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Expression of corticotrophin-releasing hormone in the mouse uterus: participation in embryo implantation.

The detection of corticotropin-releasing hormone (CRH) in the pregnant and non-pregnant uterus has driven research to determine the role of this 41 amino acid neuropeptide in the female reproductive system. As concentrations of CRH mRNA and its peptide product are greater in the implantation sites of the early pregnant uterus compared with the regions between implantation sites, CRH has been hypothesised to participate in blastocyst implantation. Using the mouse system as an experimental model, we studied the distribution of CRH in the uterus during the oestrus cycle and early gestational period, and now provide evidence for its involvement in embryo implantation using cell culture techniques. The percentage of CRH-positive uterine cells and the amount of CRH released during anoestrus, pro-oestrus and oestrus were determined by immunofluorescence and ELISA experiments respectively. The highest number of intracellularly CRH-positive cells was obtained during pro-oestrus, whereas the highest CRH concentration in uterine cell culture supernatants was detected during anoestrus. At early stages of gestation, CRH was detected in the endometrium on days 2, 3 and 4 of pregnancy and in the myometrium on days 3 and 4, whereas it was undetectable on day 5. The functional role of CRH during early gestation was evaluated by administering anti-CRH antibody to mice from day 3 to day 8 of pregnancy. This treatment resulted in implantation failure in 60% of the cases, in which implantation sites, although clearly present in the uterus, had failed to host an embryo. These results provide direct evidence about the involvement of CRH in murine embryo implantation and are in agreement with hypotheses postulated in humans.

Animals↗

Localization and estrogenic regulation of androgen receptor mRNA expression in the mouse uterus and vagina.

Androgen receptors (AR) are highly expressed in female reproductive organs. In order to define the possible involvement of estrogens in the regulation of AR expression in the uterus and vagina, we have studied the effect of short-term administration of 17beta-estradiol (E2) to ovariectomized adult mice on AR mRNA levels. Seven days after ovariectomy, the mice received a single injection of E2 (0.05 microg/mouse) 3, 12 or 24 h before they were killed. The levels of AR mRNA were measured in the different uterine and vaginal compartments using quantitative in situ hybridization. In the uterus, AR mRNA was expressed in the luminal and glandular epithelial cells, stromal cells and smooth muscle cells. In the vagina, AR mRNA was localized in both epithelial and stromal cells. In the uterus after ovariectomy, AR mRNA levels were decreased by 18% in the epithelial cells, 23% in the stromal cells and 50% in the myometrial cells. AR mRNA levels were completely restored as early as 3 h after E2 administration in the epithelium and stroma, and at the 12-h time-interval in the myometrium. In the vaginal epithelium, ovariectomy induced a 70% decrease in AR mRNA expression. No effect could be detected 3 h after E2 administration, while at the longest time-intervals (12 and 24 h) there was an increase in mRNA levels corresponding to 70% of the levels observed in intact animals. In the vaginal stroma, ovariectomy was responsible for a 55% decrease in mRNA levels. While no significant changes were observed at the 3-h time-interval, a complete restoration of AR mRNA levels in stromal cells could be recorded at the longest time-intervals after E2 administration. The data obtained indicated that, in adult mice, estrogens exert a positive regulation of AR mRNA expression in the different compartments of both the uterus and the vagina.

Animals↗

Effects of peroxisome proliferator activated receptors-alpha and -gamma agonists on estradiol-induced proliferation and hyperplasia formation in the mouse uterus.

