Paracrine regulation of epithelial progesterone receptor by estradiol in the mouse female reproductive tract

Regulation of progesterone receptor (PR) by estradiol-17beta (E(2)) in mouse uterine and vaginal epithelia was studied. In ovariectomized mice, PR expression was low in both vaginal stroma and epithelium, but high in uterine epithelium. E(2) induced PR in vaginal epithelium and stroma, but down-regulated PR in uterine epithelium. Analysis of estrogen receptor alpha (ERalpha) knockout (ERKO) mice showed that ERalpha is essential for E(2)-induced PR expression in both vaginal epithelium and stroma, and for E(2)-induced down-regulation, but not constitutive expression of PR in uterine epithelium. Regulation of PR by E(2) was studied in vaginal and uterine tissue recombinants made with epithelium and stroma from wild-type and ERKO mice. In the vaginal tissue recombinants, PR was induced by E(2) only in wild-type epithelium and/or stroma. Hence, in vagina, E(2) induces PR directly via ERalpha within the tissue. Conversely, E(2) down-regulated epithelial PR only in uterine tissue recombinants constructed with wild-type stroma. Therefore, down-regulation of uterine epithelial PR by E(2) requires stromal, but not epithelial, ERalpha. In vitro, isolated uterine epithelial cells retained a high PR level with or without E(2), which is consistent with an indirect regulation of uterine epithelial PR in vivo. Thus, E(2) down-regulates PR in uterine epithelium through paracrine mechanisms mediated by stromal ERalpha.     

Effects of estradiol and progesterone on scent-marking behavior of female rats

The effect of estradiol and progesterone on scent-marking behavior of the female rat is reported. Estradiol followed by progesterone injection to ovariectomised rats results in an increase in marking rates. This suggests an endocrine base to the changes in scent-marking behavior that are known to occur with the rat estrous cycle.     

Differential suppression of progesterone receptors by progesterone in the reproductive tract of female macaques

Ovariectomized cynomolgus macaques were treated with implants of estradiol (E2) for 14 days. Some animals then received an additional implant of progesterone (P) for 7-14 more days. After treatment with either E2 alone or with E2 plus P we removed the reproductive tracts and measured nuclear and cytosolic P receptors by exchange assay. In addition we used steroid radioimmunoassays(RIA) to measure levels of E2 and P in parallel aliquots of the nuclear and cytosolic fractions. P treatment reduced the concentrations of E2 in nuclear and cytosolic fractions in the cervix, endometrium, myometrium and oviduct compared to the amounts present after 14 days of E2; these data are consistent with many reports that P treatment significantly lowers the amount of nuclear and cytosolic estrogen receptors in all of these tissues. In the oviduct, myometrium and cervix both cytosolic and nuclear P receptor levels were lowered during P action. In the endometrium, however, P treatment reduced the amount of P receptor only in the cytosolic but not the nuclear fraction. RIA determinations of the amount of P retained in nuclear fractions of the P-treated animals indicate that P levels were significantly elevated only in the nuclei obtained from endometrium. This specific increase in the retention of P by endometrial nuclei during P action is consistent with the specific retention of P receptor by endometrial nuclei. These results lead to the unexpected conclusion that the stimulatory effects of P as expressed in the maintenance of the progestational state in the primate endometrium may require higher levels of occupied nuclear P receptor than do the suppressive effects of P as expressed in oviductal atrophy, diminished cervical secretion and myometrial quieting.     

