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Effects of progesterone and oestradiol on RNA and protein metabolism in the genital tract and on survival of embryos in the ovariectomized ewe.

The hormonal regulation of metabolism in the genital tract and the development of embryos during early pregnancy in the ewe have been examined. Ovariectomized ewes received injections of maintenance progesterone, oestrous oestradiol and priming progesterone according to schedules designed to simulate endogenous ovarian secretion during early pregnancy, around the time of oestrus and during the luteal phase of the oestrous cycle immediately preceding oestrus. The survival and development of embryos was dependent upon the dose of maintenaince progesterone and the duration of treatment at the time of transfer, but changes in progesterone dose did not change endometrial protein or RNA metabolism on particular days. Both priming progesterone and oestrous oestradiol were required for normal embryo development. Priming progesterone and oestrous oestradiol each increased endometrial RNA/DNA ratios during early pregnancy. There were no interactions between priming progesterone and oestrous oestradiol, their effects being simply additive. Neither maintenance nor priming progesterone had any effect on protein and RNA metabolism in the oviduct. It is suggested that in the intact ewe oestrogen secreted at oestrus and progesterone secreted prior to oestrus play important roles in the establishment of a uterine environment suitable for the subsequent normal development of embryos.

Animals↗

Effects of progesterone on the responses of Merino ewes to the introduction of rams during anoestrus.

The effects of progesterone on the responses of Merino ewes to the introduction of rams during anoestrus were investigated in two experiments. In the first experiment, the introduction of rams induced an increase in the levels of LH in entire ewes. The mean levels increased from 0.68 +/- 0.04 ng/ml (mean +/- s.e.m.) to 4.49 +/- 1.32 ng/ml within 20 min in ewes not treated with progesterone (n = 10). In ewes bearing progesterone implants that provided a peripheral concentration of about 1.5 ng progesterone per millilitre plasma, the LH response to the introduction of rams was not prevented, but was reduced in size so that the concentration was 1.38 +/- 0.15 ng/ml after 20 min (n = 5). Progesterone treatment begun either 2 days before or 6 h after the introduction of rams and maintained for 4 days prevented ovulation. In the second experiment ovariectomized ewes were used to investigate further the mechanism by which the ram evoked increases in tonic LH secretion. In ovariectomized ewes treated with oestradiol implants, the introduction of rams increased the frequency of the LH pulses and the basal level of LH. In the absence of oestradiol there was no significant change in pulse frequency but a small increase in basal levels. Progesterone again did not prevent but reduced the responses in ewes treated with oestradiol. It is suggested that following the withdrawal of progesterone treatment, the secretion of LH pulses in response to the ram effect would be dampened. This effect could be a component of the reported long delay between the introduction of rams and the preovulatory surge of LH in ewes treated with progesterone. Continued progesterone treatment prevented ovulation, probably by blocking positive feedback by oestradiol.

Animals↗

Suppression of arousal by progesterone in fetal sheep.

The high rate of progesterone synthesis by the placenta in late gestation exposes the ovine fetus to high concentrations of progesterone and its metabolites that may affect activity of the fetal brain. The aim of this study was to determine the effect of inhibiting maternal progesterone synthesis on sleep-wake activity in fetal sheep. Fetal and maternal vascular catheters, a fetal tracheal catheter, and electrodes for recording fetal electrocortical (ECoG), electro-ocular (EOG) and nuchal muscle electromyographic (EMG) activity were implanted. At 128-131 days gestation, progesterone production was inhibited by an injection of trilostane (50 mg), a 3beta-hydroxysteroid dehydrogenase inhibitor. Vehicle solution or progesterone (3 mg h(-1)) was then infused into the ewe between 6 and 12 h after the trilostane treatment. Maternal progesterone concentrations were significantly reduced from 1-24 h after trilostane treatment (P < 0.05) when followed by vehicle infusion. Fetal breathing movements (FBM), EOG, nuchal muscle EMG, and behavioural arousal increased 12 h after trilostane treatment (P < 0.05). In contrast, there was no change in fetal arousal, EOG, EMG or FBM activities when progesterone was infused after the trilostane treatment. These findings show that progesterone can influence fetal behaviour, and indicates that normal progesterone production tonically suppresses arousal, or wakefulness in the fetus.

