Kruppel-like factor 5 (Klf5) in fetal-maternal tissue during periimplantation and effects of ovarian steroid hormone antagonist on its expression during uterine receptivity of albino mice

Background Embryo implantation is a tightly regulated sequence of events regulated by ovarian steroids, estrogen and progesterone, and their downstream targets. Ovarian steroids regulate most of the genes involved in embryo implantation and pregnancy. However, some factors are not regulated by ovarian steroids, estrogen, progesterone, or both. Kruppel-like factor 5 (Klf5) is an example of an ovarian steroid–independent factor having a role in cellular proliferation, differentiation. The detailed expression profile of Klf5 during uterine receptivity and periimplantation has not been studied till now. In the present research work, an attempt was made to investigate the expression pattern of Klf5 in mice fetal-maternal tissue during periimplantation (day 4–day 8). The expressional and functional independence of Klf5 on the ovarian steroids was studied using estrogen and progesterone antagonist. The study was carried out in female Swiss albino mice of LACA strain during the periimplantation period. KLF5 was localized in the fetal-maternal tissues using the immunofluorescence technique in paraffin-embedded tissues. Ovarian steroid antagonists were administered subcutaneously from day 1 to day 3 of gestation, and the uterus was collected on the morning of day 4. Klf5 protein and mRNA levels were studied by western blot and quantitative real-time PCR (qPCR), respectively. Results KLF5 was localized in the embryo, uterine luminal epithelium, glandular epithelium, and proliferating stromal cells during periimplantation. In ovarian steroid antagonist–treated groups, KLF5 was localized in the luminal and glandular epithelium and stroma. Western blot and qPCR confirmed translation and transcription of KLF5 during the experimental period. The KLF5 protein level significantly increased on day 6, day 7, and day 8 when compared with day 4 (P < 0.05). The mRNA level of Klf5 increased significantly on day 7 and day 8 when compared with day 4 (P < 0.05). In ovarian steroid antagonist–treated groups, protein and mRNA corresponding to Klf5 were observed. From this finding, it can be assumed that Klf5 may be a steroid-independent factor expressed during uterine receptivity. Conclusion Spatiotemporal KLF5 expression in fetal-maternal tissue was observed during the experimental period. The results suggest that Klf5 is an ovarian steroid–independent factor that may play a pivotal role in implantation, decidualization, and embryogenesis.

Unique transcriptomic changes underlie hormonal interactions during mammary histomorphogenesis in female pigs

Successful lactation and the risk for developing breast cancer depend on growth and differentiation of the mammary gland (MG) epithelium that is regulated by ovarian steroids (17beta-estradiol [E] and progesterone [P]) and pituitary-derived prolactin (PRL). Given that the MG of pigs share histomorphogenic features present in the normal human breast, we sought to define the transcriptional responses within the MG of pigs following exposure to all combinations of these hormones. Hormone-ablated female pigs were administered combinations of E, medroxyprogesterone 17-acetate (source of P), and either haloperidol (to induce PRL) or 2-bromo-α-ergocryptine. We subsequently monitored phenotypic changes in the MG including mitosis, receptors for E and P (ESR1 and PGR), level of phosphorylated STAT5 (pSTAT5), and the frequency of terminal ductal lobular unit (TDLU) subtypes; these changes were then associated with all transcriptomic changes. Estrogen altered the expression of ~20% of all genes that mostly associated with mitosis, whereas PRL stimulated elements of fatty acid metabolism and an inflammatory response. Several outcomes, including increased pSTAT5, highlighted the ability of E to enhance PRL action. Regression of transcriptomic changes against several MG phenotypes revealed 1,669 genes correlated with proliferation, among which 29 were E-inducible. Additional gene expression signatures were associated with TDLU formation and the frequency of ESR1 or PGR. These data provide a link between the hormone-regulated genome and phenome of the MG in a species having a complex histoarchitecture like that in the human breast, and highlight an underexplored synergy between the actions of E and PRL during MG development.

Subgenual cingulate resting regional cerebral blood flow in premenstrual dysphoric disorder: differential regulation by ovarian steroids and preliminary evidence for an association with expression of ESC/E(Z) complex genes

