uterine epithelial cells
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2021 ◽  
Vol 12 ◽  
Author(s):  
Mickey V. Patel ◽  
Daniel C. Hopkins ◽  
Fiona D. Barr ◽  
Charles R. Wira

Estradiol (E2) and progesterone (P) have potent effects on immune function in the human uterine endometrium which is essential for creating an environment conducive for successful reproduction. Type III/lambda (λ) interferons (IFN) are implicated in immune defense of the placenta against viral pathogens, which occurs against the backdrop of high E2 and P levels. However, the effect of E2 and P in modulating the expression and function of IFNλ1 in the non-pregnant human uterine endometrium is unknown. We generated purified in vitro cultures of human uterine epithelial cells and stromal fibroblast cells recovered from hysterectomy specimens. Poly (I:C), a viral dsRNA mimic, potently increased secretion of IFNλ1 by both epithelial cells and fibroblasts. The secretion of IFNλ1 by epithelial cells significantly increased with increasing age following poly (I:C) stimulation. Stimulation of either cell type with E2 (5x10-8M) or P (1x10-7M) had no effect on expression or secretion of IFNλ1 either alone or in the presence of poly (I:C). E2 suppressed the IFNλ1-induced upregulation of the antiviral IFN-stimulated genes (ISGs) MxA, OAS2 and ISG15 in epithelial cells, but not fibroblasts. Estrogen receptor alpha (ERα) blockade using Raloxifene indicated that E2 mediated its inhibitory effects on ISG expression via ERα. In contrast to E2, P potentiated the upregulation of ISG15 in response to IFNλ1 but had no effect on MxA and OAS2 in epithelial cells. Our results demonstrate that the effects of E2 and P on IFNλ1-induced ISGs are cell-type specific. E2-mediated suppression, and selective P-mediated stimulation, of IFNλ1-induced ISG expression in uterine epithelial cells suggest that the effects of IFNλ1 varies with menstrual cycle stage, pregnancy, and menopausal status. The suppressive effect of E2 could be a potential mechanism by which ascending pathogens from the lower reproductive tract can infect the pregnant and non-pregnant endometrium.


Author(s):  
Jessica S Dudley ◽  
Christopher R Murphy ◽  
Michael B Thompson ◽  
Bronwyn M McAllan

Abstract There are many different forms of nutrient provision in viviparous (live bearing) species. The formation of a placenta is one method where the placenta functions to transfer nutrients from mother to fetus (placentotrophy), transfer waste from the fetus to the mother and respiratory gas exchange. Despite having the same overarching function, there are different types of placentation within placentotrophic vertebrates, and many morphological changes occur in the uterus during pregnancy to facilitate formation of the placenta. These changes are regulated in complex ways but are controlled by similar hormonal mechanisms across species. This review describes current knowledge of the morphological and molecular changes to the uterine epithelium preceding implantation among mammals. Our aim is to identify the commonalities and constraints of these cellular changes to understand the evolution of placentation in mammals and propose directions for future research. We compare and discuss the complex modifications to the ultrastructure of uterine epithelial cells and show that there are similarities in the changes to the cytoskeleton and gross morphology of the uterine epithelial cells, especially of the apical and lateral plasma membrane of the cells during the formation of a placenta in all eutherians and marsupials studied to date. We conclude that further research is needed to understand the evolution of placentation among viviparous mammals, particularly concerning the level of placental invasiveness, hormonal control and genetic underpinnings of pregnancy in marsupial taxa.


2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Hyunji Byun ◽  
Sojung Kwon ◽  
Kay-Uwe Wagner ◽  
Hyejin Shin ◽  
Hyunjung Jade Lim

Abstract Background The tumor susceptibility gene 101 (Tsg101), a component of the endosomal sorting complex required for transport (ESCRT) complex I, is involved in multiple biological processes involving endomembranous structures and the plasma membrane. The role of Tsg101 in the uterine epithelium was investigated in Tsg101 floxed mice crossed with Lactoferrin-iCre mice (Tsg101d/d). Methods Tsg101d/d mice were bred with stud male mice and the status of pregnancy was examined on days 4 and 6. Histological analyses were performed to examine the uterine architecture. Immunofluorescence staining of several markers was examined by confocal microscopy. Uterine epithelial cells (UECs) were isolated from Tsg101f/f and Tsg101d/d mice, and the expression of necroptosis effectors was examined by RT-PCR, western blotting, and immunofluorescence staining. UECs were also subjected to RNA expression profiling. Results Tsg101d/d female mice were subfertile with implantation failure, showing unattached blastocysts on day 6 of pregnancy. Histological and marker analyses revealed that some Tsg101d/d day 4 pregnant uteri showed a disintegrated uterine epithelial structure. Tsg101d/d UECs began to degenerate within 18 h of culture. In UECs, expression of necroptosis effectors, such as RIPK1, RIPK3, and MLKL were first confirmed. UECs responded to a stimulus to activate necroptosis and showed increased cell death. Conclusions Tsg101 deficiency in the uterine epithelium causes implantation failure, which may be caused by epithelial defects. This study provides evidence that UECs harbor a necroptotic machinery that responds to death-inducing signals. Thus, Tsg101 expression in the uterine epithelium is required for normal pregnancy in mice.