3-Hydroxysteroid Dehydrogenases↗

Effects of luteectomy on the maintenance of pregnancy, circulating progesterone concentrations and lambing performance in sheep.

In sheep, there have been few and conflicting data regarding the necessity of the corpus luteum (CL) for the maintenance of pregnancy. The aims of the present study were to examine the effect of luteectomy on and after Day 50 of pregnancy on maternal plasma progesterone concentrations and the progression of pregnancy, to determine the minimum placental progesterone support required for the maintenance of pregnancy, and to evaluate the effect of luteectomy on lambing performance. In Experiment 1, four ewes luteectomized on Day 50 of pregnancy aborted 2-7 days after surgery, whereas pregnancy progressed and parturition occurred between Days 143 and 149, with live lambs, in three of four ewes and in four ewes luteectomized on Days 60 and 70 of pregnancy respectively. The mean (+/- SEM) progesterone concentrations on the day before and one day after luteectomy decreased from 4.87+/-0.85 to 0.42+/-0.06 ng mL(-1) (P<0.01), from 4.57+/-0.51 to 0.80+/-0.12 ng mL(-1) (P<0.02) and from 6.05+/-0.52 to 1.67+/-0.11 ng mL(-1) (P<0.01), respectively, for the ewes luteectomized on Days 50, 60 and 70 of pregnancy. The fall in progesterone concentrations was 90%, 80% and 71%, respectively, for the ewes luteectomized on Days 50, 60 and 70 of pregnancy. In Experiment 2, pregnancy progressed in four ewes luteectomized on Day 70 and parturition occurred between Days 146 and 149, with live lambs. The mean progesterone concentrations declined (P<0.01) from 6.9+/-0.7 ng mL(-1) on the day before luteectomy to 2.1 = 0.3 ng mL(-1) the day after surgery. The concentrations of progesterone in blood collected every 3 h during a 24-h period were stable on Days 60 and 80 of pregnancy, but they were lower (P<0.03) on Day 80 than on Day 60 of pregnancy, for each time period examined. In Experiment 3, the gestation length and birthweights of single, twin and triplet lambs were not different between the control intact ewes (n = 111) and the ewes luteectomized on Days 70-80 of pregnancy (n = 71). Lamb mortality was not different between the two groups (7.2% v. 8.4%, control v. luteectomized). In conclusion, these results showed that (1) the sheep CL is necessary to maintain pregnancy until at least Day 60, (2) progesterone withdrawal induced by luteectomy on and after Day 50 of pregnancy must be of a critical magnitude to provoke abortion, (3) after Day 60 of pregnancy, the CL and the placenta together secrete more progesterone than required for pregnancy maintenance, (4) there is no apparent 24-hour rhythm in maternal plasma progesterone concentrations before and after luteectomy, and (5) luteectomy at mid pregnancy has no apparent effect on gestation length, lamb birthweight or lamb mortality.

Animals↗

Peripheral plasma progesterone concentrations in pregnant and non-pregnant greyheaded flying-foxes (Pteropus poliocephalus) and little red flying-foxes (P. scapulatus).

Blood was collected from breeding-season and pregnant P. poliocephalus females shot in the wild and from captive pregnant and ovariectomized P. poliocephalus and P. scapulatus females. Peripheral plasma progesterone concentrations measured by radioimmunoassay were similar to those obtained by gas chromatography-mass spectroscopy: in intact non-pregnant P. poliocephalus females without corpora lutea (CLs) values ranged from 2 to 30 ng mL-1; after ovariectomy, they ranged from 1 to 85 ng mL-1. A significant source of progesterone in these bats may be the adrenal. In P. poliocephalus, peripheral plasma progesterone concentrations showed relatively little change over the breeding season or in early pregnancy when a CL formed, but increased from mid pregnancy to reach 200-800 ng mL-1 in late pregnancy. A mid-pregnancy ovary with CL contained 2.80 ng progesterone whereas the contralateral ovary contained 0.13 ng. Overall, CL size decreased during pregnancy and was negatively correlated with plasma progesterone concentrations. In late pregnancy, the main source of progesterone appears to be the placenta; plasma concentrations increase with placental growth and are significantly correlated with placental weight, and placentas contain 4-8 micrograms progesterone g-1. There was no evidence that progesterone concentrations fall before parturition. Limited observations indicated that peripheral progesterone concentrations follow similar patterns in P. scapulatus.