Effects of estradiol and progesterone on the reproductive tract and on uterine sex steroid receptors in female lambs

The effects of estradiol-17 beta (E(2)) and progesterone (P) on the reproductive tract and on uterine estrogen receptors and P receptors were studied in 2-mo-old female lambs (n = 11). On Days 0, 1 and 2, E(2) (1 ug/kg, Group E, n = 4), P (0.3 mg/kg, Group P, n = 4) or corn oil (control) vehicle (Group C, n = 3) were administered, and in Day 3 all lambs were slaughtered. Group E (n = 12) had E(2) serum concentrations (mean +/- SEM) of 43.8 +/- 2.2 pmol L , similar to that of the follicular phase; while P concentrations in Group P (n = 12) were similar (2.8 +/- 0.18 nmol L ) to those of the luteal phase of the ewe estrous cycle. The E(2) treatment increased the reproductive tract weight, while P treatment increased only the uterine weight. Both E(2) and P receptors from upper and middle uterine zones (including the myometrium, endometrium and caruncles) were determined by binding assays with tritiated hormones, dextran-charcoal separation and inverse Scatchard analysis. Both the E(2) and P treatments decreased E(2) and P receptor concentrations in upper and middle zones, although the upper zone had higher receptor concentrations than the middle zone (P < 0.01). E(2) receptor concentrations in the upper zone (mean +/- SEM, fmol mg prot) were 1236 +/- 34, 667 +/- 80 and 444 +/- 103 for Groups C, P and E, respectively. The P receptor concentrations were 2434 +/- 135, 1273 +/- 102 and 1536 +/- 213 for the same groups. The high uterine P receptor concentrations allowed P action without prior estrogen priming of female lambs. The present results suggest that E(2) and P might down-regulate their own and each other's receptors during development. The biological responses induced by E(2) and P, as measured by the reproductive tract weight, demonstrated that at an early stage of development uterine receptors are physiologically active.     

Effects of estradiol and progesterone on the sensitivity to pain and on morphine-induced antinociception in female rats

Long-term ovariectomized (OVX) rats were exposed to 2- or 14-day replacement with pellets made of cholesterol (CHOL), estradiol (E2), progesterone (P4), or a combination of E2 and P4. Following the treatment with steroids the antinociceptive effect of morphine (5 mg/kg,sc) was measured by a hot-plate method. Pellets of E2 (0.5 and 5%) caused dose- and time-dependent reductions of morphine-induced antinociception as compared with OVX rats treated with CHOL pellets. Moreover, OVX rats pretreated with E2 pellets had decreased basic sensitivity to nociceptive stimulus (hyperalgesia). Treatment for 2 and 14 days with 75% P4 pellets produced significant reduction of MOR antinociception. The low dose of P4 (10% pellet) did not change the effect of MOR on Day 2 but significantly increased the antinociceptive effect of MOR on Day 14. Replacement of OVX rats with one 0.5% E2 pellet plus one 10% P4 pellet resulted in marked inhibition of the antinociceptive effect of MOR on Day 2 as well as on Day 14. Central injection 30 min before MOR of either LHRH antagonist or the antiserum against LHRH into OVX rats pretreated for 14 days with both steroids had no effect on the degree of the antinociception. The results suggest that the effects which ovarian steroids exert on opioid systems vary according to the dose, the duration of treatment, and the type of steroid administered.     

Activation of AMPA receptors inhibits prolactin and estradiol secretion and delays the onset of puberty in female rats

Previous experiments have evidenced the neuroendocrine role of AMPA receptors. Present studies were carried out to obtain information on the role of these receptors in the control of the onset of puberty. To this end, female rats were i.c.v. injected with vehicle or AMPA (agonist of AMPA receptors: 0.1 or 0.5 nmol/day) between 26 and 30 days (Experiment 1), or 30 and 34 days (Experiment 2) of age. Serum concentrations of PRL, LH and estradiol were measured before drug administration, 10 min after the last injection, at vaginal opening (VO) and at first estrus (FE) presentation. In both experiments, AMPA administration inhibited PRL and estradiol secretion without affecting LH release. When AMPA was administered between 26 and 30 days a significant delay in the day of vaginal opening was observed. These results confirmed the inhibitory effect of AMPA on PRL secretion and suggests a role of AMPA receptors in the control of puberty onset.     