It is suggested that the action of peroxisome proliferator-activated receptors (PPARs) cross-talks with estrogen signaling in the uterus. However, it is not known how PPAR agonists affect estrogen-dependent processes in the uterus, especially proliferation and morphogenetic changes. The effects of agonists of PPAR-alpha and -gamma on proliferative and morphogenetic reactions in the uterus under short- and long-term estrogen treatments were therefore examined. Ovariectomized mice were treated with estradiol dipropionate (4 micro g/100 g, s.c., once a week) or vehicle and rosiglitazone (PPAR-gamma agonist) or fenofibrate (PPAR-alpha agonist) or with no additional treatment for 2 days or for 30 days. Treatment with estradiol and PPAR agonists for 2 days did not affect uterine mass. In mice treated with estradiol and rosiglitazone for 2 days, proliferation was enhanced and levels of estrogen receptors-alpha and beta-catenin were decreased in all uterine tissues. Treatment with estradiol and fenofibrate for 2 days had the opposite effects on the parameters tested. In animals treated with estradiol and rosiglitazone for 30 days, uterine mass was increased, abnormal uterine glands and atypical endometrial hyperplasia were found more often and levels of estrogen receptors-alpha and beta-catenin were decreased. In animals treated with estradiol and fenofibrate for 30 days, uterine mass was decreased, most of the uterine glands had a normal structure, no cases of atypical hyperplasia were diagnosed, proliferative activity was declined and the levels of estrogen receptors-alpha and beta-catenin were markedly higher. Treatment with rosiglitazone or fenofibrate did not affect the serum estradiol level in the mice which received estradiol together with PPAR agonists for 30 days. Thus, rosiglitazone exerted the proliferative and morphogenetic effects of estradiol, but fenofibrate had the opposite effect. The actions of rosiglitazone and fenofibrate are associated with changes in the expression of estrogen receptors-alpha and beta-catenin in the uterus.

Animals↗

The estrogen receptor alpha sigma3 mRNA splicing variant is differentially regulated by estrogen and progesterone in the rat uterus.

The gene for estrogen receptor alpha (ER alpha) has been shown to be under complex hormonal control and its activity can be regulated by mRNA alternative splicing. Here we examined the regulation of ER alpha transcription and translation in the rat uterus by ovarian steroid hormones. We examined whether expression of ER alpha mRNA splice isoforms is hormonally regulated in ovariectomized (OVX) and cycling rats. Adult OVX female rats were treated daily with 17-beta estradiol (E2) (0.05 microg/rat or 5 microg/rat), progesterone (P4) (1 mg/rat) or a combination of both hormones for 4 days. Animals were killed 24 h after the last injection and uterine horns were removed. In order to determine whether ER alpha mRNA isoforms are differentially expressed under various physiological conditions, animals were evaluated at proestrus, estrus and diestrus. The ER alpha protein and mRNA were detected by immunohistochemistry and comparative RT-PCR analysis respectively. The presence of ER alpha mRNA isoforms was evaluated using a nested RT-PCR assay. In OVX control rats, ER alpha mRNA and protein levels were high, demonstrating a constitutive expression of the ER alpha gene in the uterus. When animals received P4 or the high dose of E2, a significant decrease in both ER alpha mRNA and protein was observed in the uterus. However, when rats were protein was treated with the low dose of E2, only the ER alpha down-regulated; no changes were observed in ER alpha mRNA expression. In addition to the full-length ER alpha mRNA, OVX control rat uteri expressed three shorter transcripts: sigma3, sigma4 and sigma3,4 (lacking exon 3, exon 4, or both 3 and 4 respectively). Surprisingly, when OVX animals were treated with P4, the low dose of E2 or a combination of both steroids, expression of the sigma3 isoform was completely abolished. During the estrous cycle, all ER alpha mRNA splicing variants were detected at proestrus and estrus. However, in diestrus, significant low levels of the sigma3 isoform were observed. In summary, our results suggest a dose-dependent relationship between E2 concentrations and the level of control in the ER alpha transcription-translation cascade. Moreover, the alternative splicing of the ER alpha primary transcript is influenced by the hormonal milieu, suggesting that these events could affect the estrogen responsiveness of the rat uterus during the estrous cycle.

Alternative Splicing↗

Effects of histone deacetylase inhibitors on estradiol-induced proliferation and hyperplasia formation in the mouse uterus.

