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Realization of estradiol effects in the uterus of ovariectomized rats under acute stress.

The aim of this study was to examine the influence of acute stress on the realization of the effects of estrogens in the uterus of ovariectomized rats. The rats were immobilized for 6 h and given a single injection of estradiol dipropionate (E2; 10 micrograms/rat, i.m.). E2-only treated rats, olive oil injected and stressed rats, olive oil-only treated rats and untreated ovariectomized animals were used as controls. The effect of E2 was assessed by the volumes of cells, nuclei and nucleouli (morphometry), DNA content (Feulgen technique) and proliferative activity (mitotic index) in luminal epithelium, in glandular epithelium and in stromal cells of endometrium 24, 36 and 48 h after the injection of E2 or olive oil. All the effects of E2 were reduced in E2-treated rats subjected to stress in all the uterine structures. In olive oil treated rats, stress induced certain increase in the volume of cells and nuclei and slight increase in the DNA content in all the structures. It is suggested that the influence of stress on the uterus is mediated by impairment of some septs in the mechanism of action of estrogen on the uterus, which leads to a decrease in the sensitivity of the uterine structures to estrogens. Without estrogen stimulation the effect of stress on the uterus is realized via estrogen-independent pathways.

Animals↗

Effect of estradiol on prostaglandin metabolism in rat uterus.

The purpose of this work was to investigate the effect of a single injection of estradiol (10 microng/rat) on prostaglandin(PG) metabolism in rat uterus. The PG content of uterus was increased within 6 hr after estradiol injection but the incorporation of 3H-arachidonic acid into PG in uterus was not changed within same period. The uptake of 3H-PG was increased within 6 hr after estradiol injection. In these results, it is discussed that PG from other tissue are also involved in the metabolism of PG in uterus.

Animals↗

Localization of bradykinin B2 receptors in sheep uterus.

The localization and characterization of bradykinin B2 receptors in sheep uterus was carried out using a radiolabelled B2 receptor ligand, 3,4-hydroxyphenypropionyl-D-Arg0-[Hyp3,Thi5,D-++ +Tic7,D-Oic8] bradykinin (HPP-HOE140). Competition of the radioligand, [125I]HPP-HOE140, from membrane preparations of anoestrus sheep uterus by bradykinin agonists and antagonists revealed the presence of high- and low-affinity binding sites with ligand specificity typical of the bradykinin B2 receptor. Using in vitro autoradiography on tissue sections, intense binding was visible over the superficial epithelial layer of the endometrium and inner third of the myometrium of anoestrus sheep uterus. Bradykinin B2 receptors in the myometrium were down regulated in pregnant sheep uterus. We demonstrate that binding studies using [125I]HPP-HOE140 offer high sensitivity and specificity for characterization, quantitation and localization of the bradykinin B2 receptors in tissues and offers new information on uterine bradykinin B2 receptors.

Anestrus↗

Isolation and localization of neurophysin-like proteins in rat uterus.

Neurophysins are part of the prohormones for vasopressin and oxytocin, and are localized with these hormones in the magnocellular cells of the neurohypophysis. New techniques have identified neurophysins in other areas within and outside the central nervous system, and we report here the isolation of neurophysins from the uterus of the rat. Using immunohistology the neurophysin immunoreactivity was localized to the epithelial lining cells of the uterus, and using radioimmunoassay was also present in uterine fluid suggesting secretion into the uterine cavity. The amount of uterine neurophysin increased in response to administered estrogen and was especially elevated in the pregnant uterus. The neurophysin-like material isolated from the uterus was similar to neurophysins from the neurohypophysis by radioimmunoassay, molecular sieve chromatography, isoelectric focusing and SDS gel electrophoresis. Both neurohypophyseal hormones, vasopressin and oxytocin, were also extracted from uterine endothelium and identified by radioimmunoassay and high pressure liquid chromatography.

Animals↗

Effect of oestradiol and progesterone on prostaglandin production by the rabbit uterus in vitro.