Preference for an estrous female over a non-estrous female evinced by female rats requires dihydrotestosterone plus estradiol

The effects were studied of long-term treatment with testosterone metabolites (dihydrotestosterone, DHT, and estradiol, E2, in sc Silastic implants) on preference behavior of ovariectomized female rats for an estrous female over a non-estrous female. For measuring this behavior a residential plus-maze was used which harbored two ovariectomized “stimulus” females on the top of peripheral boxes, one of which was made estrus by injection of estradiol benzoate and progesterone. When both steroids (DHT plus E2) were circulating simultaneously they evoked preference for an estrous female, while neither steroid by itself sufficed. In earlier work with adult male rats castrated on the day of birth, E2 was effective in the absence of DHT. This sex difference, therefore, seems to have arisen before birth. Further, administration of DHT alone caused a profound lack of interest in both “stimulus” females, which cannot be fully explained by the reduced locomotor activity which has been found to be induced by DHT in earlier Studies.     

Chronic treatment with estradiol restores depressive-like behaviors in female Wistar rats treated neonatally with clomipramine

Neonatal administration of clomipramine (CMI) induces diverse behavioral and neurochemical alterations in adult male rats that resemble major depression disorder. However, the possible behavioral alterations in adult female rats subjected to neonatal treatment with clomipramine are unknown. Therefore, the aim of this study was to explore the effect of neonatal treatment with CMI on adult female rats in relation to locomotion and behavioral despair during the estrus cycle. Also evaluated was the effect of chronic treatment with E2 on these female CMI rats. We found no effects on spontaneous locomotor activity due to neonatal treatment with CMI, or after 21 days of E2 administration. In the FST, neonatal treatment with CMI increased immobility and decreased active swimming and climbing behaviors. Influence of the ovarian cycle was detected only in relation to climbing behavior, as the rats in the MD phase displayed less climbing activity. Chronic E2 administration decreased immobility but increased only swimming in CMI rats. These results suggest that neonatal treatment with CMI induces despair-like behavior in female rats, but that chronic E2 administration generates antidepressant-like behavior by decreasing immobility and increasing swimming, perhaps through interaction with the serotonergic system.     

Effects of progesterone and estradiol benzoate on glutathione dependent antioxidant enzyme activities in the brain of female rats.

The activities of glutathione dependent antioxidant enzymes were measured in subcellular fractions of whole brain homogenates prepared from ovariectomized (OVX) female rats, untreated or treated 2 h or 24 h prior to sacrifice with a single dose of 2 mg progesterone (P) or 5 micrograms estradiol benzoate (EB). Glutathione peroxidase (GSH-Px) activity was not changed following systemic administration of EB, but P increased GSH-Px in the brain of OVX rats 24 h after the treatment. The activity of glutathione reductase (GR) was suppressed by EB short time, only 2 h following treatment, whereas P increased the enzyme activity 24 h after treatment. On the other hand, the activities of catalase (CAT) and glutathione-S-transferase (GST) were not changed following systemic administration of EB or P. The present work was carried out to study the involvement of ovarian steroids, especially P, in the control of GSH-Px and GR activities, and our results suggest that oxidative stress in the brain of female rats may be modulated by the level of progesterone.     

Does estradiol treatment normalize the hypothalamic-pituitary-adrenal axis in streptozotocin-induced ovariectomized diabetic female rats?

We recently found circulating corticosterone (CS) levels to be significantly lower in diabetic female rats as compared with proestrous control animals. This reduction in CS was correlated with the hypoestrogenic state of the diabetic female. It was the purpose of this study to evaluate basal and corticotropin releasing hormone (CRH)-stimulated CS secretion in ovariectomized (OVX) control (C) and streptozotocin-induced diabetic (D) rats given blank, 5 mcg and 20 mcg estradiol (E2) implants to determine if adrenal CS secretion in the diabetic is normalized by E2 treatment. After 3 weeks of diabetes, pituitary-adrenal function was assessed in rats from each group with a CRH stimulation test. The remaining rats were sacrificed for determination of CS, E2, testosterone and fructosamine in serum. Suppressed CS secretion in OVX female diabetic rats was partially restored with E2 therapy. Basal CS levels were significantly higher in 20 mcg E2 treated C and D rats compared with OVX rats. However, C rats had significantly higher basal CS compared with D rats in similarly E2 treated groups. The CS response to CRH stimulation was not different between OVX female diabetic and control rats. Estrogen enhanced the CS response to CRH stimulation in control animals but not in diabetic animals suggesting altered estrogen action at the pituitary level in diabetic animals.     

