Progesterone Receptor Gene Research Articles

Unconservative_15_2570409 suppresses progesterone receptor expression in the granulosa cells of Hu sheep

Female fertility potential depends on the number of mature follicles; however, the underlying molecular mechanisms remain unclear. Based on previously generated miRNA and mRNA sequencing data of healthy ovarian follicles (>5 mm in diameter) isolated from Hu sheep with different prolificacy, we investigated the roles of a novel miRNA (unconservative_15_2570409) and the progesterone receptor (PGR) gene in follicular development. During the periovulatory phase, the expression of unconservative_15_2570409 and PGR was lower and higher, respectively, in the >5 mm follicles of high prolificacy (HP) ewes than in those of low prolificacy (LP) ewes and in the >3 mm follicles than in the smaller follicles of the HP ewes. Subsequently, the granulosa cells (GCs) of Hu sheep were used as an in vitro model. PGR overexpression significantly increased the mRNA expression of steroidogenic acute regulatory protein (StAR), 3-beta-hydroxysteroid dehydrogenase/isomerase (3β-HSD), and cytochrome P450 family 19 subfamily A member 1 (CYP19A1), which promoted the secretion of progesterone (P4) and estradiol (E2). PGR knockdown significantly downregulated the levels of StAR and 3β-HSD mRNA and decreased the production of P4, whereas no effects on CYP19A1 mRNA expression and E2 levels were observed. We also found the negative regulatory effect of unconservative_15_2570409 on the mRNA and protein expression of PGR by targeting the 3ʹ-untranslated region. The regulation of PGR levels resulted in a corresponding change in the ADAMTS1, PPAR-γ, and CTSL gene transcripts, which are important for follicular maturation and ovulation. Additionally, PGR, ADAMTS1, and PPAR-γ were predominantly localized in the GCs. Collectively, our results suggest that by regulating PGR expression and consequently affecting the expression of target genes and steroidogenesis, unconservative_15_2570409 plays a role in follicular development during the periovulatory stage, which provides novel insights into the molecular mechanisms affecting Hu sheep prolificacy.

Abstract 1679: Characterization and establishment of triple negative breast cancer patient derived xenograft derived from a Nigerian woman

Abstract Extensive literature has demonstrated the use of fresh human breast tumour tissue surgically implanted in immunodeficient mice to establish patient-derived xenograft (PDX) as a therapeutic assessment tool. Unfortunately, there is a paucity of genomic research using PDX as a preclinical tool for breast cancer metastasis in Nigerian women. Previous studies of rodent models have concentrated on cell line xenograft of other tumours usually of Caucasian or African American counterpart. To overcome this limitation, Triple-negative breast cancer (TNBC) was derived from a consenting 32-year-old Nigerian woman who had neoadjuvant therapy and had undergone a mastectomy. Triple-negative breast cancer lacks expression of hormone receptors including human epidermal receptor (HER2-), estrogen receptor (ER-) and progesterone receptor (PR-) genes. TNBC xenograft was established by orthotopic surgical implantation of breast cancer tissue fragments of 2mm3 in size with cultrex in the cleared breast pads of immunosuppressed mice and a further two passages. The size of the implanted tumour was checked twice weekly. Haematology & eosin histology and immunohistochemistry analysis were used to structurally and functionally characterize the tumour xenograft and primary tumour respectively. Real-time quantitative polymerase chain reaction (RQ-PCR) was used to evaluate the genomic correlation between primary tumour and tumour xenograft. Target genes compared between the tumour xenograft and primary tumours included CXCL16; CLD10; CDC42EP3; CXCL16; EPSTI1; PALMD and ZAN. Reference genes RLP13A and GAPDH were selected as assays for endogenous controls. All tumour tissues were mechanically homogenized and lysed in guanidine isothiocyanate buffer from which RNA was extracted and converted to cDNA by random hexamer priming. RQ-PCR was performed using TaqMan chemistries. This was validated by DNA sequencing of the xenografts tumour and primary tumour using Nanopore DNA sequencing, and Ligation Sequencing ID2 chemistry. The DNA sequences were aligned with Meg Align Pro The xenografts appear to maintain characteristic human histopathology, tumour-marker production. None of the tumours showed hormone sensitivity. There was a local invasion of breast cancer in the bone. The pairwise alignment yielded significant percentage relativity. Citation Format: Uzoamaka Adaobi Okoli, Michael Tochukwu Okafor, Kenneth A. Agu, Samuel R. Ohayi, Emmanuel O. Nna, Chioli P. Chijioke, Iroka J. Udeniya. Characterization and establishment of triple negative breast cancer patient derived xenograft derived from a Nigerian woman [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1679.