Cells ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 1194
Author(s):  
Vanessa Mancini ◽  
Alexandra C. Schrimpe-Rutledge ◽  
Simona G. Codreanu ◽  
Stacy D. Sherrod ◽  
John A. McLean ◽  
...  

Here we report the use of a microfluidic system to assess the differential metabolomics of murine embryos cultured with endometrial cells-conditioned media (CM). Groups of 10, 1-cell murine B6C3F1×B6D2F1 embryos were cultured in the microfluidic device. To produce CM, mouse uterine epithelial cells were cultured in potassium simplex optimized medium (KSOM) for 24 h. Media samples were collected from devices after 5 days of culture with KSOM (control) and CM, analyzed by reverse phase liquid chromatography and untargeted positive ion mode mass spectrometry analysis. Blastocyst rates were significantly higher (p < 0.05) in CM (71.8%) compared to control media (54.6%). We observed significant upregulation of 341 compounds and downregulation of 214 compounds in spent media from CM devices when compared to control. Out of these, 353 compounds were identified showing a significant increased abundance of metabolites involved in key metabolic pathways (e.g., arginine, proline and pyrimidine metabolism) in the CM group, suggesting a beneficial effect of CM on embryo development. The metabolomic study carried out in a microfluidic environment confirms our hypothesis on the potential of uterine epithelial cells to enhance blastocyst development. Further investigations are required to highlight specific pathways involved in embryo development and implantation.


2021 ◽  
Author(s):  
Hyunji Byun ◽  
Sojung Kwon ◽  
Kay-Uwe Wagner ◽  
Hyejin Shin ◽  
Hyunjung Jade Lim

Abstract Background: The tumor susceptibility gene 101 (Tsg101), a component of the endosomal sorting complex required for transport (ESCRT) complex I, is involved in multiple biological processes involving endomembranous structures and the plasma membrane. The role of Tsg101 in the uterine epithelium was investigated in Tsg101 floxed mice crossed with Lactoferrin-iCre mice (Tsg101d/d).Methods: Tsg101d/d mice were bred with stud male mice and the status of pregnancy was examined on days 4 and 6. Histological analyses were performed to examine the uterine architecture. Immunofluorescence staining of several markers was examined by confocal microscopy. Uterine epithelial cells (UECs) were isolated from Tsg101f/f and Tsg101d/d mice, and the expression of necroptosis effectors was examined by RT-PCR, western blotting, and immunofluorescence staining. UECs were also subjected to RNA expression profiling.Results: Tsg101d/d female mice were subfertile with implantation failure, showing unattached blastocysts on day 6 of pregnancy. Histological and marker analyses revealed that some Tsg101d/d day 4 pregnant uteri showed a disintegrated uterine epithelial structure. Tsg101d/d UECs began to degenerate within 18 h of culture. In UECs, expression of necroptosis effectors, such as RIPK1, RIPK3, and MLKL were first confirmed. UECs responded to a stimulus to activate necroptosis and showed increased cell death. Conclusions: Tsg101 deficiency in the uterine epithelium causes implantation failure, which accompanies epithelial defects. This study provides evidence that UECs harbor a necroptotic machinery that responds to death-inducing signals. Thus, Tsg101 expression in the uterine epithelium is required for normal pregnancy in mice.


Author(s):  
Laura A. Lindsay ◽  
Reeja F. Nasir ◽  
Samson N. Dowland ◽  
Romanthi J. Madawala ◽  
Christopher R. Murphy

GYNECOLOGY ◽  
2020 ◽  
Vol 22 (6) ◽  
pp. 62-67
Author(s):  
Irina N. Vorobtsova ◽  
Natalya I. Tapilskaya ◽  
Elena S. Orlova ◽  
Nikolay N. Rukhlyada ◽  
Sergei N. Proshin ◽  
...  

Relevance. Human immunodeficiency virus (HIV) infection is an independent factor in reduced fertility and a risk factor for miscarriage. There are some date an endometrial receptivity of HIV-infected patients has changed that plays an important role in embryo invasion, but the true reasons for the decrease in fertility rate in HIV infection remain unknown. Aim. Study of the expression of CD20, CD56 and TLR9 antigens on uterine epithelial cells of HIV-infected patients and the effectiveness of treatment for chronic endometritis by sodium nucleospermate. Materials and methods. This parallel-group study was done at two centres in the Russia. Participants were adults women aged 26 to 49 years (mean age 33.352.9 years), who were HIV-infected (n=12) and HIV-negative (22). An immunocytochemical study of endometrial biopsies taken on the 710th day of the menstrual cycle before and after treatment was done. The course of treatment with sodium nucleospermate was 42 days. Results. The expression level of CD56 and TLR9 in HIV-infected patients was 7.640.92% and 0.330.18%, respectively, and significantly differed from the expression levels in HIV-seronegative patients. There was a decrease in the expression levels of CD20 and CD56 and an increase in the expression levels of TLR9 in all groups of patients after treatment with sodium nucleospermate. Conclusion. A decrease TLR9 expression on uterine epithelial cells in HIV-infected patients showing lack of ability of innate immunity to eliminate pathogens associated with subclinical inflammation and it correlates with an increase in the expression of markers of chronic endometritis.


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