It is suggested that estrogen hormones recruit mechanisms controlling histone acetylation to bring about their effects in the uterus. However, it is not known how the level of histone acetylation affects estrogen-dependent processes in the uterus, especially proliferation and morphogenetic changes. Therefore, this study examined the effects of histone deacetylase blockers, trichostatin A and sodium butyrate, on proliferative and morphogenetic reactions in the uterus under long-term estrogen treatment. Ovari-ectomized mice were treated with estradiol dipropionate (4 microg per 100 g; s.c., once a week) or vehicle and trichostatin A (0.008 mg per 100 g; s.c., once a day) or sodium butyrate (1% in drinking water), or with no additional treatments for a month. In animals treated with estradiol and trichostatin A or sodium butyrate, uterine mass was increased, and abnormal uterine glands and atypical endometrial hyperplasia were found more often. Both histone deacetylase inhibitors produced an increase in the numbers of mitotic and bromodeoxyuridine-labelled cells in luminal and glandular epithelia, in stromal and myometrial cells. Levels of estrogen receptor-alpha and progesterone receptors in uterine epithelia, stromal and myometrial cells were decreased in mice treated with estradiol and trichostatin A or sodium butyrate. Expression of beta-catenin in luminal and glandular epithelia was attenuated in mice treated with estradiol with trichostatin A or sodium butyrate. Both histone deacetylase inhibitors have similar unilateral effects; however the action of trichostatin A was more expressed than that of sodium butyrate. Thus, histone deacetylase inhibitors exert proliferative and morphogenetic effects of estradiol. The effects of trichostatin A and sodium butyrate are associated with changes in expression of estrogen receptor-alpha, progesterone receptors and beta-catenin in the uterus.

Animals↗

Changes in action potential and beta-adrenergic effects of the circular muscle of postpartum rat uterus.

Spike potential dominated in the circular muscle of postpartum rat uterus during the period between 0 and 15 hr after the delivery of the first newborn. During the postpartum period ranging between 20 and 48 hr, the plateau potential was dominant. Application of 10(-9) M isoprenaline strongly depressed the contraction during early postpartum period (0-15 hr), and the depressant effect was much smaller thereafter. In vivo treatment of postparturient rats with estradiol-17 beta (50 micrograms) or progesterone (50 micrograms) for 2 days did not alter the postpartum change in action potential or the effect of isoprenaline. The postpartum changes in muscle properties mentioned above were prevented in the distended portion of uterus, when several fetuses and placentas were artificially kept inside the uterus for 2 days, while other fetuses were delivered out. The hormonal influences on the circular muscle of postpartum rat uterus were discussed in view of the above experimental findings.

Action Potentials↗

Procedures for measuring length of the pig uterus.

A method for measuring uterine length in pigs was standardized following a series of experiments to determine the effect of period of storage, temperature, presence of ligaments and the nature of the surrounding fluid on the length of the uterus. Weight of the uterus did not change during storage. Measurements in vivo were compared with those in vitro. The uterus shortened immediately after its removal, then gradually lengthened over 8 h. Whereas there was change in absolute measured length during the storage period, the relationship of length in vivo was significantly correlated to length in vitro (r = .85, P less than .001). Measurement at one time during storage was highly correlated with measurement at other times (r = .93 to .96). The coefficient of variation for length of uteri among gilts was 30%. Measurement of length of the uterus must be done under uniform conditions of storage to give meaningful results.

Animals↗

Twin pregnancy with a fetus in each half of a uterus didelphys.

A rare case of a twin pregnancy with a fetus in each half of a uterus didelphys (double uterus, double cervix and septate vagina) is reported. A longitudinal vaginal septum and two portios were detected during the first labor of this patient. During her second pregnancy ultrasonography was performed in the 16th week, and pregnancy was detected in each half of the double uterus. Both fetuses were of similar size and corresponded to the gestational age. A completely separated double uterus was confirmed by ultrasonography. In the 38th week a female and a male infant were delivered by cesarean section. The follow-up of this pregnancy and the management of the labor are reported.

Adult↗

The increasing incidence and declining mortality of ruptured uterus in Enugu.