Endogenous concentrations of prostaglandin (PG) F2 alpha and PGE2, but not of 6-oxo-PGF2 alpha, in the uterus were significantly higher in ovariectomised, control rabbits than in ovariectomised rabbits which received oestradiol and/or progesterone treatment. However, due to significant increases in uterine weight following steroid treatment, there were no significant differences in the total uterine content of PGF2 alpha or PGE2 following hormonal treatment, whereas total 6-oxo-PGF1 alpha content in the uterus was significantly increased after oestradiol treatment when given alone or following progesterone priming. When PG synthesis by uterine homogenates in vitro was examined, expressing the results on a unit weight basis showed that oestradiol treatment alone significantly depressed the production of PGF2 alpha and PGE2, but not of 6-oxo-PGF1 alpha, when compared to ovariectomised, control rabbits. Progesterone treatment had no significant effect, and reversed the effect of oestradiol on PGF2 alpha synthesis. When the results were expressed on a total uterine basis, however, oestradiol, acting on a progesterone primed uterus, was the optimum stimulus for maximum PGF2 alpha synthesis. Total PGE2 production was not significantly affected by steroidal treatment, whereas oestradiol, alone or following progesterone priming, significantly increased 6-oxo-PGF1 alpha production. These findings are discussed in relation to the stimulus necessary for increased PGF2 alpha secretion from the rabbit uterus at the end of pseudopregnancy.

Animals↗

The role of prolactin in realization of estradiol action in the uterus of ovariectomized rats.

The aim of this study was to examine the role of prolactin (PRL) in promoting the effects of estradiol (E2) on the uterus. Ovariectomized rats were given a single injection of E2 dipropionate (10 micrograms/rat), along with injections of ovine prolactin (PRL; a single dose--2.5 IU/rat), or injections of metoclopramide (a single dose--0.5 mg/rat), or injections of bromocriptine (a single dose--0.25 mg/rat). The effects of E2 on the uterus were determined by measuring mitotic indexes; cell, nucleus and nucleolus volumes (morphometry), and DNA content (Feulgen's method) in luminal and glandular epithelia, endometrial stromal cells at 24, 36 and 48 h after the injection of E2. It is revealed that at each time point after E2 and PRL or metoclopramide treatment almost all the tested parameters were lower than those in control rats treated with E2 and distilled water. The values obtained in all the periods after E2 along with bromocriptine treatment were higher compared with those in E2-treated animals. Most of the results are statistically significant (P < 0.05-0.001). In the absence of E2 none of the drugs used influenced the uterine parameters tested. The effect of PRL is likely to be mediated by their receptors in the uterus, and it occurs by changing the mechanism of estrogen action in the uterus. The actions of metoclopramide and bromocriptine are probably also mediated by PRL.

Analysis of Variance↗

Calbindin-D9k gene expression during the perinatal period in the rat: correlation to estrogen receptor expression in uterus.

Calbindin-D9k (CaBP-9k) is a cytosolic calcium binding protein mainly expressed in duodenum, placenta and uterus. The gene encoding the rat CaBP-9k is subject to tissue specific induction by 1,25 dihydroxyvitamin D3 (intestine) and estradiol (E2) (uterus). Control of placental expression remains unknown. The expression of CaBP-9k mRNA during the perinatal period was studied (pregnancy day 21 (P21)-lactation day 4 (L4)). In uterus, maximal expression levels were found at P21 and maintained until L1. With the transition to L2, the CaBP-9k mRNA concentration dropped drastically below the detection limit as quantitated by Northern blot analysis. Measurements of E2 and progesterone (P) levels showed a gradual decrease at late pregnancy (P21; birth). Post partum E2 levels continued to decline and P concentrations increased slightly. Uterine estrogen receptor (ER) mRNA levels determined by cDNA/PCR analysis revealed close correlation between expression of ER and CaBP-9k mRNAs. ER mRNA levels were maximal at P22 and declined at parturition and with onset of lactation. At L2 and L3 ER mRNA levels were minimal and had decreased 5-fold compared to late pregnancy. CaBP-9k protein concentrations fluctuated only slightly dependent on the stage of the estrous cycle: estrus > proestrus > diestrus. During the perinatal period CaBP-9k concentration was overall lower than in non-pregnant uterus and revealed only a moderate increase at birth and decrease in early lactation. Similar to the uterine levels, placental CaBP-9k mRNA was highest at P21 and remained high until birth. Fetal duodenal CaBP-9k rose sharply just prior to birth and plateaued in the early postpartal period.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Expression of pituitary peptide 23 in the rat uterus: regulation by estradiol.

