Uterine Compartments Research Articles

Endometriosis affects natural and ART fertility in different ways: let's look at the whole patient and not at the single lesion

When considering the typical lesions associated with endometriosis, such as endometriomas, and pelvic adherences involving the tubes, it is very clear how this pathology may impair both natural and assisted reproductive technology (ART) fertility. It may be more difficult for clinicians to recognize that endometriosis can reduce female fertility potential through other mechanisms which may be independent of direct damage to ovarian reserve and tubal function. The most recent clinical studies have shown that endometriosis is associated with increased risk of infertility, independent of the type of endometriosis (ovarian, peritoneal and deep endometriosis). In the IVF setting, the cumulative live birth rate in women with endometriosis has been reported to be significantly lower compared with women without endometriosis. Endometriosis is a complex, multifactorial condition that encompasses not only the presence of endometriotic lesions, but also involves women's sexuality, uterine and ovarian compartment. Endometriosis should always be considered a severe risk factor for infertility and ART failure.

Membrane-bound progesterone receptors in the canine uterus and placenta; possible targets in the maintenance of pregnancy

To date, the biological functions of P4 within the canine placenta have been attributed to maternal stroma-derived decidual cells as the only placental cells expressing the nuclear P4 receptor (PGR). However, P4 can also exert its effects via membrane-bound receptors. To test the hypothesis that membrane-bound P4 receptors are involved in regulating placental function in the dog, the expression of mPRα, -β, -γ, PGRMC1 and -2 was investigated in the uterine and placental compartments derived from different stages of pregnancy and from prepartum luteolysis. Further, to assess the PGR signaling-mediated effects upon membrane P4 receptors in canine decidual cells, in vitro decidualized dog uterine stromal (DUS) cells were treated with type II antigestagens (aglepristone or mifepristone). The expression of all membrane P4 receptors was detectable in reproductive tissues and in DUS cells. The main findings indicate their distinguishable placental spatio-temporal distribution; PGRMC2 was predominantly found in decidual cells, PGRMC1 was strong in maternal endothelial compartments, and syncytiotrophoblast showed abundant levels of mPRα and mPRβ. In vitro decidualization was associated with increased expression of PGRMC1 and -2, while their protein levels were diminished by antigestagen treatment. The involvement of membrane-bound P4 signaling in the regulation of canine placental function is implied, with P4 effects being directly exerted through maternal and fetal cellular compartments. The indirect effects of PGR might involve the modulation of membrane-bound receptors availability in decidual cells, implying a self-regulatory loop of P4 in regulating the availability of its own receptors in the canine placenta.

A post-mortem analysis of tenofovir, lamivudine, efavirenz and fluconazole penetration in female genital tissues.

Optimal penetration of anti-infectives in the female genital tract (FGT) is paramount in the treatment and prevention of infectious diseases. While exposure of anti-infectives in lower FGT tissues (e.g. cervix, vagina) has been described, little data exist on upper genital tissues (e.g. ovary, uterus). Autopsies were performed and post-mortem tissues were collected within 24 h of death for female participants with advanced HIV in Uganda (n = 27). Tenofovir, lamivudine, efavirenz and fluconazole concentrations were measured using LC-MS/MS in plasma, ovarian, uterine, cervical and vaginal tissues. Tissue penetration was calculated as tissue-to-plasma concentration ratios (TPRs). TPRs of tenofovir, lamivudine and fluconazole were highest in vaginal tissue (medians 1.86, 1.83 and 0.94, respectively), while the TPR of efavirenz was highest in ovarian tissue (median 0.65). With cervix as a reference compartment, vaginal TPRs were significantly higher than cervical for all four drugs; TPRs of efavirenz in uterine and ovarian compartments were also significantly higher than cervical. Most of the post-mortem FGT samples had a TPR of greater than 1 for tenofovir and lamivudine, while less than 50% had a TPR of greater than 1 for both efavirenz and fluconazole. Penetration of anti-infectives was not homogeneous among the FGT compartments. Approximately 70% of FGT tissues had a TPR of greater than 1 for tenofovir and lamivudine, favouring the prevention of local HIV replication and transmission in the FGT.

