4) and levonorgestrel (LNG) are clinically used for multiple purposes such as contraception and infertility treatment. The effects of progestogens on the uterus remains to be elucidated. Here we examine the effect of excessive progestogen administration on embryo implantation focusing on the function of uterine leukemia inhibitory factor (LIF), a cytokine that is induced by estrogen and essential for embryo attachment. Treatment of wild-type (WT) female mice with vehicle (control), LNG at the dose of 300 μg/kg/day and P4 at the dose of 10 mg/day from day 1 to day 4 of pregnancy was conducted. LNG-treated and P4-treated mice showed embryo attachment failure on day 5 of pregnancy (The rate of mice with embryo attachment sites [%MAS], 11% and 13%, respectively), while all the control mice had normal attachment sites. Uterine LIF expression was significantly reduced in LNG-treated and P4-treated mice on day 4 evening. Administration of recombinant LIF (rLIF) at the dose of 24 μg/day on day 4 significantly rescued embryo attachment failure in LNG-treated and P4-treated mice (%MAS, 80% and 75%, respectively). Estradiol (E2) administration also rescued embryo attachment failure in LNG-treated mice (%MAS, 83%). Furthermore, excess P4 treatment before implantation decreased decidual P4 receptor (PGR) expression and induced decidualization defect apart from LIF downregulation. These findings indicate that progestogens cause embryo attachment inhibition through downregulation of uterine LIF expression and compromised decidualization through downregulation of PGR independently of LIF reduction. This study may contribute to a better understanding of contraceptive action of progestogens.

scholarly journals MiR199a is implicated in embryo implantation by regulating Grb10 in rat

Reproduction ◽  
2014 ◽  
Vol 147 (1) ◽  
pp. 91-99 ◽  
Author(s):  
Hong-Fei Xia ◽  
Jing-Li Cao ◽  
Xiao-Hua Jin ◽  
Xu Ma
Keyword(s):  
Cell Proliferation ◽  
Embryo Implantation ◽  
Growth Factor Receptor ◽  
Inverse Association ◽  
Loss Of Function ◽  
Endometrial Stromal Cells ◽  
Proliferation And Apoptosis ◽  
17Β Estradiol

MiR199a was found to be differentially expressed in rat uteri between the prereceptive and receptive phase via microRNA (miRNA) microarray analysis in our previous study. However, the role of miR199a in rat embryo implantation remained unknown. In the study, northern blot results showed that the expression levels of miR199a were higher on gestation days 5 and 6 (g.d.5–6) in rat uteri than on g.d.3–4 and g.d.7–8. In situ localization of miR199a in rat uteri showed that miR199a was mainly localized in the stroma or decidua. The expression of miR199a was not significantly different in the uteri of pseudopregnant rats and evidently increased in the uteri of rats subjected to activation of delayed implantation and experimentally induced decidualization. Treatment with 17β-estradiol or both 17β-estradiol and progesterone significantly diminished miR199a levels. Gain of function of miR199a in endometrial stromal cells isolated from rat uteri inhibited cell proliferation and promoted cell apoptosis. Loss of function of miR199a displayed opposite roles on cell proliferation and apoptosis. Further investigation uncovered a significant inverse association between the expression of miR199a and growth factor receptor-bound protein 10 (Grb10), an imprinted gene, and miR199a could bind to the 3′UTR of Grb10 to inhibit Grb10 translation. In addition, in vivo analysis found that the immunostaining of GRB10 was attenuated in the stroma or decidua from g.d.4 to 6, contrary to the enhancement of miR199a. Collectively, upregulation of miR199a in rat uterus during the receptive phase is regulated by blastocyst activation and uterine decidualization. Enforced miR199a expression suppresses cell proliferation partially through targeting Grb10.

scholarly journals Progesterone supplementation extends uterine receptivity for blastocyst implantation in mice

Reproduction ◽  
2007 ◽  
Vol 133 (2) ◽  
pp. 487-493 ◽  
Author(s):  
Haengseok Song ◽  
Kyuyong Han ◽  
Hyunjung Lim
Keyword(s):  
Cell Proliferation ◽  
Embryo Transfer ◽  
Thymidine Incorporation ◽  
Expression Patterns ◽  
Physiological Changes ◽  
Uterine Receptivity ◽  
Blastocyst Implantation ◽  
Progesterone Supplementation