Peptide 23 was first identified in pituitary cell conditioned medium as a secreted protein which was regulated in a similar fashion to growth hormone. It was subsequently found to be a member of the C-type lectin gene superfamily and identical to pancreatis associated protein (PAP). It is widely expressed in the gastrointestinal tract. Our present study demonstrates that peptide 23 gene is also expressed in the uterus. Peptide 23/PAP mRNA was at highest levels during estrus and was not detectable in the immature rat uterus. A single injection of 17 beta-estradiol resulted in a transient induction of peptide 23/PAP mRNA in ovariectomized rats whereas a sustained induction was seen with diethylstilbestrol implants. In situ hybridization localized peptide 23/PAP mRNA to the luminal epithelial cells. During gestation, peptide 23/PAP mRNA was detected only in the uterine samples from day 12 to 18 of pregnancy with maximal expression on day 12. Peptide 23 expression was confined to the uterus itself and not expressed in either the decidua or the fetal tissues. PSP/reg, another closely related member of the C-lectin gene family was not expressed in any of these uterine tissues. These results indicate that estrogen may act as a physiological regulator of peptide 23 in the uterus and suggests that this protein may have some role in estrogen action.

Animals↗

Swine uterus carnosinase activity in oestrous cycle and early pregnancy.

Carnosinase activity was determined in uterus extracts of sexually immature sows, on particular days of the oestrous cycle, and on the 20th and 30th day of pregnancy. In mature sows carnosinase activity in the uterus was on the average 4.5 times higher than in immature sows. Activity of the enzyme in the oestrous cycle increased from the zero day (first day of the heat) until 13th day, followed by a rapid decrease, reaching the lowest levels on the 17th day of the cycle (3 times lower on the average than on the zero day). On the last days of the cycle (20-21st) activity of carnosinase reached again levels similar to those of the zero day. Carnosinase activity in a uterus corner of pregnant sows (20th day of pregnancy) was over 4 times higher than in the "peak" day of the oestrous cycle (13th day), and over 12 times higher than in immature sows. Activity of the enzyme increased along with progressing pregnancy. It was found that activity of carnosinase in uterus corner of swines was related to the level of progesterone determined by other authors in the blood plasma.

Animals↗

The effects of hormone substitution in depot form on the uterus in a group of 50 perimenopausal women--a vaginosonographic study.

The aim of our study was to examine the effects of hormone replacement on the size of the uterus and the development or increase of myomatas. Fifty perimenopausal women were included in the study (53.8 +/- 5.0 years). Patients received a substitution therapy composed of a combination of 4 mg estradiovalerate and 200 mg prasteronenantate (Gynodian Depot cartridges) given as a muscular injection in 6-10 week intervals (mean 7 weeks +/- 4 days). Prior to the onset of therapy with Gynodian and after a period of 12 months (+/- 13 days) vaginosonography was performed. Measurements taken were length, thickness, height of endometrium, size of ovaries and of myomas. Data obtained were correlated with baseline findings. Within 1 year, significant increases in uterus length from 73.4 mm to 88.2 mm, in uterus thickness from 33.9 mm to 43.5 mm and in endometrium height from 4.1 mm to 6.7 mm were observed (median values). There was an increase in both the number (from 2.2 to 3.5) and the size of the myomatas (29.4-35.0 mm diameter). A statistical analysis conducted by means of the Wilcoxon matched pairs signed-rank sum test showed P < 0.001. No significant change occurred in the size of the ovaries. Our study shows that hormone substitution may have an impact on uterus growth and that therefore vaginosonographical monitoring can be recommended.