Fertility-sparing surgery for diffuse adenomyosis: a narrated, stepwise approach to the Osada procedure

To equip reproductive surgeons with an approach to the Osada procedure and critical prophylactic hemostatic measures that optimize perioperative outcomes. Stepwise demonstration of the Osada procedure with narrated video footage. Definitive management of symptomatic adenomyosis requires hysterectomy. However, adenomyomectomy can improve symptoms and restore anatomy while maintaining fertility potential. Limited but comparable perioperative outcomes exist for minimally invasive methods of adenomyomectomy, and most involve resection of focal, not diffuse, adenomyosis. Among the literature involving resection of diffuse adenomyosis using minimally invasive methods, relatively small volumes of resected tissue are reported and none include obstetric outcomes. Most published reports for excision of diffuse adenomyosis involve laparotomic resection, likely because of specific intraoperative challenges curtailed by this approach. In response, a laparoscopic-assisted laparotomic approach was developed in 2011 by Dr. Hisao Osada, a reproductive surgeon in Japan. This procedure involves aggressive excision of adenomyotic tissue with prophylactic hemostatic techniques and subsequent uterine wall reconstruction using a triple-flap method. Compared with other excisional methods for diffuse adenomyomectomy, the Osada procedure has the best reported obstetric outcomes. A 37-year-old nulliparous female presented with pelvic pain, bulk symptoms, abnormal uterine bleeding, and infertility. Physical examination demonstrated a 20-week, bulky uterus with limited bimanual mobility. Her endometrial cavity was inaccessible because of marked anatomic distortion. Magnetic resonance imaging revealed marked abnormality of her endometrial contour because of a 15 cm adenomyoma with diffuse adenomyomatous tissue in the posterior uterine compartment. Prior interventions included a trial of combined hormonal contraceptive, leuprolide acetate, and tranexamic acid. She was interested in fertility-sparing adenomyomectomy to address symptoms and fertility potential and chose to proceed with the Osada procedure. She was optimized medically with oral and parenteral iron therapy to bring her hemoglobin from 55-111 g/L preoperatively. Institutional review board approval and informed consent from the patient were obtained. The Osada procedure was performed using the following 8 surgical steps: Systemic administration of tranexamic acid was also administered intraoperatively. Perioperative blood loss, anatomic normalization, symptom remediation, and maintenance of fertility potential. Perioperative blood loss was minimal, 469 g of adenomyotic tissue was extracted, and discharge was on postoperative day 2 without any complications. Three months later, cyclic pain and bleeding had improved markedly, ultrasound confirmed Doppler flow throughout the uterus, hysterosalpingogram demonstrated a nonobliterated endometrial cavity and tubal patency, and magnetic resonance imaging confirmed normalized uterine dimensions measuring 11 × 7 cm from 19 × 10 cm. Most literature supports waiting at least 6-12 months and until demonstration of normalized uterine blood flow in the operated area before attempting conception. Fertility-sparing excision of diffuse adenomyosis can be achieved safely using the Osada procedure, following the 8 discrete steps demonstrated in this video. Reproductive surgeons can reference this video to teach and maintain this important procedure.

P38α MAPK is a gatekeeper of uterine progesterone responsiveness at peri-implantation via Ube3c-mediated PGR degradation

Estrogen and progesterone specify the establishment of uterine receptivity mainly through their respective nuclear receptors, ER and PR. PR is transcriptionally induced by estrogen-ER signaling in the endometrium, but how the protein homeostasis of PR in the endometrium is regulated remains elusive. Here, we demonstrated that the uterine-selective depletion of P38α derails normal uterine receptivity ascribed to the dramatic down-regulation of PR protein and disordered progesterone responsiveness in the uterine stromal compartment, leading to defective implantation and female infertility. Specifically, Ube3c, an HECT family E3 ubiquitin ligase, targets PR for polyubiquitination and thus proteasome degradation in the absence of P38α. Moreover, we discovered that P38α restrains the polyubiquitination activity of Ube3c toward PR by phosphorylating the Ube3c at serine741 . In summary, we provided genetic evidence for the regulation of PR protein stability in the endometrium by P38α and identified Ube3c, whose activity was modulated by P38α-mediated phosphorylation, as an E3 ubiquitin ligase for PR in the uterus.

Effects of supplemental flaxseed on the ovarian and uterine functions of adult cycling mice.