Animals↗

The classical progesterone receptor mediates Xenopus oocyte maturation through a nongenomic mechanism.

Xenopus laevis oocytes are physiologically arrested at G(2) of meiosis I. Resumption of meiosis, or oocyte maturation, is triggered by progesterone. Progesterone-induced Xenopus oocyte maturation is mediated via an extranuclear receptor and is independent of gene transcription. The identity of this extranuclear oocyte progesterone receptor (PR), however, has remained a longstanding problem. We have isolated the amphibian homologue of human PR from a Xenopus oocyte cDNA library. The cloned Xenopus progesterone receptor (xPR) functioned in heterologous cells as a progesterone-regulated transcription activator. However, endogenous xPR was excluded from the oocyte nucleus and instead appeared to be a cytosolic protein not associated with any membrane structures. Injection of xPR mRNA into Xenopus oocytes accelerated the progesterone-induced oocyte maturation and reduced the required concentrations of progesterone. In enucleated oocytes, xPR accelerated the progesterone-induced mitogen-activated protein kinase activation. These data suggest that xPR is the long sought after Xenopus oocyte receptor responsible for progesterone-induced oocyte maturation.

Animals↗

Monoclonal antibody to chicken oviduct progesterone receptor.

Antibodies to molybdate-stabilized chicken oviduct progesterone receptor were raised in a Wistar rat and detected by interaction with homogeneous radioiodinated progesterone receptor. Spleen cells of this rat were then fused with mouse Sp2/0-Ag14 myeloma cells and the antibodies produced by the hybrid cells were detected by double immunoprecipitation using the 125I-labeled receptor. Cells of one of the positive cultures were then cloned by limiting dilution and one hybridoma cell line was studied. The monoclonal antibody produced was an IgG2b, and it reacted with the molybdate-stabilized "8-9S" form of the chicken oviduct progesterone receptor, labeled with either [3H]progesterone or [3H]ORG 2058 (a high-affinity synthetic progestin). Kd for the 8-9S progesterone receptor was approximately equal to 1 nM. Progesterone receptor-monoclonal antibody complexes were labeled with radioactive progesterone, suggesting that antibody does not prevent hormone binding. By using a [35S]methionine-labeled antibody, we were able to detect the progesterone receptor independently of its characteristic function of binding radioactive hormone. No crossreaction with human progesterone receptor was detected.

Animals↗

Blocking of pregnancy in mice by immunization with anti-idiotype directed against monoclonal anti-progesterone antibody.

Passive transfer of monoclonal anti-progesterone antibodies shortly after mating blocks the onset of pregnancy in different species (mouse, rat, and ferret). Here we report that BALB/c mice can be actively immunized against progesterone, and hence against pregnancy, by means of rabbit anti-idiotypic antibodies specific for a mouse monoclonal anti-progesterone antibody, DB3. Some of the anti-idiotypic antibodies reacted with the steroid-combining site on the DB3 molecule. In response to repeated anti-idiotypic immunization, mice produced serum anti-progesterone antibodies (up to 100 micrograms/ml) that resembled DB3 in idiotypy, affinity, and specificity for progesterone and other steroid ligands. Thus an anti-idiotype can mimic the antigenicity of a steroid hormone with a high degree of accuracy. Compared with immunization with a progesterone-bovine serum albumin conjugate, the anti-progesterone response to anti-idiotype was considerably lower and clonally restricted. When mated after completion of the immunization course, the fertility rate of anti-idiotype-immunized mice was reduced to 30% from a control level of 91%. The anti-fertility effect was correlated with the circulating anti-progesterone concentration in individual animals and persisted for 4 or 5 estrous cycles. Active immunization with progesterone-bovine serum albumin was a highly effective means of rendering mice infertile; it reduced the fertility rate to zero over 16 or 17 estrous cycles. Our results suggest that anti-idiotypes may form the basis of contraceptive vaccines.

Animals↗

Serum progesterone levels in patients with gastric and colorectal cancers.