Dehydroepiandrosterone↗

Histological assessment of ovaries and uterus of rats subjected to nandrolone decanoate treatment.

This study aimed to analyze the effects of nandrolone decanoate on the ovaries and uterus of adult females rats. This drug was administered intraperitoneally, at one, two and three doses of 3 mg nandrolone decanoate/kg of body weight, respectively, in the first, second and third week of treatment. The females of the control group received a physiological solution. The rats treated with nandrolone decanoate showed estral acyclicity and there was destruction of follicular units and an absence of corpus luteum in the ovaries. In the uterus, the drug promoted morphological alterations, characterized by vacuolated epithelium and endometrial stroma fibrosis. Ovary, uterus and pituitary weights were not affected by the steroid treatment. Nandrolone decanoate affects the sexual cycle and promotes histological alterations in the ovaries and uterus of adult female rats.

Anabolic Agents↗

[Contribution of three dimensional imaging techniques in the visualization of the fetal uterus].

The prenatal diagnosis of ambiguous genitalia requires a complete examination of the fetal anatomy to rule out any other congenital defects and the analysis of the fetal genotype. In addition any additional information on the anatomy of the internal genitalia, i.e. the presence or absence of fetal uterus, may be relevant to the prenatal evaluation of female pseudo-hermaphroditism. The recent development of 3D and 4D ultrasound technology may be relevant to a more clearly identification of the fetal uterus. Volume contrast imaging (VCI) provides high contrast images by the realization of thick slices of the region of interest (ROI) thereby providing a clearer picture of the fetal uterus. The multiplanar mode may also facilitate the differentiation of the fetal uterus from the other intra-pelvic organs by offering images in three perpendicular planes. Finally, the tomographic ultrasound imaging (TUI) mode offers the ability to display on a single panel numerous 2-dimensional sections, as obtained using computed tomography imaging.

Disorders of Sex Development↗

Fas ligand in the uterus of the non-pregnant mouse induces apoptosis of CD4+ T cells.

The role of FasL in the reproductive tissues of the non-pregnant mouse may be the induction of apoptosis of activated T cells for the prevention of unwanted inflammatory responses secondary to infection. This study was undertaken to identify cell and tissue types that typically express FasL in the uterus and oviduct of the mouse and to establish whether FasL on the surface of these cells was able to induce T cell apoptosis. FasL in the mouse uterus and oviduct was demonstrated using three independent methods: RT-PCR, Western blot and immunocytochemistry. The protein was present in the epithelial and mesenchymal cells of the uterus and showed a granular pattern in the apical epithelial portion. Although this suggests the presence of vesicles, surface expression was also detected by flow cytometry of isolated uterine cells. Exogenous administration of estradiol and progesterone had no significant effect on the expression and localization of FasL. The ability of uterine cells to induce FasL-dependent apoptosis of activated CD4+T cells was examined by incubation of phytohemagglutinin-treated T cells with cultured uterine cells. TUNEL and flow cytometric analyses showed that CD4+T cells experienced apoptosis after 5h of co-incubation. Neutralizing antibodies inhibited apoptosis demonstrating that a biologically active FasL is present in the reproductive tissues of the mouse. Results indicate that FasL is a biologically active molecule present in epithelial and mesenchymal cells of the uterus and the oviduct of the non-pregnant mouse that might restrain local immune response by induction of apoptosis of CD4+T lymphocytes.

Animals↗

Effect of phytoestrogens on gene expression of carbonic anhydrase II in rat uterus and liver.