Evaluation of progesterone and progesterone receptor gene PROGINS polymorphism in the development of some forms of premenstrual syndrome

One of the most common neuroendocrine syndromes in gynecology is premenstrual syndrome (PMS). The article is devoted to study of pro-gesterone level and frequency of polymorphic variants T1T2 of progesterone receptor gene PROGINS, as well as their possible relationship in patients with various forms of PMS. The object of the study were 50 women of reproductive age with diagnosis of PMS, including 25 women with edematous form of the disease, 25 – with neuropsychical form. Mild form of disease had 25 women, severe form – also 25 persons. Control group consisted of 25 women without diagnosis of PMS. Progesterone level was determined in blood serum twice in the first and second phase of menstrual cycle. Polymorphism T1T2 of progesterone gene receptor PROGINS was studied using polymerase chain reaction.We found hypoprogesteronemia of luteal phase of menstrual cycle in women with PMS, especially pronounced in persons with edematous and severe forms of the disease (p<0.05).Polymorphic variant T1T2 of gene PROGINS can be considered as a marker of development of PMS. T1T1 genotype was significantly as-sociated with development of edematous form (χ2=4.50; р=0.03) and its presence may indicate a tendency to develop of severe form of PMS (χ2=3.21; р=0.07). T2 allele can be considered as a protective mechanism for the appearance of PMS, particularly its edematous and severe forms, in which the presence of T2 allele reduces the risk of PMS, respectively 3.0 times (χ2=4.50; p=0.03, OR=0.20 95% CI 0.05–0.78; p=0.02) and 1,71 times (χ2=3.21, p=0.07, OR=0.27, 95% CI 0.08–0.95, p=0.04) compared with control group. Reduced blood level of progesterone in women with PMS correlated with the presence of T1T1 genotype of gene PROGINS. By edematous, neuro-psychical, severe and mild forms of PMS and the presence of T1T1 genotype there were significantly low levels of this hormone in the second phase of menstrual cycle (p<0.05) relative to women with the same genotype in the control group of. Furthermore, there was no statistically significant difference in progesterone level in women with T1T2 genotype in these groups compared with healthy women with same genotype (p>0.05)

Perinatal exposure to glyphosate or a glyphosate-based formulation disrupts hormonal and uterine milieu during the receptive state in rats

We investigated the effects of perinatal exposure to a glyphosate-based herbicide (GBH) or glyphosate alone (Gly) on female fertility and the hormonal and uterine milieu during the preimplantation period. F0 pregnant rats orally received a GBH or Gly in a dose of 2 mg of glyphosate/kg/day from gestational day (GD) 9 until weaning. F1 females were evaluated to determine the reproductive performance on GD19; and the sex steroid serum levels, the expression of estrogen receptor alpha (ERα), progesterone receptor (PR) and implantation-related genes on GD5 (preimplantation period). GBH and Gly induced preimplantation losses in F1 rats. GBH and Gly groups exhibited higher 17β-estradiol serum levels, without changes in progesterone. Both compounds increased the uterine ERα protein expression, with no differences at transcript level; and only Gly decreased PR mRNA expression. Also, GBH and Gly downregulated Hoxa10 and Lif genes, with no difference in Muc1 and Areg expression. To conclude, perinatal exposure to a GBH or Gly disrupted critical hormonal and uterine molecular targets during the receptive state, possibly associated with the implantation failures. Overall, similar results were found in GBH- and Gly-exposed rats, suggesting that the active principle might be the main responsible for the deleterious effects.