BACKGROUND: It is known that various malignant tissues possess progesterone receptors. It has been reported that the serum progesterone levels increase and show a prognostic significance in gastric carcinoma. We carried out a study to determine the serum progesterone level in gastric cancer, and colorectal cancer. METHODS: 140 of our patients were male and 56 female. We determined serum progesterone level in patients with gastric cancer, and colorectal cancer. Patients with benign diseases participated in the study as control group. RESULTS: In male patients with gastric cancer (n = 90) and colorectal cancer (n = 50) the serum progesterone level was not significantly higher than in the control group (n = 80) (mean: 1.08 +/- 0.73, 1.08 +/- 0.32 and 1.01 +/- 0.38 ng/ml, respectively). Among the female patients with gastric cancer (n = 26) and colorectal cancer (n = 30), the serum progesterone level was also not significantly higher than in the control group (n = 30) (mean: 1.19 +/- 0.77, 1.21 +/- 0.72 and 1.12 +/- 0.61 respectively). In males, the sensitivity of progesterone for gastric cancer was 55.5% and 64% for colorectal cancer with a specificity of 37.5%. In females, the sensitivity of progesterone for gastric cancer was 54% and 57% for colorectal patients with a specificity of 40%. CONCLUSIONS: Serum progesterone level does not correlate with the presence or absence of gastric cancer or colorectal cancer, and it is not a useful tumour marker.

Adenocarcinoma↗

Effect of micronized progesterone on bone turnover in postmenopausal women on estrogen replacement therapy.

PURPOSE: To determine whether daily oral micronized progesterone affects bone turnover, as estimated by serum and urine biochemical markers, in postmenopausal women on long-term estrogen replacement therapy (ERT). METHODS: We recruited 14 women aged 65 or older to participate in a 9-week trial with micronized progesterone. Each woman had undergone a hysterectomy and was on unopposed ERT at time of study entry. Women received micronized progesterone 100 mg twice daily in the first week and then received 200 mg twice daily in weeks 2-9. We measured markers of bone turnover in serum and urine collected at baseline and at 3 weeks, 6 weeks, and 9 weeks on treatment. Markers of bone formation were serum bone alkaline phosphatase (BAP), N-terminal procollagen peptides (PINP), and osteocalcin (OC). Markers of bone resorption were urinary cross-linked N-terminal and C-terminal telopeptides of type I collagen. In addition, we measured serum progesterone, estradiol and sex hormone binding globulin, triglycerides, total cholesterol, and high-density lipoprotein (HDL)-cholesterol levels at baseline and at 9 weeks on treatment. RESULTS: Mean serum progesterone levels increased from 1.6 +/- 1.1 to 15.2 +/- 3.9 ng/mL, which was within the luteal phase range (3-25 ng/mL). Crosslinked C-telopeptides of type I collagen and osteocalcin increased significantly (p < 0.05) with progesterone treatment, however, other bone markers did not change. Estradiol, estrone, and SHBG levels did not change with treatment. High-density lipoprotein-cholesterol levels decreased 19% (p < 0.001) at 9 weeks on treatment compared to baseline but total and low-density lipoprotein (LDL) cholesterol and triglycerides did not change with treatment. CONCLUSION: In postmenopausal women on long-term estrogen replacement therapy, micronized progesterone (400 mg/d) increased one marker each of bone resorption and bone formation. Other sensitive markers of bone turnover did not change with treatment. Further, micronized progesterone decreased HDL-cholesterol in these women. Our data do not support a beneficial effect of micronized progesterone on bone or cardiovascular risk factors in postmenopausal women.

Aged↗

A nongenomic mechanism for progesterone-mediated immunosuppression: inhibition of K+ channels, Ca2+ signaling, and gene expression in T lymphocytes.