Dietary fatty acids (FA) have an effect on animal reproduction. The purpose of the study was to find out how supplemental flaxseed can modulate the FA metabolism and how FA can influence the release of ovarian hormones and functions of the uteri through the modulation of their specific receptors. Cycling mice were fed a basal diet (control) and basal diet fortified with 10% flaxseed for 6 weeks to examine its influence on the structure and function of the ovaries and uteri, and serum levels of FA. Unlike controls (30%), 100% of mice fed flaxseed exposed oestrus at the end of the supplemental period. Serum FA were analysed using gas chromatography. The ovaries and uteri underwent histological and immunohistochemical analyses, respectively. The ovarian fragments were cultured with or without follicle-stimulating hormone and culture media were analysed for progesterone (P4) and oestradiol-17β (E2) using immunoassays. Dietary flaxseed increased the serum FA concentration, sizes of the ovaries and primary follicles, the release of P4 and E2, the thickness of endometrium and myometrium, and altered the expression of oestrogen and progesterone receptors in all uterine compartments. Dietary flaxseed can promote ovarian steroidogenesis and uterine activity in the mouse.

Spatial transcriptomics analysis of uterine gene expression in enhancer of zeste homolog 2 conditional knockout mice†.

Histone proteins undergo various modifications that alter chromatin structure, including addition of methyl groups. Enhancer of zeste homolog 2 (EZH2) is a histone methyltransferase that methylates lysine residue 27, and thereby suppresses gene expression. EZH2 plays integral roles in the uterus and other reproductive organs. We have previously shown that conditional deletion of uterine EZH2 results in increased proliferation of luminal and glandular epithelial cells, and RNA-seq analyses reveal several uterine transcriptomic changes in Ezh2 conditional (c) knockout (KO) mice that can affect estrogen signaling pathways. To pinpoint the origin of such gene expression changes, we used the recently developed spatial transcriptomics (ST) method with the hypotheses that Ezh2cKO mice would predominantly demonstrate changes in epithelial cells and/or ablation of this gene would disrupt normal epithelial/stromal gene expression patterns. Uteri were collected from ovariectomized adult WT and Ezh2cKO mice and analyzed by ST. Asb4, Cxcl14, Dio2, and Igfbp5 were increased, Sult1d1, Mt3, and Lcn2 were reduced in Ezh2cKO uterine epithelium vs. WT epithelium. For Ezh2cKO uterine stroma, differentially expressed key hub genes included Cald1, Fbln1, Myh11, Acta2, and Tagln. Conditional loss of uterine Ezh2 also appears to shift the balance of gene expression profiles in epithelial vs. stromal tissue toward uterine epithelial cell and gland development and proliferation, consistent with uterine gland hyperplasia in these mice. Current findings provide further insight into how EZH2 may selectively affect uterine epithelial and stromal compartments. Additionally, these transcriptome data might provide mechanistic understanding and valuable biomarkers for human endometrial disorders with epigenetic underpinnings.

P.154 Anaesthetic management of a dichorionic pregnancy in separate uterine compartments

Introduction: Bicornuate uterus is a rare anomaly of the Mullerian duct, estimated to occur in 1 in 250 women. [1] Slavchev S, Kostov S, Yordanov A. Pregnancy and childbirth in uterus didelphys. Medicina (Kaunas).2020; 56 198. Google Scholar Women with a bicornuate uterus are at greater risk of recurrent miscarriages, preterm birth, malpresentation and disruption to fetal growth. We present a case of a woman with a bicornuate uterus with a twin in each cavity who delivered by caesarean section using two separate uterine incisions. We discuss preoperative anaesthetic considerations, our anaesthetic technique and postoperative care.

Early-Pregnancy Dydrogesterone Supplementation Mimicking Luteal-Phase Support in ART Patients Did Not Provoke Major Reproductive Disorders in Pregnant Mice and Their Progeny

Progestogens are frequently administered during early pregnancy to patients undergoing assisted reproductive techniques (ART) to overcome progesterone deficits following ART procedures. Orally administered dydrogesterone (DG) shows equal efficacy to other progestogens with a higher level of patient compliance. However, potential harmful effects of DG on critical pregnancy processes and on the health of the progeny are not yet completely ruled out. We treated pregnant mice with DG in the mode, duration, and doses comparable to ART patients. Subsequently, we studied DG effects on embryo implantation, placental and fetal growth, fetal-maternal circulation, fetal survival, and the uterine immune status. After birth of in utero DG-exposed progeny, we assessed their sex ratios, weight gain, and reproductive performance. Early-pregnancy DG administration did not interfere with placental and fetal development, fetal-maternal circulation, or fetal survival, and provoked only minor changes in the uterine immune compartment. DG-exposed offspring grew normally, were fertile, and showed no reproductive abnormalities with the exception of an altered spermiogram in male progeny. Notably, DG shifted the sex ratio in favor of female progeny. Even though our data may be reassuring for the use of DG in ART patients, the detrimental effects on spermatogenesis in mice warrants further investigations and may be a reason for caution for routine DG supplementation in early pregnancy.