The aim of this study was to characterize carbonic anhydrase II (CA2), as novel estrogen responsive gene, towards its usefulness to elucidate the molecular mechanisms of phytoestrogen action. Effects of estradiol-17beta (E2), and the phytoestrogens genistein (Gen), daidzein (Dai), as well as 8-prenylnaringenin (8PN) on CA2 mRNA expression were investigated in vivo in the uterus and liver of Wistar rats, and in vitro in Fe33 hepatoma cells. Relative amounts of mRNA levels of CA2 were measured by real-time RT-PCR. In vivo CA2 expression in uterus and liver is down-regulated by estrogen in time dependent manner with the most pronounced effect detectable 72 h after treatment. Treatment with Gen results in a slight down-regulation of CA2 expression in the uterus. In liver a response to Gen is detectable only after 7 h, where the expression of the gene is down-regulated to 60%. Treatment with Dai and 8PN for 72 h results in a slight down-regulation of CA2 in both tissues. In contrast in Fe 33 cells CA2 gene expression was up-regulated in response to the treatment with E2 for 7 h. In summary, we could demonstrate that the modulation of CA2 gene expression following treatment with E2 and Gen in rat uterus is comparable to the uterotrophic response of these substances, but with an inverted pattern. Remarkably, of all phytoestrogens 8PN exhibited the strongest uterotrophic response but only induced a very faint decrease of CA2 expression. In addition, we provide the first pieces of evidence that 8PN, like Gen and Dai, cannot be considered as a pure agonist. In conclusion, CA2 shows estrogen sensitivity not only in both tissues studied, but also in many others. Further, it exhibits a differential sensitivity thereby being capable to discriminate between different molecular qualities of phytoestrogens, like demonstrated for Gen and 8PN.

Animals↗

Tocolytic and toxic activity of papaya seed extract on isolated rat uterus.

Carica papaya L. seeds extracted with 80% ethanol (EEPS) caused concentration-dependent tocolysis of uterine strips isolated from gravid and non-gravid rats. Prostaglandin F2alpha and oxytocin-induced contractions of the isolated rat uterus were also inhibited in a concentration-dependent fashion by EEPS. Recoveries of the uterine activity after EEPS-induced uterine quiescence were very weak. Higher concentration of EEPS caused prompt uterine quiescence, which was also significantly irreversible. Pre-incubation of the rat uterus in Ringer Locke solution containing 10 mg/ml of EEPS for 1 hour prior to suspension in tissue baths led to significant depression of the spontaneous and KCl (60 mM)-induced uterine contractions relative to the solvent control (P<0.05). Cross sections of EEPS-pretreated non-gravid rat uterus (stained with hematoxyline and eosin) examined under light microscope revealed degeneration of the endometrium and myometrium with obvious cytoplasmic vacuolation indicating that EEPS could have direct toxic effect on the uterine tissues. Previous workers have reported benzyl isothiocyanate (BITC) as the main bioactive and anthelmintic compound in different extracts of papaya seeds. Using electron impact ionization methods, the presence of BITC in EEPS was also shown in this study. Mass spectra of both EEPS and standard BITC showed a base peak of benzyl/tropylium ion at m/z 91 (indicative of an aromatic compound) and the molecular ion peak of BITC (m/z 149). Our earlier studies have demonstrated BITC-induced functional and morphological derangement of isolated uterus. We thus conclude that at high concentration, EEPS is capable of causing irreversible uterine tocolysis probably due to the damaging effect of BITC (its chief phytochemical) on the myometrium.

Animals↗

Estrogen and phytoestrogen regulate the mRNA expression of adrenomedullin and adrenomedullin receptor components in the rat uterus.

The serum estrogen surge in the uterus triggers precisely-timed physiological and biochemical responses required establishing and maintaining pregnancy. Previous reports have shown that consumption of phytoestrogen-containing plants may disrupt the precise control of pregnancy. To evaluate the effects of phytoestrogens in the uterus, we screened for estradiol (E2)-inducible genes in immature rat uteri. We identified the gene for receptor-activity-modifying protein 2 (Ramp2), known to be a component of the adrenomedullin (ADM) receptor, as responsive to both E2 and the phytoestrogen coumestrol (Cou). We further examined the expression of ADM and ADM signaling components Ramp2, Ramp3, and CRLR in the immature rat uterus and found that both E2 and Cou regulated these genes expression. In addition, treatment with ADM increased uterine weight and edema similar to that observed after Cou treatment. Our findings indicated that the phytoestrogen caused the abnormal induction of vasoactive factors in the uterus.