The mechanism by which progesterone causes localized suppression of the immune response during pregnancy has remained elusive. Using human T lymphocytes and T cell lines, we show that progesterone, at concentrations found in the placenta, rapidly and reversibly blocks voltage-gated and calcium-activated K+ channels (KV and KCa, respectively), resulting in depolarization of the membrane potential. As a result, Ca2+ signaling and nuclear factor of activated T cells (NF-AT)-driven gene expression are inhibited. Progesterone acts distally to the initial steps of T cell receptor (TCR)-mediated signal transduction, since it blocks sustained Ca2+ signals after thapsigargin stimulation, as well as oscillatory Ca2+ signals, but not the Ca2+ transient after TCR stimulation. K+ channel blockade by progesterone is specific; other steroid hormones had little or no effect, although the progesterone antagonist RU 486 also blocked KV and KCa channels. Progesterone effectively blocked a broad spectrum of K+ channels, reducing both Kv1.3 and charybdotoxin-resistant components of KV current and KCa current in T cells, as well as blocking several cloned KV channels expressed in cell lines. Progesterone had little or no effect on a cloned voltage-gated Na+ channel, an inward rectifier K+ channel, or on lymphocyte Ca2+ and Cl- channels. We propose that direct inhibition of K+ channels in T cells by progesterone contributes to progesterone-induced immunosuppression.

Amino Acid Sequence↗

Serum progesterone levels correlate with decreased cerebral edema after traumatic brain injury in male rats.

Previous animal research suggests that progesterone may have powerful neuroprotective effects in traumatic brain injury (TBI). This experiment tested the hypothesis that progesterone levels correlate with decreased cerebral edema in male rats with bilateral medial frontal cortex injuries. Three groups of male Sprague-Dawley rats were used: injured given progesterone (4 mg/kg), injured given vehicle (oil), and uninjured controls given vehicle. Progesterone or vehicle was administered intraperitoneally at 1, 6, and 24 h postinjury. At 48 h postinjury, the rats were killed, brains extracted, and assayed for edema. Percent difference in water content of the area surrounding the lesion was compared to posterior cortex. A strong inverse relationship was found between serum progesterone levels and percent cerebral edema; the higher the progesterone levels, the lower the percent edema. Both progesterone and oil-treated animals had some edema compared to sham-operated controls. The brains of the injured animals given control solution were higher in water content than either the uninjured group or injured progesterone-treated rats 48 h postinjury. These findings confirm that progesterone significantly decreases cerebral edema after TBI in adult male subjects.

Animals↗

Nitric oxide reverses prostaglandin-induced inhibition in ovarian progesterone secretion in rats.

The present studies were undertaken to determine whether nitric oxide (NO) is involved in the regulation of ovarian progesterone and oestradiol secretion in rats. Immature female Sprague-Dawley rats at 27 days of age were injected s.c. with 4 IU pregnant mare's serum gonadotrophin (PMSG) and were killed 72 h after the injection. The ovaries were collected, weighed and cultured in Dulbecco's modified Eagle's medium containing saline, NO donor, NO synthesis inhibitor or prostaglandin F2 alpha (PGF2 alpha). After 24 h culture, the medium concentrations of progesterone and oestradiol were measured by radioimmunoassay. Results showed that: (i) diethylenetriamine (DETA)/NO (1 x 10(-6), 1 x 10(-5), 1 x 10(-4) M), an NO donor, caused a dose-dependent increase in progesterone synthesis (355 +/- 43, 443 +/- 46, 647 +/- 55 ng/g ovary respectively, P < 0.01) with a concomitant decrease in ovarian oestradiol secretion (408.1 +/- 50.7, 272.9 +/- 28.2, 132.6 +/- 34.6 pg/g ovary respectively, P < 0.01); (ii) neither progesterone nor oestradiol concentrations in the culture medium were altered by DETA without NO; (iii) NG-nitro-l-arginine methyl ester (1 x 10(-4) M), an inhibitor of NO synthesis, did not significantly affect progesterone and oestradiol secretion by rat ovaries; (iv) PGF2 alpha(1 x 10(-6) M) caused a fall in progesterone and oestradiol synthesis; (v) co-incubation with DETA/NO, significantly reversed the PGF2 alpha-induced decrease in progesterone concentrations from 184 +/- 29 to 388 +/- 60 ng/g (P < 0.01), but not that of oestradiol. It can be concluded that NO up-regulates progesterone secretion and down-regulates oestradiol secretion in rat ovaries, and NO can reverse PGF2 alpha-induced inhibition in ovarian progesterone secretion.

Animals↗

A prospective randomized study comparing intramuscular with intravaginal natural progesterone in programmed thaw cycles.