A 10-year review of ruptured uterus at the University of Nigeria Teaching Hospital disclosed an incidence of 1 in 500 deliveries. The average incidence for booked patients alone was 1 in 1271 deliveries. The maternal and perinatal mortalities were 17% and 91% respectively, while the contributions of ruptured uterus to the overall hospital maternal and perinatal mortalities for the period under review were 4.1 and 2.6% respectively. In 1978, the average incidence of ruptured uterus in all patients delivered at the hospital increased from 1 in 787 in the 1st 5 years to 1 in 330 in the 2nd 5 years, while the maternal mortality from ruptured uterus decreased from 27% in the 1st 1/2 to 10% in the 2nd 1.2 of the study. The decline in maternal mortality in this environment is due to better availability of stored blood and management which included subtotal hysterectomy in many cases.

Africa↗

The distribution of collagenase in the rat uterus during postpartum involution. An immunohistochemical study.

A monospecific rabbit anti-rat uterus collagenase antibody has been prepared and used to study the distribution of the enzyme in the rat uterus during postpartum involution. Cryostat sections of rat uteri from 24 to 240 hours postpartum were stained by the indirect immunofluorescent method. Nonpregnant rat uterus revelaed positive staining in basement membranes, in endometrial stroma, in perimuscular and in vascular connective tissue. During postpartum involution of the uterus two types of changes in uterine collagenase were observed: (1) variations in the distrubiton of the enzyme, which became selectively localized in the epithelial basement membrane and in the wall of small blood vessels, and (2) variations in the overall intensity of fluorescent staining, which decreased immediately after delivery and slowly increased back to nonpregnant levels in 5 days. The significance of these findings is discussed in relation to the mechanisms of control of collagenase activity in vivo.

Animals↗

Progesterone binding in rabbit oviduct and uterus.

Progesterone binding of high affinity with a dissociation constant of 10(-9) M was identified in cytosol of rabbit oviduct and uterus. Macromolecules with sedimentation coefficients of 7-8 S and 4-5 S were present. Progesterone receptor concentration was two to fivefold lower in the oviduct when compared with the uterus. The receptor concentration declined steadily from 3 hr until 144 hr after mating in the uterus; however, the decline in oviductal receptor was not significant until the sixth day of pregnancy. Progesterone receptor concentration in rabbit oviduct and uterus in estrus and early pregnancy was greater than estradiol receptor levels.

Animals↗

Differential expression of VEGF isoforms and VEGF(164)-specific receptor neuropilin-1 in the mouse uterus suggests a role for VEGF(164) in vascular permeability and angiogenesis during implantation.

The mechanism(s) by which localized vascular permeability and angiogenesis occur at the sites of implantation is not clearly understood. Vascular endothelial growth factor (VEGF) is a key regulator of vasculogenesis during embryogenesis and angiogenesis in adult tissues. VEGF is also a vascular permeability factor. VEGF acts via two tyrosine kinase family receptors: VEGFR1 (Flt-1) and VEGFR2 (KDR/Flk-1). Recent evidence suggests that neuropilin-1 (NRP1), a receptor involved in neuronal cell guidance, is expressed in endothelial cells, binds to VEGF(165) and enhances the binding of VEGF(165) to VEGFR2. We examined the spatiotemporal expression of vegf isoforms, nrp1 and vegfr2 as well as their interactions in the periimplantation mouse uterus. We observed that vegf(164) is the predominant isoform in the mouse uterus. vegf(164) mRNA accumulation primarily occurred in epithelial cells on days 1 and 2 of pregnancy. On days 3 and 4, the subepithelial stroma in addition to epithelial cells exhibited accumulation of this mRNA. After the initial attachment reaction on day 5, luminal epithelial and stromal cells immediately surrounding the blastocyst exhibited distinct accumulation of vegf(164) mRNA. On days 6-8, the accumulation of this mRNA occurred in both mesometrial and antimesometrial decidual cells. These results suggest that VEGF(164) is available in mediating vascular changes and angiogenesis in the uterus during implantation and decidualization. This is consistent with coordinate expression of vegfr2, and nrp1, a VEGF(164)-specific receptor, in uterine endothelial cells. Their expression was low during the first 2 days of pregnancy followed by increases thereafter. With the initiation and progression of implantation (days 5-8), these genes were distinctly expressed in endothelial cells of the decidualizing stroma. Expression was more intense on days 6-8 at the mesometrial pole, the presumptive site of heightened angiogenesis and placentation. However, the expression was absent in the avascular primary decidual zone immediately surrounding the implanting embryo. Crosslinking experiments showed that (125)I-VEGF(165) binds to both NRP1 and VEGFR2 present in decidual endothelial cells. These results suggest that VEGF(164), NRP1 and VEGFR2 play a role in VEGF-induced vascular permeability and angiogenesis in the uterus required for implantation. genesis 26:213-224, 2000.