The effect of progesterone and progesterone + estradiol on the morphology of the pineal gland in immature female pigs

Pineal gland of the immature female pigs treated with progesterone and progesterone and estradiol simultaneously were investigated with light and electron microscopy. Both of the applied hormones influenced the structure of the pineal glands. Inhibitory effect of progesterone on the relative volume of mitochondria, granular vesicles and dense bodies of type one was observed. Administration of progesterone and estradiol simultaneously caused more complicated influence statistically significant increase of pinealocyte nuclei volume and at the same time significant decrease of mitochondria and granular vesicles.     

Intra-striatal estradiol in female rats impairs response learning within two hours of treatment

Estradiol treatment administered systemically or directly to the dorsolateral striatum across two days impairs performance on a response task in which rats learn to make a specific body turn to locate food on a maze. Estradiol can act through both slow and rapid signaling pathways to regulate learning impairments, however it is impossible to dissociate the slow from the rapid contributions of estradiol following long exposures. To assess the rapid effects of estradiol on striatum-sensitive learning, we trained rats on a response learning task after either relatively short or long treatments of estradiol infused directly into the striatum. Three-month-old female rats were ovariectomized 21 days before training and received guide cannulae implanted bilaterally into the dorsolateral striatum. For short duration treatments, rats were given bilateral infusions (0.5 μl) of 17β-estradiol-sulfate (0, 5, 50, or 500 nM in aCSF-vehicle) either 2 h or 15 min prior to training. For long duration treatments, rats received a series of estradiol infusions (500 nM) at 48, 24, and 2 h prior to training. Replicating previous findings (Zurkovsky et al., 2007), intra-striatal estradiol treatments given for two days prior to training impaired response learning. Estradiol-induced impairments in performance were also demonstrated 2 h, but not 15 min, after single infusions. Thus, estradiol acts within hours of exposure in the striatum, a structure lacking classical estrogen receptors, to impair response learning.     

Intra-striatal estradiol in female rats impairs response learning within two hours of treatment

Estradiol treatment administered systemically or directly to the dorsolateral striatum across two days impairs performance on a response task in which rats learn to make a specific body turn to locate food on a maze. Estradiol can act through both slow and rapid signaling pathways to regulate learning impairments, however it is impossible to dissociate the slow from the rapid contributions of estradiol following long exposures. To assess the rapid effects of estradiol on striatum-sensitive learning, we trained rats on a response learning task after either relatively short or long treatments of estradiol infused directly into the striatum. Three-month-old female rats were ovariectomized 21 days before training and received guide cannulae implanted bilaterally into the dorsolateral striatum. For short duration treatments, rats were given bilateral infusions (0.5 μl) of 17β-estradiol-sulfate (0, 5, 50, or 500 nM in aCSF-vehicle) either 2 h or 15 min prior to training. For long duration treatments, rats received a series of estradiol infusions (500 nM) at 48, 24, and 2 h prior to training. Replicating previous findings (Zurkovsky et al., 2007), intra-striatal estradiol treatments given for two days prior to training impaired response learning. Estradiol-induced impairments in performance were also demonstrated 2 h, but not 15 min, after single infusions. Thus, estradiol acts within hours of exposure in the striatum, a structure lacking classical estrogen receptors, to impair response learning.     