Brain-Selective Estrogen Therapy Prevents Androgen Deprivation-Associated Hot Flushes in a Rat Model.

Hot flushes are best-known for affecting menopausal women, but men who undergo life-saving castration due to androgen-sensitive prostate cancer also suffer from these vasomotor symptoms. Estrogen deficiency in these patients is a direct consequence of androgen deprivation, because estrogens (notably 17β-estradiol, E2) are produced from testosterone. Although estrogens alleviate hot flushes in these patients, they also cause adverse systemic side effects. Because only estrogens can provide mitigation of hot flushes on the basis of current clinical practices, there is an unmet need for an effective and safe pharmacotherapeutic intervention that would also greatly enhance patient adherence. To this end, we evaluated treatment of orchidectomized (ORDX) rats with 10β, 17β-dihydroxyestra-1,4-dien-3-one (DHED), a brain-selective bioprecursor prodrug of E2. A pilot pharmacokinetic study using oral administration of DHED to these animals revealed the formation of E2 in the brain without the appearance of the hormone in the circulation. Therefore, DHED treatment alleviated androgen deprivation-associated hot flushes without peripheral impact in the ORDX rat model. Concomitantly, we showed that DHED-derived E2 induced progesterone receptor gene expression in the hypothalamus without stimulating galanin expression in the anterior pituitary, further indicating the lack of systemic estrogen exposure upon oral treatment with DHED.

Molecular Evaluation of PROGINS Mutation in Progesterone Receptor Gene and Determination of its Frequency, Distribution Pattern and Association with Breast Cancer Susceptibility in Saudi Arabia.

Experimental and clinical evidence demonstrate that progesterone hormone and its nuclear receptor, the Progesterone Receptor (PR), play critical role in controlling mammary gland tumorigenesis and breast cancer development. Hormonal therapy (Tomaxifen) is the frontline treatment for hormone-dependent breast cancers. Progesterone hormone induces its action on the target cells by binding with its Progesterone receptor (PgR) therefore any genetic variations, which might induce alienation in the progesterone receptor, can result in an increased susceptibility of gynecological cancers. Alu insertion (PROGINS) mutation in PgR gene is reported to be associated with an increased risk of ovarian cancer and a decreased risk of breast cancer. However, its association with breast cancer risk remains inconclusive. Therefore, we investigated the association of PROGINS allele and its link with breast cancer risk. This case control study was performed on 200 subjects in which 100 were breast cancer cases and 100 gender matched healthy controls.The mutation was detected by using mutation specific PCR and results were confirmed by direct Sanger sequencing. A clinically significant difference was reported in genotype distribution of PROGINs allele among the cases and gender-matched healthy controls (P<0. 032). Genotype frequencies of A1/A1, A1/A2, A2/A2 reported in cases was 81%, 19% (18% & 1%) and in matched healthy controls were 93%, 7% (6% & 1%). The higher frequency of PROGINs allele (19%) was observed in cases than the healthy controls (7%). The findings indicated that PgR variants (CC vs CT) increased the risk of Breast cancer in codominant inheritance model with OR= 3.44, 95% CI =1. 30-9.09, P<0.021) whereas nonsignificant association was found for CC vs TT genotypes with OR=1.14, 95% CI=0.07-18.658, P=0. 92. However, subgroup analysis revealed that CT + TT vs CC genotype increased the risk of breast cancer in dominant inheritance model tested OR = 3. 11, 95% CI = (1.24-7.79), P = 0.015). A nonsignificant association for PgR (CC+CT) vs TT) genotypes were reported in breast cancer OR = 1. 0, 95% CI= (0. 061-16.21), P=1) in recessive inheritance model tested. However, analysis with clinicalpathological variables revealed that the PROGINs allele is significantly associated with the distant metastasis and advanced stage of the disease. The mutation specific PCR was successfully developed as an alternative to Sanger sequencing for the cost-effective detection for PROGINS allele of progesterone receptor gene. A clinically significant correlation of PROGINs allele was reported with the distant metastasis and advanced stage of the disease. Taken together, these results demonstrated that PROGINS variant is associated with an increased susceptibility to Breast cancer, providing novel insights into the genetic etiology and underlying biology of Breast carcinogenesis. Further studies with large sample sizes are required to validate our findings.