Animals↗

[Vaginal hysterectomy in non-prolapsed uterus].

AIM OF STUDY: To describe the results of vaginal removal of the non-prolapsed uterus in women conventionally operated on abdominally or with the assistance of laparoscopy. MATERIAL AND METHODS: At the departments of gynaecology, Herning Central Hospital and Holstebro Central Hospital, approximately one third of all hysterectomies are performed vaginally. All records of women, who had had a vaginal hysterectomy on a non-prolapsed uterus over a nineteen-month period, were reviewed retrospectively. Women, who had additional surgery, were excluded. RESULTS: One-hundred and thirteen patients entered. In one patient (1%) the vaginal hysterectomy was converted to an abdominal one. One fourth of the women had a uterus weighing more than 200 grams. The median operation time was 58 minutes; 73% bled less than 200 ml. Half of the women were discharged from hospital on the third postoperative day or earlier; 90% on the fifth postoperative day or earlier. During operation three accidental bladder lesions occurred, and four women needed an additional haemostatic operation. Postoperatively, 10 women (9%) suffered from a haematoma or an abscess in the vaginal vault. CONCLUSIONS: Vaginal hysterectomy on the non-prolapsed uterus is an operation that should be offered to a large group of women, who today are operated on abdominally or with laparoscopic assistance. The operation is quick and the patients are discharged after a few days. No advanced equipment is needed. Some women will, however, suffer from a haematoma in the vaginal vault.

Female↗

Immunoreactivity of iNOS in porcine uterus after infusions of Escherichia coli endotoxin.

The aim of this study was to investigate the distribution of inducible isoform of nitric oxide synthase (iNOS) in the porcine uterus after infusions of Escherichia coli endotoxin (lipopolysaccharide, LPS). In the group I (treated; n=6), 1 mg of LPS was infused into both the left and right uterine horn starting from the 4th to the 10th day of the estrous cycle, twice a day. In the group II (control; n=6), saline was infused into the uterus. The uterine horns were collected on the 14th day of the estrous cycle. Cryostat sections from the paraformaldehyde fixed tissues were stained immunohistochemically to estimate the distribution of iNOS. The luminal and glandular epithelium was stained more intensely for iNOS in the LPS-treated gilts than in the control animals. After LPS infusions, iNOS staining in vascular endothelial cells was also more intense than that observed in the controls. The present study has revealed that infusions of LPS into the porcine uterus result in an increase in the intensity of iNOS staining in some structures of this organ and supports our earlier data that NO can mediate an inflammatory effect of LPS in the uterus.

Animals↗

Metabolism of norethisterone and norethisterone derivatives in rat uterus, vagina, and aorta.