In the mouse, prostaglandin D2 signalling protects the endometrium against adenomyosis.

Adenomyosis is characterised by epithelial gland and mesenchymal stroma invasion of the uterine myometrium. Adenomyosis is an oestrogen-dependent gynaecological disease in which a number of factors, such as inflammatory molecules, prostaglandins (PGs), angiogenic factors, cell proliferation and extracellular matrix remodelling proteins, also play a role as key disease mediators. In this study, we used mice lacking both lipocalin and hematopoietic-PG D synthase (L- and H-Pgds) genes in which PGD2 is not produced to elucidate PGD2 roles in the uterus. Gene expression studied by real-time PCR and hormone dosages performed by ELISA or liquid chromatography tandem mass spectroscopy in mouse uterus samples showed that components of the PGD2 signalling pathway, both PGDS and PGD2-receptors, are expressed in the mouse endometrium throughout the oestrus cycle with some differences among uterine compartments. We showed that PGE2 production and the steroidogenic pathway are dysregulated in the absence of PGD2. Histological analysis of L/H-Pgds-/- uteri, and immunohistochemistry and immunofluorescence analyses of proliferation (Ki67), endothelial cell (CD31), epithelial cell (pan-cytokeratin), myofibroblast (α-SMA) and mesenchymal cell (vimentin) markers, identify that 6-month-old L/H-Pgds-/- animals developed adenomyotic lesions, and that disease severity increased with age. In conclusion, this study suggests that the PGD2 pathway has major roles in the uterus by protecting the endometrium against adenomyosis development. Additional experiments, using for instance transcriptomic approaches, are necessary to fully determine the molecular mechanisms that lead to adenomyosis in L/H-Pgds-/- mice and to confirm whether this strain is an appropriate model for studying the human disease.

Abstract PO041: Everolimus and MK-2206 reverse P4-resistant endometrial hyperplasia in Mig-6 mutant mice

Abstract Endometrial cancer is the most frequent type of gynecological tumor in the United States. Hysterectomy, the surgical removal of the uterus, is the main therapy for endometrial cancer, but this is not an option for women seeking to preserve their fertility. In cases of early-stage endometrial cancer and endometrial hyperplasia, hormonal therapy is a vital nonsurgical option. However, progesterone (P4) resistance robs women of the option to choose hormone therapy to preserve their uterus. Therefore, developing nonsurgical treatments for endometrial cancer remains an essential goal for women who wish to maintain endometrial functions and women in high-risk surgical conditions. Mitogen-inducible gene 6 (MIG-6) acts as a tumor suppressor during endometrial tumorigenesis and as a P4 target gene in humans and mice. We developed mice with the conditional knockout of Mig-6 in all uterine compartments (Pgrcre/+Mig-6f/f; Mig-6KO) and endometrial epithelial cell-specific Mig-6 knockout (Sprr2fcre/+Mig-6f/f; Mig-6Ep-KO) mice. P4 treatment abated the development of endometrial hyperplasia and restored morphological and histological characteristics of the uterus of Mig-6Ep-KO mice, but Mig-6Ep-KO mice showed P4 resistance. P4 treatment reduced cell proliferation which was accompanied by decreased AKT signaling. In order to investigate the inhibition effect of AKT/mTOR signaling as non-surgical treatment on P4-resistant endometrial hyperplasia, we treated Mig-6KO and Mig-6Ep-KO mice with either: 1) vehicle (PBS); 2) MK-2206 (360 mg/kg, once a week); or 3) everolimus (10mg/kg daily) at 10 weeks old (n=6/treatment/genotype) for 1 month. MK-2206 has completed phase II trials and inhibits AKT phosphorylation. Everolimus is FDA-approved mTOR inhibitor. The vehicle group of Mig-6KO and Mig-6Ep-KO mice manifested the endometrial hyperplasia phenotype. In contrast, IHC analysis showed that the groups receiving MK-2206 or everolimus experienced significantly decreased AKT phosphorylation and phosphorylated S6 levels, respectively, and decreased epithelial proliferation, resulting in restoration of normal uterine histology. These data suggest that everolimus and MK-2206 may be new non-surgical treatment options for P4-resistant endometrial hyperplasia. This information may help guide more effective non-surgical interventions in the future. The project described was supported by Grant Number P50CA098258 from the National Cancer Institute. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Cancer Institute or the National Institutes of Health. Citation Format: Tae Hoon Kim, Jae-Wook Jeong. Everolimus and MK-2206 reverse P4-resistant endometrial hyperplasia in Mig-6 mutant mice [abstract]. In: Proceedings of the AACR Virtual Special Conference: Endometrial Cancer: New Biology Driving Research and Treatment; 2020 Nov 9-10. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(3_Suppl):Abstract nr PO041.