The Neandertal Progesterone Receptor.

The hormone progesterone is important for preparing the uterine lining for egg implantation and for maintaining the early stages of pregnancy. The gene encoding the progesterone receptor (PGR) carries introgressed Neandertal haplotypes with two missense substitutions and a mobile Alu element. These Neandertal gene variants have reached nearly 20% frequency in non-Africans and have been associated with preterm birth. Here, we show that one of the missense substitutions appears fixed in Neandertals, while the other substitution as well as the Alu insertion were polymorphic among Neandertals. We show that two Neandertal haplotypes carrying the PGR gene entered the modern human population and that present-day carriers of the Neandertal haplotypes express higher levels of the receptor. In a cohort of present-day Britons, these carriers have more siblings, fewer miscarriages, and less bleeding during early pregnancy suggesting that the Neandertal progesterone receptor alleles promote fertility. This may explain their high frequency in modern human populations.

Relaxed constraint and functional divergence of the progesterone receptor (PGR) in the human stem-lineage.

The steroid hormone progesterone, acting through the progesterone receptor (PR), a ligand-activated DNA-binding transcription factor, plays an essential role in regulating nearly every aspect of female reproductive biology. While many reproductive traits regulated by PR are conserved in mammals, Catarrhine primates evolved several derived traits including spontaneous decidualization, menstruation, and a divergent (and unknown) parturition signal, suggesting that PR may also have evolved divergent functions in Catarrhines. There is conflicting evidence, however, whether the progesterone receptor gene (PGR) was positively selected in the human lineage. Here we show that PGR evolved rapidly in the human stem-lineage (as well as other Catarrhine primates), which likely reflects an episode of relaxed selection intensity rather than positive selection. Coincident with the episode of relaxed selection intensity, ancestral sequence resurrection and functional tests indicate that the major human PR isoforms (PR-A and PR-B) evolved divergent functions in the human stem-lineage. These results suggest that the regulation of progesterone signaling by PR-A and PR-B may also have diverged in the human lineage and that non-human animal models of progesterone signaling may not faithfully recapitulate human biology.

Вовлеченность полиморфизма генов рецепторов эстрогенов и прогестерона в развитие миомы матки

Вовлеченность полиморфизма генов рецепторов эстрогенов и прогестерона в развитие миомы матки

SELECTION-GENETIC CHARACTERISTICS OF RABBITS POLTAVSKA SILVER BREED BY POLYMORPHISM OF PROGESTERONE RECEPTOR GENE

&#x0D; &#x0D; &#x0D; Genetic certification of the Poltavska silver rabbits by the progesterone receptor gene was carried out by amplification of the corresponding sections of genomic DNA in the Polymerase Chain Reaction (PCR). As a result of PCR analysis it was found that the discrete DNA bands of rabbits were intensive hybridization spectra and characterized by different numbers, locations, and intensities of detected fragments. Based on the results of genotyping of Poltavska silver rabbits the following data on their genetic structure were obtained for gene polymorphism G2464A. In actual distribution of genotypes number of homozygous animals with the G gene was 18.3% (11 animals), number of homozygotes for A allele was almost twice as large (31.7%). The Wright fixation index for both polymorphic variants was negative which demonstrates the advantage of heterozygotes in population for tested genes. It was established that high values of multiple fertility rate in three aprons were found in rabbits with the GG genotype, and the lowest - with the AA genotype.&#x0D; In general there was a tendency to increase fertility by an average of 12% in three spouts for the “desired genotype” GG compared to animals that had genotypes AA and AG (p&lt;0.05). Analysis of variation in milk yield of rabbits genotypes by polymorphic variants of progesterone receptor showed that this indicator was higher in homozygous animals AA, and the lowest - in heterozygotes AG. Moreover, rabbits with GG genotype were 3% inferior to them (the difference is not significant). According to the results of one-way analysis of variance, the influence of the rabbit genotype on the progesterone receptor gene on their milk production was found but no statistically significant difference between groups of animals with different genotypes. According to the data obtained it can be argued that the genotype of Poltavska Silver female rabbits in progesterone receptor gene has statistically significant effect on the manifestation of multiple pregnancy which can also be used in breeding.&#x0D; &#x0D; &#x0D;