We previously showed that blastocyst can initiate implantation beyond the normal ‘window’ of uterine receptivity on day 5 of pregnancy and pseudopregnancy (PSP) in mice. In this study, we investigated whether uterine receptivity for blastocyst implantation can be further extended on day 6 of PSP and the role of progesterone (P4) on this event. Embryo transfers, experimentally induced decidualization,in situhybridization and [3H]thymidine incorporation were performed. Blastocysts initiate attachment reaction within 48 h when transferred on day 5, but not on day 6 of PSP. Likewise, decidualization reaction occurred on days 4 and 5 of PSP, but completely failed on day 6. However, P4supplementation partially retains uterine receptivity for blastocyst implantation and decidualization on day 6 of PSP. In addition, certain indicators of uterine receptivity, such as cell proliferation profile and expression patterns of implantation-related genes were similarly observed on days 4 and 5 of PSP, but not on day 6. Consistent with embryo transfer and decidualization, exogenous administration of P4partially restores these indicators on day 6 of PSP. We concluded that critical physiological changes occur between days 4 and 5 of PSP, leading to uterine non-receptivity on day 6, but P4is able to extend the uterine receptivity through day 6.

scholarly journals A proteomic atlas of ligand-receptor interactions at the ovine maternal-fetal interface reveals the role of histone lactylation in uterine remodeling

2021 ◽  
Author(s):  
Qianying Yang ◽  
Juan Liu ◽  
Yue Wang ◽  
Wei Zhao ◽  
Wenjing Wang ◽  
...  
Keyword(s):  
Embryo Implantation ◽  
Redox Homeostasis ◽  
Integrated Analysis ◽  
Physical Interaction ◽  
Uterine Receptivity ◽  
In Vivo Models ◽  
Receptor Complexes ◽  
Maternal Fetal Interface

Abstract Well-orchestrated maternal-fetal crosstalk occurs via secreted ligands, interacting receptors, and coupled intracellular pathways between the conceptus and endometrium, and is essential for successful embryo implantation. However, previous studies mostly focus on either the conceptus or the endometrium in isolation. The lack of integrated analysis impedes our understanding of early maternal-fetal crosstalk. Herein, focusing on ligand–receptor complexes and coupled pathways at the maternal-fetal interface in sheep, we provide the first comprehensive proteomic map of ligand-receptor pathway cascades essential for embryo implantation. We demonstrate that these cascades are associated with cell adhesion and invasion, redox homeostasis, and the immune response. Candidate interactions and their physiological roles were further validated by functional experiments. We reveal the physical interaction of albumin and claudin 4 and their roles in facilitating embryo attachment to endometrium. We also demonstrate a novel function of enhanced conceptus glycolysis in remodeling uterine receptivity by inducing endometrial histone lactylation, a newly identified histone modification. Results from in vitro and in vivo models supported the essential role of lactate in inducing endometrial H3K18 lactylation and in regulating redox homeostasis and apoptotic balance to ensure successful implantation. By reconstructing a map of potential ligand-receptor pathway cascades at the maternal-fetal interface, our study presents new concepts for understanding molecular and cellular mechanisms that fine-tune conceptus-endometrium crosstalk during implantation. This provides more direct and accurate insights for developing potential clinical intervention strategies to improve pregnancy outcomes following both natural and assisted conception.

Uterine Epithelial Progesterone Receptor Governs Uterine Receptivity Through Epithelial Cell Differentiation

Endocrinology ◽  
2020 ◽  
Vol 161 (12) ◽  
Author(s):  
Mona Gebril ◽  
Yasushi Hirota ◽  
Shizu Aikawa ◽  
Yamato Fukui ◽  
Tetsuaki Kaku ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Epithelial Cell ◽  
Progesterone Receptor ◽  
Embryo Implantation ◽  
Rna Seq ◽  
Uterine Receptivity ◽  
Epithelial Cell Differentiation ◽  
Matrix Cell ◽  
Laser Capture ◽  
Embryo Attachment