Substantial evidence suggests that circulating ovarian steroids modulate behavior differently in women with PMDD than in those without this condition. However, hormonal state-related abnormalities of neural functioning in PMDD remain to be better characterized. In addition, while altered neural function in PMDD likely co-exists with alterations in intrinsic cellular function, such a relationship has not been explored. Here, we investigated the effects of ovarian steroids on basal, resting regional cerebral blood flow (rCBF) in PMDD, and, in an exploratory analysis, we tested whether the rCBF findings were linked to the expression of ESC/E(Z) genes, which form an essential ovarian steroid-regulated gene-silencing complex. Resting rCBF was measured with oxygen-15 water PET (189 PET sessions in 43 healthy women and 20 women with PMDD) during three self-as-own-control conditions: GnRH agonist (Lupron)-induced ovarian suppression, estradiol add-back, and progesterone add-back. ESC/E(Z) gene expression data were obtained from RNA-sequencing of lymphoblastoid cell lines performed in a previous study and were examined in relation to hormone-induced changes in rCBF. In the rCBF PET data, there was a significant diagnosis-by-hormone interaction in the subgenual cingulate (PFDR = 0.05), an important neuroanatomical hub for regulating affective state. Whereas control women showed no hormonally-related changes in resting rCBF, those with PMDD showed decreased resting rCBF during both estradiol (P = 0.02) and progesterone (P = 0.0002) add-back conditions. In addition, in PMDD, ESC/E(Z) gene expression correlated with the change in resting rCBF between Lupron-alone and progesterone conditions (Pearson r = −0.807, P = 0.016). This work offers a formulation of PMDD that integrates behavioral, neural circuit, and cellular mechanisms, and may provide new targets for future therapeutic interventions.

Protective Effect of 17β Estradiol against Kidney Injury in a Mouse Model of Unilateral Renal Ischemia

Renal ischemia can lead to irreversible damage to the kidney including atrophy and fibrosis. Young females generally experience attenuated pathology due to the protective actions of ovarian steroids, in particular 17β-Estradiol (E2); however, the mechanisms of action remain murky.

Steroid regulation of secreted phosphoprotein 1 (SPP1) expression in ovine endometrium

Secreted phosphoprotein 1 (SPP1) is an extracellular matrix glycoprotein that is highly expressed at the maternal–fetal interface and is a critical mediator of embryo implantation. The objectives of this study were to examine the spatial and temporal cyclical expression patterns and steroid regulation of SPP1 mRNA and protein in ovine endometrium, which may be further indicative of their functionality in embryo implantation. Uterine tissue was obtained following hysterectomy from ovariectomised ewes treated with ovarian steroids. In parallel, invitro culture of endometrial cells was used to investigate the effects of ovarian steroids on SPP1 expression in endometrial and luminal epithelial (LE) cells. A significant sustained mid-luteal phase increase in SPP1 mRNA in intercaruncular regions of the endometrium was observed, indicating that glandular epithelium is likely to be the primary source of SPP1 production. This increase in SPP1 was induced by progesterone treatment and was shown at the protein level by immunohistochemistry analysis. Similarly, treatment of stromal cells with 10ng mL−1 progesterone or in combination with 1ng mL−1 oestradiol significantly increased SPP1 expression (P&lt;0.05). Collectively, expression levels of SPP1 are cycle-dependent and peak in the progesterone-dominant luteal phase. They are dependent on the interaction of uterine LE and stromal cells and may involve paracrine signalling by progesterone receptor-positive stromal cells.

THE INFLUENCE OF HYPERPROLACTINEMIA ON THE LEVEL OF ESTRADIOL AND PROGESTERONE IN WOMEN

Prolactin is a polypeptide hormone that is synthesized and secreted by specialized cells in the adenohypophysis called lactotrophs.Excretion of prolactin from lactotrophs is regulated by neurohormones or neurotransmitters who are secreted in the hypothalamus, and they have a stimulating or inhibiting role in its secretion.By maintaining a balance in the excretion of these factors, the level of prolactin concentration in the serum is within normal reference limits.Disruption of dopamine excretion as the strongest inhibitor of prolactin secretion increases the level of prolactin, a condition known as hyperprolactinaemia. A prolonged increase in prolactin levels in serum, in the absence of pregnancy or lactation is referred to as pathological hyperptopectinemia, which is most common in women. Physiological conditions in which an increased level of prolactin appears, and which are not a disease, are pregnancy, breast-feeding, nipple stimulation, physical activity, stress condition, longer-term sleep, and more.The aim of the study was to determine the level of prolactin in the group of patients with confirmed hyperprolactinaemia, compared with the control group of patients, and at the same time to analyze the influence of hyperprolactinemia on the level of ovarian steroids, estradiol, and progesterone in the hyperprolactinemic group of patients,in relation to the control group. Patients aged 25-35 years with pre-established hyperprolactinaemia were examined. Prolactin, estradiol and progesterone were examined. Patients were divided into three groups, a control group of patients with values for prolactin within the limits of the reference, a group of patients with diagnosed hyperprolactinaemia,that is an elevated level of prolactin and a group under the therapy with Bromergon or Dostinex.Our results suggest that in hyperprolactinemia the concentration of prolactin in patients is significantly increased in relation to the control group, but after treatment with Dostinex or Bromergon in the treated group of patients, the level of prolactin significantly decreases to concentrations close to the control group. In hyperprolactinemic patients, the serum estradiol level was significantly reduced with respect to the control group, but with the application of antiprolactin therapy the concentration significantly increased to levels close or higher compared to the control group. The value of progesterone in hyperprolactinemic patients is significantly reduced in relation to the control group, but in the treated group of patients with Bromergon or Dostinex there is a significant increase in progesterone in the serum to levels that are slightly higher compared to the values for progesterone from the control group.