A modern look at the molecular-biological mechanisms of breast tumours in dogs

High morbidity and increase in the number of registrations of breast tumours in dogs, their wide application as biological models, discussion of numerous questions of oncogenesis, and the lack of a uniform/unified methodological approach to the study of molecular and biological mechanisms of treatment of cancer determine the relevance of the problem of cancer both in humans and in our domestic companions. The analysis of publications allowed us to establish the following patterns of carcinogenesis. The peculiarities of the biological behaviour of breast tumours depend on their pathomorphological structure. Genetic predisposition to breast cancer is characteristic only in the single breed aspect. Environmental factors are of critical relevance to carcinogenesis : chemical pollutants initiate oncogenesis indirectly – by altering the expression of several receptors, impaired endocrine balance and direct mutagenic effects. Reproductive status plays a key role in the initiation and progression of breast tumours by reducing the expression of estrogen, progesterone and prolactin receptor genes. The inflammatory response that accompanies the neoplasia process is characterized by increased production of cytokines, cyclooxygenase-2, interleukins (IL-1, IL-6, IL-8), vascular endothelial growth receptors, and impaired hemostatic status (oxidative stress), which promotes progression of disease. In breast cancer in dogs, genomic instability leads to genomic aberrations, and subsequently, mutations that support the proliferation, survival and dissemination of neoplastic cells. The initiation and progression of mammary gland tumours is provided by cancer stem cells by disrupting the regulation of precursor cell self-renewal, which also predispose to resistance to chemotherapeutic agents, radiation, and hormonal cancer therapy. The analysis of the publications revealed the major markers of carcinogenesis that could potentially be used as biological targets for the design of modern diagnostic strategies and high-performance therapeutic protocols.

Structural characterization, antioxidant and cytotoxic effects of iron nanoparticles synthesized using Asphodelus aestivus Brot. aqueous extract

AbstractASPwas used to synthesize FeNPA. They were characterized by UV-vis spectroscopy, FT-IR, TEM, SEM, XRD and ZP. The aim of this study was to evaluate in vitro cytotoxic activity and antioxidant acitivities of FeNPAandASP. The antioxidant properties were evaluated using DPPH, ABTS+and H2O2assays. FeNPAhad higher antioxidant activity comparing toASPaccording to DPPH (IC50: 3.48 μg/mL) and ABTS+(60.52%) assays. Anti-cancer activities of FeNPAandASPwere investigated in breast cancer, melanoma and control cell lines. FeNPAwas more cytotoxic thanASPin MCF-7, MeWo, CHL-1, and HEL 299 cells. FeNPAhad shown that mitochondria induce apoptosis through stress in MDA-MB-231, and cells MeWo.ASPalso induced apoptosis 2.23-fold in MCF-7 cells. Progesterone receptor gene expression showed a 10-fold increase in a hormone-dependent MCF-7 cell line inASP, and FeNPAtreatment. Expressions of BCL6, CXCL12, DNAJC15, RB1 and TPM1 in melanoma cancer cell lines were significantly increased inASPand FeNPAadministration. It had been shown that FeNPAregulates gene expressions that may be considered important in terms of prognosis in breast cancer and melanoma cell lines and it is suggested that gene expressions regulated by FeNPAare also evaluated in animal models in vivo.

Progesterone Receptor Gene Variants in Metastatic Estrogen Receptor Positive Breast Cancer.