Adrenomedullin↗

Tissue-specific modulation of cyclooxygenase-2 (Cox-2) expression in the uterus and the v. cava by estrogens and phytoestrogens.

Cyclooxygenase 2 (Cox-2), an enzyme involved in prostaglandin production, is a key player in the development of pathologic changes, such as colorectal cancer, arteriosclerosis and thrombosis. In this study, we investigated the effects of estrogens, selective estrogen receptor modulators (SERMs), pure antiestrogens and phytoestrogens on the tissue-specific expression of Cox-2 in the uterus and the v. cava of ovariectomized female rats. Cox-2 expression could be detected in the uterine epithelium and in the endothelium of the v. cava. Cox-2 expression was time-dependently stimulated after administration of 17beta estradiol (E2) in the uterus. In the v. cava, E2 treatment resulted in a stimulated expression of the progesterone receptor (PR), a gene known to be regulated by E2, whereas Cox-2 was simultaneously down-regulated. Administration of the pure antiestrogen faslodex (Fas) had no effect on Cox-2 expression. In contrast, administration of tamoxifen (Tam) resulted in a decrease of Cox-2 expression in the v. cava but does not stimulate Cox-2 expression in the uterus. Interestingly, the same expression pattern of Cox-2 could be detected after dose-dependent administration of genistein (Gen). Here, down-regulation of Cox-2 could already be detected after administration of merely 0.5 mg/(kgBW) Gen, a dose where no effects on uterine weight were observed. In summary, our results demonstrate a reverse tissue-specific regulation of Cox-2 expression by estrogens in the v. cava and uterus indicating the existence of complex molecular mechanisms which have to be characterized in future studies. Remarkably, Tam and the phytoestrogen Gen, both share the ability to decrease the expression of Cox-2 in the v. cava without effecting its uterine expression. These observations may be of great importance with respect to potential beneficial or adverse effects of estrogens, SERMs and phytoestrogens on the cardiovascular tissue.

Animals↗

The expression of Hedgehog genes (Ihh, Dhh) and Hedgehog target genes (Ptc1, Gli1, Coup-TfII) is affected by estrogenic stimuli in the uterus of immature female rats.

The objective of this study was to investigate the effects of estrogen receptor (ER) agonists and an ER antagonist on the expression of Hedgehog genes (Indian hedgehog: Ihh; Desert hedgehog: Dhh) and Hedgehog target genes (Patched 1: Ptc1; glioma-associated oncogene homolog 1: Gli1; chicken ovalbumin upstream promoter transcription factor II: Coup-TfII) in the rat uterus. Immature female rats were administered once with 17alpha-ethynyl estradiol (EE, an ER agonist), propyl pyrazole triole (PPT, an ERalpha-selective agonist), diarylpropionitrile (DPN, an ERbeta-selective agonist), or ICI 182,780 (an ER antagonist). Expression of mRNA for Ihh, Dhh, and Ptc1 was dose-dependently downregulated by EE in the uterus of immature rats, mediated by ER as confirmed by coadministration of ICI 182,780. The mRNA expression levels of Ptc1, Gli1, and Coup-TfII were simultaneously downregulated during the period in which the mRNA expression levels of Ihh and Dhh were downregulated in the uterus after administration of EE. PPT downregulated the transcription of Ihh, Dhh, Ptc1, Gli1, and Coup-TfII, indicating that expression of these genes was regulated by the ERalpha-dependent pathway. DPN also downregulated the transcription of Ihh and Dhh, although the effect was weaker than that of PPT, indicating that the regulation of uterine Ihh and Dhh transcription was also affected by the ERbeta-dependent pathway. These results suggest that the expression of Hedgehog genes (Ihh, Dhh) and Hedgehog target genes (Ptc1, Gli1, Coup-TfII) is affected by estrogenic stimuli in the uterus of immature female rats.

Animals↗