The Cervicovaginal Mucus Barrier.

Preterm births are a global health priority that affects 15 million babies every year worldwide. There are no effective prognostic and therapeutic strategies relating to preterm delivery, but uterine infections appear to be a major cause. The vaginal epithelium is covered by the cervicovaginal mucus, which is essential to health because of its direct involvement in reproduction and functions as a selective barrier by sheltering the beneficial lactobacilli while helping to clear pathogens. During pregnancy, the cervical canal is sealed with a cervical mucus plug that prevents the vaginal flora from ascending toward the uterine compartment, which protects the fetus from pathogens. Abnormalities of the cervical mucus plug and bacterial vaginosis are associated with a higher risk of preterm delivery. This review addresses the current understanding of the cervicovaginal mucus and the cervical mucus plug and their interactions with the microbial communities in both the physiological state and bacterial vaginosis, with a focus on gel-forming mucins. We also review the current state of knowledge of gel-forming mucins contained in mouse cervicovaginal mucus and the mouse models used to study bacterial vaginosis.

Transforming growth factor beta signaling and decidual integrity in mice†.

Transforming growth factor beta (TGFβ) signaling regulates multifaceted reproductive processes. It has been shown that the type 1 receptor of TGFβ (TGFBR1) is indispensable for female reproductive tract development, implantation, placental development, and fertility. However, the role of TGFβ signaling in decidual development and function remains poorly defined. Our objective is to determine the impact of uterine-specific deletion of Tgfbr1 on decidual integrity, with a focus on the cellular and molecular properties of the decidua during development. Our results show that the developmental dynamics of the decidua is altered in TGFBR1 conditionally depleted uteri from embryonic day (E) 5.5 to E8.5, substantiated by downregulation of genes associated with inflammatory responses and uterine natural killer cell abundance, reduced presence of nondecidualized fibroblasts in the antimesometrial region, and altered decidual cell development. Notably, conditional ablation of TGFBR1 results in the formation of decidua containing more abundant alpha smooth muscle actin (ACTA2)-positive cells at the peripheral region of the antimesometrial side versus controls at E6.5-E8.5. This finding is corroborated by upregulation of a subset of smooth muscle marker genes in Tgfbr1 conditionally deleted decidua at E6.5 and E8.5. Moreover, increased cell proliferation and enhanced decidual ERK1/2 signaling were found in Tgfbr1 conditional knockout mice upon decidual regression. In summary, conditional ablation of TGFBR1 in the uterus profoundly impacts the cellular and molecular properties of the decidua. Our results suggest that TGFBR1 in uterine epithelial and stromal compartments is important for the integrity of the decidua, a transient but crucial structure that supports embryo development.

\u03b3\u03b4 T cells compose a developmentally regulated intrauterine population and protect against vaginal candidiasis

This most comprehensive analysis to date of γδ T cells in the murine uterus reveals them to compose a unique local T-cell compartment. Consistent with earlier reports, most cells expressed a canonical Vγ6Vδ1 TCR, and produced interleukin (IL)-17A upon stimulation. Nonetheless, contrasting with earlier reports, uterine γδ T cells were not obviously intraepithelial, being more akin to sub-epithelial Vγ6Vδ1+ T cells at several other anatomical sites. By contrast to other tissues however, the uterine compartment also included non-Vγ6+, IFN-γ-producing cells; was strikingly enriched in young mice; expressed genes hitherto associated with the uterus, including the progesterone receptor; and did not require microbes for development and/or maintenance. This notwithstanding, γδ T-cell deficiency severely impaired resistance to reproductive tract infection by Candida albicans, associated with decreased responses of IL-17-dependent neutrophils. These findings emphasise tissue-specific complexities of different mucosal γδ cell compartments, and their evident importance in lymphoid stress-surveillance against barrier infection.