Tumor mutations in the gene encoding estrogen receptor alpha (ESR1) have been identified in metastatic breast cancer patients with endocrine therapy resistance. However, relatively little is known about the occurrence of mutations in the progesterone receptor (PGR) gene in this population. The study objective was to determine the frequency and prognostic significance of tumor PGR mutations for patients with estrogen receptor (ER)-positive metastatic breast cancer. Thirty-five women with metastatic or locally recurrent ER+ breast cancer were included in this IRB-approved, retrospective study. Targeted next-generation sequencing of the PGR gene was performed on isolated tumor DNA. Associations between mutation status and clinicopathologic factors were analyzed as well as overall survival (OS) from time of metastatic diagnosis. The effect of the PGR variant Y890C (c.2669A>G) identified in this cohort on PR transactivation function was tested using ER-PR- (MDA-MB-231), ER+PR+ (T47D), and ER+PR- (T47D PR KO) breast cancer cell lines. There were 71 occurrences of protein-coding PGR variants in 67% (24/36; 95% CI 49-81%) of lesions. Of the 49 unique variants, 14 are single nucleotide polymorphisms (SNPs). Excluding SNPs, the median OS of patients with PGR variants was 32months compared to 79months with wild-type PGR (p = 0.42). The most frequently occurring (4/36 lesions) non-SNP variant was Y890C. Cells expressing Y890C had reduced progestin-stimulated PR transactivation compared to cells expressing wild-type PR. PGR variants occur frequently in ER+ metastatic breast cancer. Although some variants are SNPs, others are predicted to be functionally deleterious as demonstrated with Y890C PR.

Mutations in the Progesterone Receptor (PROGINS) May Reduce the Symptoms of Acute Hepatitis E and Protect Against Infection.

BackgroundMutations in the progesterone receptor (PR) gene, PROGINS, have been studied in relation to hepatitis E virus (HEV) infection. Patients with the PROGINS gene may develop a worse clinical course of hepatitis E. The aim of our study was to evaluate the influence of PROGINS on the susceptibility to and the clinical course of HEV infection in HIV patients.MethodsThis study included patients with HIV who were evaluated in previous prospective studies for the prevalence and incidence of HEV. The following three groups of patients were studied: (i) never infected, (ii) past infections, and (iii) recently infected. We determined the PR genotype to evaluate the proportion of patients who were homozygous for PROGINS according to HEV infection. We also compared the proportion of PROGINS carriers with a recent HEV infection according to their symptomatology.ResultsIn this study, 311 patients infected with HIV were included. Of those patients, 198 were homozygous wild type (63.7%), 91 were heterozygous (29.3%), and 22 were homozygous PROGINS (7.1%). We found that the homozygous PROGINS genotype in women was associated with a lower HEV seroprevalence. In addition, in patients with a recent HEV infection, none of those homozygous for PROGINS presented symptoms.ConclusionThe PROGINS mutation plays a protective role against HEV infection and is associated with subclinical infection in HIV-infected patients, particularly women.

Dimethyl sulfoxide maintains structure and function of cryopreserved equine endometrial explants

Availability of viable frozen-thawed endometrial tissues could facilitate detailed studies into physiologic and disease processes influencing the endometrium. This study was designed to investigate the cryosurvival of equine endometrial tissue. Previous studies in the human and horse have focused on cryopreservation of dissociated endometrial cells. To our knowledge, there are no studies on cryopreservation of endometrial explants. Our objectives were to 1) determine the influence of differing concentrations of the permeating cryoprotectant dimethyl sulfoxide (Me2SO) on viability, structural integrity, and gene expression of cryopreserved equine endometrial tissues prior to and following a 5-day explant culture in vitro and 2) examine the influence of low (1000 mg/L dextrose) vs high (4500 mg/L dextrose) glucose medium during in vitro culture. Both 10% and 20% (v/v) concentrations of Me2SO maintained viability following cryopreservation and in vitro culture. In addition, gene expression remained unaltered following cryopreservation with either 10% or 20% Me2SO. However, tissue structural integrity was slightly reduced compared to the fresh control. Furthermore, there was no difference in structural integrity, cell viability, or gene expression between low and high glucose medium during in vitro culture. Although E-cadherin and Ki67 gene expression was not different among fresh, 10% Me2SO, and 20% Me2SO treatments prior to or following tissue culture, estrogen receptor-α and progesterone receptor gene expression were reduced in all groups after explant culture. This is the first report of successful cryopreservation of equine endometrial explants.