Abstract Progesterone receptor (PGR) is indispensable for pregnancy in mammals. Uterine PGR responds to the heightened levels of ovarian progesterone (P4) after ovulation and regulates uterine gene transcription for successful embryo implantation. Although epithelial and stromal P4-PGR signaling may interact with each other to form appropriate endometrial milieu for uterine receptivity and the subsequent embryo attachment, it remains unclear what the specific roles of epithelial P4-PGR signaling in the adult uterus are. Here we generated mice with epithelial deletion of Pgr in the adult uterus (Pgrfl/flLtfCre/+ mice) by crossing Pgr-floxed and Ltf-Cre mice. Pgrfl/flLtfCre/+ mice are infertile due to the impairment of embryo attachment. Pgrfl/flLtfCre/+ uteri did not exhibit epithelial growth arrest, suggesting compromised uterine receptivity. Both epithelial and stromal expressions of P4-responsive genes decreased in Pgrfl/flLtfCre/+ mice during the peri-implantation period, indicating that epithelial Pgr deletion affects not only epithelial but stromal P4 responsiveness. In addition, uterine LIF, an inducer of embryo attachment, was decreased in Pgrfl/flLtfCre/+ mice. The RNA-seq analysis using luminal epithelial specimens dissected out by laser capture microdissection revealed that the signaling pathways related to extracellular matrix, cell adhesion, and cell proliferation are altered in Pgr fl/flLtf Cre/+ mice. These findings suggest that epithelial PGR controls both epithelial and stromal P4 responsiveness and epithelial cell differentiation, which provides normal uterine receptivity and subsequent embryo attachment.

scholarly journals Regulation of ARHGAP19 in the endometrial epithelium: a possible role in the establishment of uterine receptivity

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Jingjie Liang ◽  
Kui Li ◽  
Kaiyu Chen ◽  
Junyong Liang ◽  
Ti Qin ◽  
...  
Keyword(s):  
Rho Gtpase ◽  
Embryo Implantation ◽  
Luciferase Assay ◽  
Uterine Receptivity ◽  
Morphological Transformation ◽  
Upstream Regulator ◽  
Junctional Proteins

Abstract Background The establishment of uterine receptivity is essential for embryo implantation initiation and involves a significant morphological transformation in the endometrial epithelial cells (EECs). The remodeling of junctional complexes and membrane-associated cytoskeleton is crucial for epithelial transformation. However, little is known about how this process is regulated in EECs during the receptive phase. ARHGAP19 is a Rho GTPase-activating protein that participates in various cytoskeletal-related events, including epithelial morphogenesis. Here, we investigated the role of ARHGAP19 in endometrial epithelial transformation during the establishment of uterine receptivity. The upstream regulator of ARHGAP19 was also investigated. Methods ARHGAP19 expression was examined in mouse uteri during early pregnancy and in human EEC lines. The role of ARHGAP19 was investigated by manipulating its expression in EECs. The effect of ARHGAP19 on junctional proteins in EECs was examined by western blotting and immunofluorescence. The effect of ARHGAP19 on microvilli was examined by scanning electron microscopy. The upstream microRNA (miRNA) was predicted using online databases and validated by the dual-luciferase assay. The in vivo and in vitro effect of miRNA on endogenous ARHGAP19 was examined by uterine injection of miRNA agomirs and transfection of miRNA mimics or inhibitors. Results ARHGAP19 was upregulated in the receptive mouse uteri and human EECs. Overexpression of ARHGAP19 in non-receptive EECs downregulated the expression of junctional proteins and resulted in their redistribution. Meanwhile, upregulating ARHGAP19 reorganized the cytoskeletal structure of EECs, leading to a decline of microvilli and changes in cell configuration. These changes weakened epithelial cell polarity and promoted the transition of non-receptive EECs to a receptive phenotype. Besides, miR-192-5p, a miRNA that plays a key role in maintaining epithelial properties, was validated as an upstream regulator of ARHGAP19. Conclusion These results suggested that ARHGAP19 may contribute to the transition of EECs from a non-receptive to a receptive state by regulating the remodeling of junctional proteins and membrane-associated cytoskeleton.

scholarly journals AMPK is required for uterine receptivity and normal responses to steroid hormones

Reproduction ◽  
2020 ◽  
Vol 159 (6) ◽  
pp. 707-717 ◽  
Author(s):  
Richard M Griffiths ◽  
Cindy A Pru ◽  
Susanta K Behura ◽  
Andrea R Cronrath ◽  
Melissa L McCallum ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Epithelial Cell ◽  
Ovarian Function ◽  
Marker Genes ◽  
Epithelial Cell Proliferation ◽  
Uterine Receptivity ◽  
Serum Progesterone ◽  
Aberrant Expression ◽  
Stromal Compartment ◽  
Female Mice