The 19-nor-progestogen norethisterone is used as a progestogen component in contraceptives and in continuous- and sequential combined hormone replacement therapy (HRT) in postmenopausal women. Metabolism of norethisterone in HRT target tissues may play a role in its biological response. The aim of this study was to investigate which steroid-metabolizing enzymes are present in rat uterus, vagina, and aorta, three HRT target tissues. Next, the ability of the tissues to metabolize norethisterone was assessed. Furthermore, to investigate the effect of substituents at the 7- and 11-position, the metabolism of Org OM38 (7alpha-methyl-norethisterone), Org 4060 (11beta-ethyl-norethisterone), and Org 34694 (7alpha-methyl,11-ethylidene-norethisterone) was studied. Using radiolabeled progesterone, the presence of 20alpha-hydroxysteroid dehydrogenase, 5alpha-reductase, and 3alpha-hydroxysteroid dehydrogenase activity could be demonstrated in uterus, vagina, and to a lesser extent in aorta. The combined action of the latter two enzyme activities resulted in 3alpha-OH,5alpha-H-norethisterone as the major metabolite of radiolabeled norethisterone in uterus (26.9%), vagina (37.1%), and aorta (1.4%). The norethisterone derivatives, however, were metabolized to a much lesser extent (1.0-7.6%). No formation of 5alpha-reduced forms of Org 4060, Org OM38, or Org 34694 was found, while formation of minor amounts of 3alpha-OH-Org 4060 and 3alpha-OH-Org OM38 could be demonstrated in both uterus, vagina, and aorta. These findings confirm the role of 5alpha-reductase as a rate-limiting step in the metabolism of norethisterone derivatives and show important inhibitory effects of substituents at the 7alpha- and 11-position of the steroid skeleton on 5alpha-reduction.

Animals↗

Replacement of the uterus by malignant mesothelioma of the peritoneum: a case report.

Malignant mesothelioma of the peritoneum is a rare tumor that must be distinguished from the more common primary peritoneal serous epithelial neoplasms. We report a case of a 66-year-old female presenting with weight loss, anemia, and a large pelvic mass on ultrasound. At laparotomy a large (9 x 6.5 x 3.5 cm) mass in the anatomical position of the uterus was found in the minor pelvis. The uterus, tubes and ovaries could not be identified. Palpable paraaortic lymph nodes and liver nodules were found. Immunohistochemically the tumor cells were positive for cytokeratin, epithelial membrane antigen and vimentin-CEA, but S-100 protein and Leu-M1 were negative. Remnants of the uterine corpus, fallopian tubes and ovaries could not be identified. The overall features were best regarded as malignant peritoneal mesothelioma. There have been reports of mesotheliomas involving the uterus. However, this is the first reported case of mesothelioma causing total replacement of the uterus.

Aged↗

[Infantile and hypoplastic uterus: a contribution to overcome the confusion of terms (author's transl)].

The confusion of terms between the radiological literature and the gynaecological and anatomical literature regarding the definitions of the infantile and the hypoplastic uterus are described. The history of this confusion is traced. The two types of uterus were clearly defined by K. Menge and K.v. Oettingen in 1930. The infantile uterus shows an abnormal relationship between the length of the cervix and the length of the uterine cavity of 1:1 or 2:1, whereas the hypoplastic uterus shows a normal relationship between the length of the cervix and the length of the uterine cavity of approximately 1:2.

Congenital Abnormalities↗

[Pharmacological models to investigate the role of the adrenergic system on post-partum rat uterus].

The adrenergic system plays a major role in the regulation of the pregnant uterine contractility. Our aim was to develop an experimental animal model to study the role of the alpha 1A-adrenergic receptor (AR) in uterine motor activity by antisense oligonucleotides (AONs). AONs were injected with DOTAP and pluronic gel into the uterine lumen of post-partum rats 2-3 hours after delivery. The decrease of the alpha 1A-AR density by AON was demonstrated by RT-PCR method, Western blot analysis and radioligand binding assay on rat uterus preparations 24 h after delivery. The changes in the contractility of the uterus were measured on isolated rat uterine tissue by electric field stimulation (EFS). The EFS investigation demonstrated that the effect of the specific alpha 1A-blocker 5-methylurapidil and WB4101 was significantly decreased in the AON-treated rat uterus as compared to the control group but the effect of the beta-mimetic terbutalin and alpha 1D-antagonist BMY7378 was unchanged. Our result suggest that the alpha 1A-ARs play a very important role in the regulation of uterine contractility, and may serve as the basis for a subsequent new group of tocolytics (uterus selective alpha 1-antagonists), which may lead to more selective therapy than currently used beta-mimetics.

Adrenergic alpha-Antagonists↗