TNF-Signaling Modulates Neutrophil-Mediated Immunity at the Feto-Maternal Interface During LPS-Induced Intrauterine Inflammation.

Accumulation of activated neutrophils at the feto-maternal interface is a defining hallmark of intrauterine inflammation (IUI) that might trigger an excessive immune response during pregnancy. Mechanisms responsible of this massive neutrophil recruitment are poorly investigated. We have previously showed that intraamniotic injection of LPS in rhesus macaques induced a neutrophil predominant inflammatory response similar to that seen in human IUI. Here, we demonstrate that anti-TNF antibody (Adalimumab) inhibited ~80% of genes induced by LPS involved in inflammatory signaling and innate immunity in chorio-decidua neutrophils. Consistent with the gene expression data, TNF-blockade decreased LPS-induced neutrophil accumulation and activation at the feto-maternal interface. We also observed a reduction in IL-6 and other pro-inflammatory cytokines but not prostaglandins concentrations in the amniotic fluid. Moreover, TNF-blockade decreased mRNA expression of inflammatory cytokines in the chorio-decidua but not in the uterus, suggesting that inhibition of TNF-signaling decreased the inflammation in a tissue-specific manner within the uterine compartment. Taken together, our results demonstrate a predominant role for TNF-signaling in modulating the neutrophilic infiltration at the feto-maternal interface during IUI and suggest that blockade of TNF-signaling could be considered as a therapeutic approach for IUI, the major leading cause of preterm birth.

Neonatal sepsis evaluation across the pond

One of the most uncommon, life-threatening and feared issues newborn clinicians struggle to measure is the likelihood that a newborn has acquired a bacterial infection during the process of birth....

2671 Lefort Colpocleisis

Video Objective To present the technical considerations and pearls to performing a LeFort Colpocleisis. Setting Patient is a 76 year old female with history of a vaginal bulge for the past 6 months and a failed pessary trial. She also reports urge-predominant mixed urinary incontinence with worsening stress urinary incontinence with reduction of the pelvic organ prolapse. She is not sexually active and does not plan on any future sexual activity. She does not have any abnormal pap smears and denies postmenopausal bleeding. Her past medical history is significant for multiple comorbid conditions including cardiac issues with a pacemaker, chronic kidney disease, hypertension and diabetes. Her past surgical history is complicated by aortic valve replacement (porcine valve), tubal ligation and a left total knee replacement. On exam she has a Stage IV uterine, anterior and posterior compartment prolapse with positive occult stress urinary incontinence. Urodynamic studies reveal stress urinary incontinence, no detrusor overactivity and an elevated post void residual (125 ml after 241 ml void). She was counseled regarding management options including conservative options (observation or repeat pessary trial), restorative procedures (vaginal vault suspension or sacrocolpopexy), or an obliterative procedure such as a LeFort Colpocleisis. Interventions The patient opted for a LeFort Colpocleisis. She underwent a Dilation and curettage, a LeFort colpocleisis, Posterior colpoperineorrhaphy, Cystoscopy and a Midurethral Sling Placement. Conclusion The technical considerations for performing a LeFort Colpocleisis include ruling out malignancy prior to surgery, adequate lateral channels for uterine drainage, the use of lidocaine with epinephrine, closure in multiple layers with excellent hemostasis and an aggressive posterior colpoperineorrhaphy.