Estradiol plus progesterone treatment and precopulatory behavior in prepubertally ovariectomized female rats: dose-response relationships

The copulatory and precopulatory behavioral repertoire expressed in terms of the intensity scale of sexual responsiveness was investigated in female rats ovariectomized on the 30th day after birth and treated for 9 weeks, once weekly, with estradiol dipropionate (5, 10, 15, 20, and 30 micrograms per animal) plus progesterone (in the range from 0.0 to 2.4 mg per animal). It was found that the higher the dosage of estradiol and progesterone used, the more complete the precopulatory behavioral pattern that was induced. Further, the same level of sexual responsiveness was achieved by various combinations of both hormones used. The dose-response relationships are less complicated in prepubertally ovariectomized females in comparison with those spayed as adults. Finally, the most complete pattern, characterized by ritualized darting, was not achieved at all.     

Impairment of spatial learning by estradiol treatment in female mice is attenuated by estradiol exposure during development

High doses of estradiol (E(2)) can impair spatial learning in the Morris water maze, in ovariectomized mice, but the same dose has no effect on adult castrated males. Here, we test the hypothesis that this sex difference is caused by neonatal actions of E(2). In Experiment 1, C57BL/6J pups were given daily estradiol benzoate (EB) or oil injections from the day of birth until postnatal Day 3. Adults were gonadectomized and received EB (s.c.) or oil 28 h before the first day of training, and 4 h before each of four daily training sessions on the Morris water maze. Females given oil as neonates, and EB prior to training displayed the poorest performance. Females that received EB as neonates and EB prior to training were insensitive to the deleterious effects of adult EB and performed better than males given the same hormone treatments. We conducted a second experiment using aromatase enzyme knockout (ArKO) mice. Adult male and female ArKO and wild-type (WT) littermates were gonadectomized and received either injections of oil or EB prior to and during water maze training (as described above). Hormone treatment failed to affect performance, yet, female but not male ArKO mice showed impaired learning compared to WT littermates. Thus, exposure to estradiol during neonatal development can counteract the deleterious effects of EB on adult spatial learning.     

Lipid peroxidation in ovariectomized and pinealectomized rats: the effects of estradiol and progesterone supplementation

In the present study, we investigated the effect of estradiol and progesterone supplementation on oxidant and antioxidant parameters of renal tissue in ovariectomized and pinealectomized rats. The study was carried out on 36 adult, Sprague-Dawley strain female rats, 6 months of age and weighing 200-250 g. The rats were divided into six groups, each group included six rats: Group 1: Sham-ovariectomized (Sham-Ovx); Group 2: Ovariectomized (Ovx); Group 3: Ovx and estradiol (E) and progesterone (P) supplemented (Ovx+E-P); Group 4: Ovariectomized and sham pinealectomy (Ovx+sham Pnx); Group 5: Ovariectomized+Pinealectomized (Ovx+Pnx); Group 6: Ovariectomized+Pinealectomized+Hormone Supplemented group (Ovx+Pnx+E-P). The levels of malondialdehyde (MDA), reduced glutathione (GSH) and glutathione peroxidase (GSH-Px) were analysed in renal tissues of rats. The highest and the lowest levels of MDA were determined in Groups 5 and 1 respectively (p < 0.001). However, GSH and GSH-Px levels demonstrated statistically important decreases in groups 2, 4, 5 (p < 0.001). The findings of this study demonstrate that ovariectomy leads to oxidative damage in renal tissue. Pinealectomy in addition to ovariectomy greatly increases the oxidative damage. However, female sex hormones supplementations to the Ovx and/or Ovx+Pnx rats protected against lipid peroxidation by activating the antioxidant system.     

Effects of testosterone and progesterone on the regression of Mullerian ducts induced by estradiol in the female chick embryo

An administration of testosterone or of progesterone to estradiol-treated female chick embryos increased the rate of those presenting a regression of their Müllerian ducts. This observation favoured the hypothesis according to which the regression induced by estrogens depends on the anti-Müllerian hormone of ovarian origin.