A simple programmed thaw cycle is described, during which the endometrium is prepared with oral oestradiol, followed by a natural progesterone source. This method involves minimal blood tests and uses inexpensive medications. Originally, an i.m. progesterone-in-oil preparation was used. Although highly successful in achieving high serum progesterone concentrations, the daily injections of the oily compound were found to be problematic from the patients' perspective. To examine the possibility of replacing the injectable progesterone with a vaginal preparation we conducted a prospective randomized study of 1 year's duration, during which time 170 and 184 thawing cycles were done with injectable and vaginal progesterone respectively. Although the progesterone blood concentrations obtained with the injectable preparation were more than twice those obtained with the vaginal progesterone, the clinical pregnancy rates (defined as percentage thawing cycles with gestational sac(s)/embryo transfer as demonstrated on ultrasound, 4 weeks after embryo transfer) were similar for both groups (15.9 and 16.8% respectively). Implantation and abortion rates were also comparable. These results agree with previous reports of higher uterine progesterone concentrations after the vaginal application. We conclude that the combination of oral oestradiol and vaginal progesterone is an effective, simple and inexpensive approach for programmed thaw cycles.

Administration, Intravaginal↗

Acrosome reaction induced in a limited population of human spermatozoa by progesterone (Ca(2+)-dependent) and ATP (Ca(2+)-independent).

We examined the kinetics of the acrosome reaction induced in human spermatozoa by progesterone (Ca(2+)-dependent) and adenosine 5'-triphosphate (ATP; Ca(2+)-independent). ATP and progesterone did not induce the acrosome reaction unless spermatozoa were incubated in a capacitation medium. ATP exhibited a constant induction of the acrosome reaction regardless of the incubation period, while progesterone began to induce the acrosome reaction after > or = 6 h of incubation. At 24 h of incubation, the percentages of spermatozoa in which the acrosome reaction had been induced by both progesterone and ATP were almost equal. To determine whether a limited population of human spermatozoa was reactive to both progesterone and ATP, we employed an MH61 bead binding method. When the acrosome-reacted spermatozoa were removed with MH61 beads, their percentage in the sperm suspension was decreased to < 3%. At 24 h of incubation, progesterone and ATP induced the acrosome reaction in 12.0 and 10.0% of spermatozoa, respectively. After MH61 bead binding was performed, the remaining spermatozoa did not react to the other activator (progesterone-->ATP, ATP-->progesterone). These findings indicate that only a limited population of human spermatozoa has the potential to undergo the acrosome reaction when stimulated by both progesterone and ATP.

Acrosome↗

Pre-ovulatory progesterone growth rate in patients treated with gonadotrophin-releasing hormone agonist and outcome of in-vitro fertilization.

The present study was undertaken to assess whether the increase in serum progesterone concentration following the administration of human chorionic gonadotrophin (HCG) may have predictive value on the in-vitro fertilization (IVF) success rate. Progesterone concentration on the day of HCG administration and the increase in progesterone concentration on the following day were evaluated in 140 consecutive patients undergoing IVF with embryo transfer. Stimulation protocol in all study patients entailed intranasal administration of short-acting gonadotrophin-releasing hormone agonist (GnRHa) buserelin and human menopausal gonadotrophin. A pregnancy rate of 37.2% was achieved when at least three embryos were transferred. The only significant difference between conception and non-conception cycles was found in serum progesterone concentrations after HCG administration (P < 0.01), whereas the mean progesterone concentration on the day of HCG did not differ. No difference in other hormonal or cycle parameters was observed. The increase in progesterone concentration was significantly greater in the group of patients who achieved pregnancy than in the group who did not (2.2 +/- 0.2 versus 1.6 +/- 0.1 ng/ml, respectively; P < 0.01). A critical breakpoint in serum progesterone was arbitrarily determined at 1 ng/ml. An increase in progesterone concentration > or = 1 ng/ml when three or more embryos were transferred was associated with a positive predictive value for pregnancy of 40.4% (sensitivity of 94.7%), whereas a negative predictive value of 86.7% was obtained when this value was < 1 ng/ml. These findings indicate that an adequate rise in serum progesterone following HCG administration provides useful information about the possible outcome of the treated cycle.

Adult↗

Effect of progesterone, mifepristone, and estrogen treatment during early pregnancy on conceptus development and uterine capacity in Swine.