A calcium-dependent phospholipase A2 (cPLA2) expression is regulated by MIG-6 during endometrial tumorigenesis

The ovarian steroid hormones, estrogen (E2) and progesterone (P4), are essential regulators of uterine biology. The imbalance of these ovarian steroid hormones leads to uterine diseases such as endometrial cancer, endometriosis, and infertility. Mitogen-inducible gene 6 (MIG-6) is an adaptor protein. MIG-6 mediates P4 signaling and acts as a tumor suppressor during endometrial tumorigenesis in both humans and mice. In previous studies, we developed the conditional knockout of Mig-6 in all uterine compartments (Pgrcre/+Mig-6f/f; Mig-6KO) and endometrial epithelial cell-specific Mig-6 knockout (Sprr2fcre/+Mig-6f/f; Mig-6Ep-KO) mice. Both mouse models developed endometrial hyperplasia and E2-dependent endometrial cancer. P4 treatment significantly decreases aberrant epithelial proliferation and AKT signaling in Mig-6Ep-KO mice but not in Mig-6KO mice. In the present study, we identified a calcium-dependent phospholipase A2 (cPla2) as one of the genes down-regulated by Mig-6 in the uterus. We performed immunohistochemistry and Western Blot analysis to investigate the regulation of cPLA2 by MIG-6 as well as determine the expression patterns of cPLA2 in the uterus. While the expression of cPLA2 was stronger at the uterine epithelial cells of Mig-6KO and Mig-6Ep-KO mice compared to control mice, P4 suppressed the expression of cPLA2 in Mig-6Ep-KO mice but not in Mig-6KO mice. To determine the ovarian steroid hormone regulation of cPLA2, we examined the expression of cPLA2 in ovariectomized control, Mig-6KO, Mig-6Ep-KO, and PRKO mice treated with P4 or E2. After P4 treatment, cPLA2 expression was remarkably reduced in Mig-6Ep-KO mice but not in Mig-6KO mice. However, the expression of cPLA2 was not changed in PRKO mice. Our results identified cPLA2 as a novel target of MIG-6 in the murine uterus and identified its important role during endometrial tumorigenesis.

Steroid concentrations in maternal serum and uterine histotroph in round stingrays (Urobatis halleri)

Despite a wide range of elasmobranch (sharks, skates and rays) matrotrophic strategies, and thus potentially diverse pathways for maternal-fetal hormone exchange, little attention has been given to uterine steroids during development. Round Stingrays (Urobatis halleri) with matrotrophic histotrophy were captured during every month of their annual reproductive season from post-ovulation to near parturition, and paired samples of plasma and histotroph were analyzed for a suite of steroid hormones using LC-ESI/MRM. Hormone concentrations within and between maternal and uterine compartments were compared using two markers of embryo development. Histotroph had consistently higher detection rates and concentrations of hormones than maternal plasma, especially during early pregnancy when embryos are yolk sac-dependent for nutrition. Peaks in histotroph testosterone concentrations preceded maternal plasma, suggesting that hormones were locally produced within the uterine compartment. Embryonic sexual differentiation based on the presence of visible claspers (male copulatory organs) coincided with peaks in histotroph progesterone, testosterone, 17-hydroxyprogesterone and estradiol, suggesting that, like mammalian pregnancy, elasmobranch embryonic steroids also contribute to their own developmental environment.

Follicle stimulating hormone receptor protein is expressed in ovine uterus during the estrous cycle and utero-placenta during early pregnancy: An immunohistochemical study

Follicle stimulating hormone (FSH) is a well characterized gonadotropin that controls primarily development and functions of ovarian follicles in mammalian species. FSH binds to a specific G protein-coupled receptor (FSHR) belonging to the glycoprotein hormone receptor family that plays an essential role in reproduction. Although the primary location of FSHR is in the gonads (mainly in ovarian follicles), FSHR protein and/or mRNA have also been detected in extragonadal female reproductive tissues including embryo, placenta, endometrium, cervix, ovarian cancer tissues, and/or endometriotic lesions in several species. To determine the pattern of FSHR expression in the uterus and placenta, uterine tissues were collected at the early, mid- and/or late luteal phases of the estrous cycle from non-treated or FSH-treated ewes, and utero-placental tissues were collected during early pregnancy followed by immunohistochemistry and image generation. FSHR was immunolocalized to several uterine and utero-placental compartments including luminal epithelium, endometrial glands and surrounding stroma, myometrium, and endothelium and vascular smooth muscle cells in endometrium, myometrium and mesometrium. Intensity of staining and distribution of FSHR in selected compartments differed and seems to depend on the stage of the estrous cycle or pregnancy, and FSH-treatment. These novel data demonstrate differential expression of FSHR protein indicating that FSH plays a specific role in regulation of uterine and utero-placenta functions in sheep.