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The control of progesterone secretion during the estrous cycle and early pseudopregnancy in the rat: prolactin, gonadotropin and steroid levels associated with rescue of the corpus luteum of pseudopregnancy.

The hormonal factors associated with converting a corpus luteum of estrous cycle into a corpus luteum of pseudopregnancy were studied by measuring LH and FSH prolactin, estradiol and progesterone levels in decapitated rats during the 4-day estrous cycle and a comparable time of pseudopregnancy (lights on 0600-0800 hr.). During the estrous cycle, prolactin, LH and FSH remained low and unchanging except on the afternoon of proestrus, when typical proestrous surges were observed. In contrast, estradiol levels began to increase on D-1, from baseline values of 7 pg/ml to approximately 15-20 pg/ml. These levels were maintained until the afternoon of D-2 when estradiol further increased to reach peak levels of 40-50 pg/ml by 0900 hr on proestrus. Estradiol then declined in relation to the increase in LH secreation and had returned to baseline by estrus. Progesterone secretion by the corpora lutea of the cycle also increased on the afternoon of D-1 and reached a maximum value of 25-30 ng/ml early on the morning of D-2. At this time, a precipitious fall in progesterone occurred, returning to baseline values of 5-1- ng/ml by 0700 on D-2 signifying the regression of the corpora lutea of the cycle. Progesterone remained low thereafter until the afternoon of proestrus when levels increased in response to the proestrus when levels increased in response to the proestrous surge of LH. Following cervical stimulation at 1900 hr on proestrus, no differences were noted, with respect to the estrous cycle, in LH, FSH or estradiol secreation through the afternoon of D-2. Surprisingly, progesterone levels did not differ in the cycle and pseudopregnancy until the early morning of D-29 instead of progesterone levels falling to baseline as they had during the cycle, the corpora lutea of pseudopregnancy were rescused, progesterone increasing dramatically to reach levels of 45-50 ng/ml by 1700 hr on that same day. The only difference in hormone secretion that was noted which could account for this marked divergence in progesterone secretion was the pattern of prolactin secretion following cervical stimulation. In contrast to the low levels seen during the estrous cycle, biphasio surges of prolactin secretion occured each day, one being nocturnal (0100-0900 hr) and the other diurnal (1500-2100 hr). The rescue of the corpus luteum occured in association with the nocturnal surge on D-2. These results suggest that nocturnal surge on D-2, PROLACTIN IS THE MAJOR Luteotropic stimulus which transforms and estrous cycle into pseudopregnancy by prolonging progesterone secretion from the corpus luteum. Moreover, if LH is important for progesterone secretion, no changes were observed in the pattern of LH secretion which can account for the rescue of the corpus luteum.

Animals↗

Plasma concentrations of progesterone and 13,14-dihydro-15-keto prostaglandin F-2 alpha in pregnant, pseudopregnant and hysterectomized pseudopregnant mice.

Hysterectomized pseudopregnant mice had plasma progesterone concentrations on Days 6, 8, 10 or 12 after mating that were greater (P less than 0.05) than those of intact pseudopregnant mice and lower (P less than 0.01) than those of pregnant mice. Concentrations of 13,14-dihydro-15-keto-prostaglandin F-2 alpha on Days 6, 7, 8, 9 or 10 after mating were lower (P less than 0.05) in pregnant and higher (P less than 0.01) in intact pseudopregnant mice than in hysterectomized pseudopregnant mice. The results support the conclusion that there are uterine and extra-uterine luteolytic mechanisms which are mediated by prostaglandin and are perhaps functional by Day 6 of pregnancy.

Animals↗

Prolactin and delayed pseudopregnancy in the rat.