A series of experiments was performed to investigate the influence of progesterone at Days 2 and 3 of pregnancy on conceptus development and uterine capacity. In experiment 1, unilaterally hysterectomized-ovariectomized (UHO) white crossbred gilts were given no treatment, estradiol valerate (5 mg given on Days 11 and 12), or progesterone (200 mg/day on Days 2 and 3 after mating). On Day 105 of pregnancy, each fetus and its associated placenta were weighed, and the number of live and dead fetuses was recorded for each litter. Early progesterone treatment reduced (P < 0.05) litter size (a measure of uterine capacity in UHO gilts). In experiment 2, intact white crossbred gilts were mated, given no treatment or progesterone treatment on Days 2 and 3 of pregnancy, and farrowed. Progesterone treatment decreased (P < 0.05) pregnancy rates. In pregnant gilts, progesterone had no effect on the number of live or stillborn piglets at birth, and gestation length was decreased (P < 0.05). Progesterone treatment did not affect the number of large or small piglets. In experiment 3, intact gilts were mated at estrus and then received 1). no treatment or treatment with 2). 100 mg, 3). 200 mg, or 4). 400 mg mifepristone (also known as RU486) on Day 2 of pregnancy. On Day 11 of pregnancy, both uterine horns were flushed, the number and diameter of each conceptus was recorded, and the flushed material was assayed for total protein and acid phosphatase. The 400 mg mifepristone treatment decreased conceptus diameter (P < 0.05) and total protein (P = 0.06) in the uterine flushings. In experiment 4, UHO gilts were mated at estrus, injected with either corn oil (control) or mifepristone (400 mg) on Day 2 of pregnancy, and killed on Day 105 of pregnancy, and the number and weight of live fetuses and placentas was recorded. In contrast to the effect of progesterone treatment, mifepristone decreased uterine capacity by decreasing the number of small conceptuses. These data suggest that progesterone concentrations on Days 2 and 3 of pregnancy in swine influence the rate of conceptus development during early pregnancy and uterine capacity during later pregnancy.

Animals↗

Sufficient progesterone-priming prior to estradiol stimulation is required for optimal induction of the cervical prostaglandin system in pregnant sheep at 0.7 gestations.

The purposes of this study were to determine the separate and interactive functions of progesterone and estradiol in regulating the cervical prostaglandin (PG) system in pregnant sheep at 0.7 gestations. At 106-108 days of gestational age (dGA), ewes were treated with vehicle for 14 days (n = 5) or vehicle for 12 days followed by estradiol 5 mg twice a day, intramuscularly for 2 days (n = 5) or progesterone 100 mg, twice a day, intramuscularly for 14 days (n = 5) or progesterone 100 mg twice a day, intramuscularly for 10 days and then 2 days vehicle followed by estradiol 5 mg twice a day intramuscularly for 2 days (n = 5). At 121-123 dGA, cervical tissues were obtained under halothane anesthesia. Cervical RNA and protein were extracted and analyzed for prostaglandin-endoperoxide synthase 2 (COX2), two PGE(2) receptors, PTGER2 and PTGER4, and estrogen receptor alpha (ESR1) by Northern and Western blot analysis. Immunocytochemistry and in situ hybridization were applied to localize cellular distribution of COX2, PTGER2, and PTGER4 in the cervix. Data were analyzed by ANOVA. COX2 and PTGER4 mRNAs and proteins were increased (P < 0.05) in ewes treated with combined estradiol and progesterone but not in ewes treated with estradiol or progesterone alone compared with controls. ESR1 mRNA was increased in ewes treated with progesterone and estradiol plus progesterone. In contrast, PTGER2 mRNA and protein remained the same after all treatments. COX2 mRNA and protein were localized only in cervical glandular epithelial cells, whereas PTGER2 and PTGER4 were localized in both cervical glandular epithelial and smooth muscle cells. In conclusion, these data suggest that additional progesterone priming at 0.7 gestations synergizes with estradiol to induce cervical COX2, PTGER4, and ESR1 and support our hypothesis that stimulation of the cervical PG system by estradiol is optimized by sufficient progesterone priming in the pregnant sheep cervix.

Animals↗