We previously demonstrated that 5′-AMP-activated protein kinase (AMPK) is essential for normal reproductive functions in female mice. Conditional ablation of Prkaa1 and Prkaa2, genes that encode the α1 and α2 catalytic domains of AMPK, resulted in early reproductive senescence, faulty artificial decidualization, uterine inflammation and fibrotic postparturient endometrial regeneration. We also noted a delay in the timing of embryo implantation in Prkaa1/2d/d female mice, suggesting a role for AMPK in establishing uterine receptivity. As outlined in new studies here, conditional uterine ablation of Prkaa1/2 led to an increase in ESR1 in the uteri of Prkaa1/2d/d mice, resulting in prolonged epithelial cell proliferation and retention of E2-induced gene expression (e.g. Msx1, Muc1, Ltf) through the implantation window. Within the stromal compartment, stromal cell proliferation was reduced by five-fold in Prkaa1/2d/d mice, and this was accompanied by a significant decrease in cell cycle regulatory genes and aberrant expression of decidualization marker genes such as Hand2, Bmp2, Fst and Inhbb. This phenotype is consistent with our prior study, demonstrating a failure of the Prkaa1/2d/d uterus to undergo decidualization. Despite these uterine defects, ovarian function seemed to be normal following ablation of Prkaa1/2 from peri-ovulatory follicles in which ovulation, luteinization and serum progesterone levels were not different on day 5 of pregnancy or pseudopregnancy between Prkaa1/2fl/fl and Prkaa1/2d/d mice. These cumulative findings demonstrate that AMPK activity plays a prominent role in mediating several steroid hormone-dependent events such as epithelial cell proliferation, uterine receptivity and decidualization as pregnancy is established.

scholarly journals Uterine receptivity, embryo attachment, and embryo invasion: Multistep processes in embryo implantation

2019 ◽  
Vol 18 (3) ◽  
pp. 234-240 ◽  
Author(s):  
Yamato Fukui ◽  
Yasushi Hirota ◽  
Mitsunori Matsuo ◽  
Mona Gebril ◽  
Shun Akaeda ◽  
...  
Keyword(s):  
Embryo Implantation ◽  
Uterine Receptivity ◽  
Embryo Attachment

scholarly journals Crucial Role of Reactive Oxygen Species (ROS) for the Proapoptotic Effects of Indirubin Derivatives in Cutaneous SCC Cells

Antioxidants ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 1514
Author(s):  
Jiaqi Zhu ◽  
Peter Langer ◽  
Claas Ulrich ◽  
Jürgen Eberle
Keyword(s):  
Reactive Oxygen Species ◽  
Cell Proliferation ◽  
Reactive Oxygen ◽  
Oxygen Species ◽  
Antitumor Effects ◽  
Apoptosis Protein ◽  
High Incidence ◽  
Induced Apoptosis ◽  
Transcription 3

Efficient drugs are needed for countering the worldwide high incidence of cutaneous squamous cell carcinoma (cSCC) and actinic keratosis. Indirubin derivatives represent promising candidates, but their effects in cSCC cells have not been reported before. Here, we investigated the efficacy of three indirubin derivatives (DKP-071, -073 and -184) in four cSCC cell lines. High efficacy was seen in SCL-I, SCL-II, SCC-12 and SCC-13, resulting in up to 80% loss of cell proliferation, 60% loss of cell viability and 30% induced apoptosis (10 µM). Apoptosis was further enhanced in combinations with TNF-related apoptosis-inducing ligand (TRAIL). Induction of reactive oxygen species (ROS) appeared as critical for these effects. Thus, antioxidative pretreatment completely abolished apoptosis as well as restored cell proliferation and viability. Concerning the pathways, complete activation of caspases cascades (caspases-3, -4, -6, -7, -8 and -9), loss of mitochondrial membrane potential, activation of proapoptotic PKCδ (protein kinase C delta), inhibition of STAT3 (signal transducer and activator of transcription 3), downregulation of antiapoptotic XIAP (X-linked inhibitor of apoptosis protein) and survivin as well as upregulation of the proapoptotic Bcl-2 protein Puma and the cell cycle inhibitor p21 were obtained. Importantly, all activation steps were prevented by antioxidants, thus proving ROS as a master regulator of indirubins’ antitumor effects. ROS induction presently develops as an important issue in anticancer therapy.

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