Effects of COH on the expression of connexin43 in endometrial stromal cells

ObjectiveThe purpose of this study was to investigate the effect of controlled ovarian hyperstimulation (COH) on gap junction, and to induce the effect of an estrogen level overdose on gap junction in vitro by COH. Here, we mainly focus on connexion43 (Cx43), progesterone receptor (PR) and prolactin-related protein (PRP), and CyclinD3 genes expression, as well as the expression of Cx43 protein, were investigated. Materials and methodsMature BDF-1 mice were divided into different COH, and the mouse uterus was isolated, Paraffin sections evaluate the effect of COH on mouse uterine endometrial morphology. The other part was used for the extraction of mouse uterine endometrial stromal cells (ESC), some related gene changes are detected. Human ESC were isolated from human endometrium by primary culture, the estrogen concentrations 10−6 mol/L, 10−7 mol/L were added, the changes of Cx43 gene and related proteins were detected, too. Results(1) HE staining showed that in the ovulatory endometrium of mice in the high super ovulation group, uterine glands in the stromal layer were significantly increased, the relative vascular tissues was less abundant. (2) In three groups of COH mice, the expression of Cx43, PR, and PRP genes in ESC was significantly different (P < 0.05). (3) In vitro ESC in the COH group showed significant differences in Cx43, PR, and CyclinD3 gene expression (P < 0.05), and showed an obvious dose effect. In addition, Western blot analysis showed that the Cx43 protein and Cx43 gene expression were similar. Conclusions(1) Animal experiments study showed that Cx43 gene expression in ESC was significantly decreased in hyper COH, in addition, the advance in gene expression was significantly earlier, suggesting decidualization appeared significantly earlier. (2) In vitro COH demonstrated when the estrogen concentration used was higher, the expression level of Cx43 gene and protein was lower. Combined with animal experiments, the endometrium decidualization was advanced in mice that were underwent hyper COH, which may reflect the endometrial receptivity.

Primary endometrial 3D co-cultures: A comparison between human and rat endometrium

Human and rat reproductive systems differ significantly with respect to hormonal cyclicity and endometrial cell behavior. However, species-differences in endometrial cell responses upon hormonal stimulation and exposure to potentially toxic compounds are poorly characterized. In this study, human and rat endometrial hormonal responses were assessed in vitro using a 3D co-culture model of primary human and rat endometrial cells. The models were exposed to the aryl hydrocarbon receptor (AHR) ligands 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), laquinimod, and its AHR active metabolite DELAQ.In both the human and rat endometrial models, estrogen receptor and progesterone receptor gene expression was modulated by the hormonal treatments, comparable to the in vivo situation. AHR gene expression in the human endometrial model did not change when exposed to hormones. In contrast, AHR expression decreased 2-fold in the rat model when exposed to predominantly progesterone, which resulted in a 2.8-fold attenuation of gene expression induction of cytochrome P450 1A1 (CYP1A1) by TCDD. TCDD and DELAQ, but not laquinimod, concentration-dependently induced CYP1A1 gene expression in both human and rat endometrial models. Interestingly, the relative degree of DELAQ to induce CYP1A1 was higher than that of TCDD in the human model, while it was lower in the rat model.These data clearly show species-differences in response to hormones and AHR ligands between human and rat endometrial cells in vitro, which might greatly affect the applicability of the rat as translational model for human endometrial effects. This warrants further development of human relevant, endometrium-specific test methods for risk assessment purposes.

Tissue-specific progesterone receptor-chromatin binding and the regulation of progesterone-dependent gene expression

Progesterone receptor (PGR) co-ordinately regulates ovulation, fertilisation and embryo implantation through tissue-specific actions, but the mechanisms for divergent PGR action are poorly understood. Here we characterised PGR activity in mouse granulosa cells using combined ChIP-seq for PGR and H3K27ac and gene expression microarray. Comparison of granulosa, uterus and oviduct PGR-dependent genes showed almost complete tissue specificity in PGR target gene profiles. In granulosa cells 82% of identified PGR-regulated genes bound PGR within 3 kb of the gene and PGR binding sites were highly enriched in proximal promoter regions in close proximity to H3K27ac-modified active chromatin. Motif analysis showed highly enriched PGR binding to the PGR response element (GnACAnnnTGTnC), but PGR also interacted significantly with other transcription factor binding motifs. In uterus PGR showed far more tendency to bind intergenic chromatin regions and low evidence of interaction with other transcription factors. This is the first genome-wide description of PGR action in granulosa cells and systematic comparison of diverse PGR action in different reproductive tissues. It clarifies finely-tuned contextual PGR-chromatin interactions with implications for more targeted reproductive medicine.

Endometrial luminal epithelial cells sense embryo elongation in the roe deer independent of interferon-tau†.

Numerous intrauterine changes take place across species during embryo development. Following fertilization in July/August, the European roe deer (Capreolus capreolus) embryo undergoes diapause until embryonic elongation in December/January. Embryonic elongation prior to implantation is a common feature among ungulates. Unlike many other ruminants, the roe deer embryo does not secrete interferon-tau (IFNτ). This provides the unique opportunity to unravel IFNτ-independent signaling pathways associated with maternal recognition of pregnancy (MRP). This study aimed at identifying the cell-type-specific endometrial gene expression changes associated with the MRP at the time of embryo elongation that are independent of IFNτ in roe deer. The messenger RNA (mRNA) expression of genes known to be involved in embryo-maternal communication in cattle, pig, sheep, and mice was analyzed in laser capture microdissected (LMD) endometrial luminal, glandular epithelial, as well as stromal cells. The mRNA transcript abundances of the estrogen (ESR1), progesterone receptor (PGR), and IFNτ-stimulated genes were lower in the luminal epithelium in the presence of an elongated embryo compared to diapause. Retinol Binding Protein-4 (RBP4), a key factor involved in placentation, was more abundant in the luminal epithelium in the presence of an elongated embryo. The progesterone receptor localization was visualized by immunohistochemistry, showing an absence in the luminal epithelium and an overall lower abundance with time and thus prolonged progesterone exposure. Our data show a developmental stage-specific mRNA expression pattern in the luminal epithelium, indicating that these cells sense the presence of an elongated embryo in an IFNτ-independent manner.

Genomic sequence analyses of classical and non-classical lamprey progesterone receptor genes and the inference of homologous gene evolution in metazoans

BackgroundNuclear progesterone receptor (nPR) is an evolutionary innovation in vertebrates that mediates genomic responses to progesterone. Vertebrates also respond to progesterone via membrane progesterone receptors (mPRs) or membrane associated progesterone receptors (MAPRs) through rapid nongenomic mechanisms. Lampreys are extant agnathan vertebrates, residing at the evolutionary juncture where vertebrates diverged from invertebrates. A survey of the progesterone receptor (PR) gene sequences in lamprey genomes would inform PR gene evolutionary events during the transition from invertebrates to vertebrates.ResultsIn this study, we annotated sequences of one nPR, four mPR (β, γ, δ and ε) and four MAPR genes from genomes of two lamprey species (Petromyzon marinus and Lethenteron japonicum). To infer the origin and evolutionary history of PR genes, we constructed phylogenetic trees of PR homologous sequences across representative species of metazoans. Phylogenetic analyses revealed that the mPRγ gene first appeared in non-bilaterians, and the mPRβ gene likely arose from a duplication of mPRγ. On the other hand, the mPRγ gene gave rise to the mPRδ and ε genes much later in the vertebrate lineage. In addition, the mPRα gene first appeared in cartilaginous fishes, likely derived from duplication of mPRβ after the agnathan-gnathostome divergence. All known MAPR genes were present in the lamprey genomes. Progesterone receptor membrane component 1 (PGRMC1), neudesin and neuferricin genes probably evolved in parallel in non-bilaterians, whereas two copies of PGRMC genes probably derived from duplication of ancestral PGRMC1 sequence and appeared before the speciation of lampreys.ConclusionsNon-classical mPR and MAPR genes first evolved in non-bilaterians and classical nPR genes evolved later in basal vertebrates. Sequence repertoires for membrane progesterone receptor genes in vertebrates likely originated from an ancestral metazoan sequence and expanded via several duplication events.