In (RXU)F1 hybrid rats delayed pseudopregnancy was induced in three different ways: 1) by removal of all recent corporl lutea on day 2 of pseudopregnancy, 2) by removal of the in situ ovaries from ovarian graft-bearing animals on day 0 of pseudopregnancy, 3) by administration of 1 mg of ergocornine hydrogenmaleinate (ECO) on day 1 of pseudopregnancy. These procedures ended pseudopregnancy and in 50-60% of the animals a delayed pseudopregnancy with a duration of 6-15 days was observed after the experimental cycle. After sterile copulation two daily prolactin peaks were observed. Removal of the ovaries in situ (from ovarian graft-bearing animals) or of recently formed corpora lutea (from non-grafted animals) caused the disappearance of the prolactin peak at 19.00 h, without affecting the occurrence of the 03.00 h peak. The administration of ECO caused the disappearance of both prolactin peaks. Until the day of estrus prior to delayed pseudopregnancy there were no differences in prolactin concentrations at 03.00 or 19.00 h between animals which became delayed pseudopregnant and those which remained cyclic. In the latter animals the 03.00 h surges decreased slowly in the luteectomized and the ovariectomized animals and were no longer present 7-9 days after copulation. In animals becoming delayed pseudopregnant both prolactin peaks were present from day 0 of delayed pseudopregnancy onwards. Progesterone cencentrations during delayed pseudopregnancy were relatively low when delayed pseudopregnancy had a duration of 6-9 days. When delayed pseudopregnancy lasted 10 days or more normal progesterone values were found.

Animals↗

Effect of an oxytocin receptor antagonist on ovarian and uterine synthesis and release of prostaglandin F2 alpha in pseudopregnant rats.

There is substantial experimental evidence suggesting that oxytocin has a role in luteolysis in ruminates. Endogenous pulses of uterine prostaglandin (PG) F2 alpha occur synchronously with pulses of oxytocin during luteolysis; leading us to propose a possible feedback loop between uterine PGF2 alpha and luteal oxytocin. In rates, the mechanism whereby oxytocin acts has not been well elucidated. In the present report, the effects of an oxytocin receptor antagonist in pseudopregnant rats were investigated. Pseudopregnancy was induced in immature female rats by gonadotrophin treatment; this resulted in the formation of corpus luteum that remained functional for 9 +/- 1 days. The pseudopregnant rats were assigned to one of the following four groups. In the first group the relationship between the release of ovarian and uterine PGF2 alpha was tested. We also studied the serum progesterone during the pseudopregnancy. We found that PGF2 alpha released into the incubation medium from ovaries of pseudopregnant rats increased (p < 0.05) and was maximal on day 9 of pseudopregnancy. This concentration remained high until day 10 of pseudopregnancy and then decreased. The PGF2 alpha released from the uterus to the incubation medium rose (p < 0.05) on day 8 of pseudopregnancy and reached the peak value on day 10. the serum progesterone was increased (p < 0.001) on day 2 pseudopregnancy and was greater on day 5 (p < 0.001). The second and third group received a specific oxytocin receptor antagonist (1-deamino-2-O-methyltyrosine) in two different concentrations (0.05 or 0.2 mumol/l before the peak of PG release. Both doses employed decreased (p < 0.001) the release into the incubating medium of PGF2 alpha from ovaries and uterus. Indeed, after the treatment, the progesterone levels were higher (p < 0.001) than control on day 10 of pseudopregnancy. In the fourth group, a potent inhibitor of cyclooxygenase activity was administered on day 8 of pseudopregnancy into the ovarian bursa. The serum progesterone levels increased (p < 0.01) compared to control suggesting a possible role of ovarian PG in the luteolytic phase of the corpus luteum regression. Thus, our findings show that oxytocin is luteolytic in pseudopregnant rats and this action is mediated by oxytocin receptors, as it was blocked by a specific oxytocin receptor antagonist.

Animals↗

Plasma concentrations of prolactin in overtly pseudopregnant Afghan hounds and the effect of metergoline.

The effect of metergoline, a 5-hydroxytryptamine (serotonin) antagonist, on the plasma concentrations of prolactin in overtly pseudopregnant Afghan hounds and on the clinical symptoms of overt pseudopregnancy were studied. Plasma concentrations of prolactin and progesterone were determined in six Afghan hounds with signs of overt pseudopregnancy for 2-3 weeks and in three Afghan hounds that were not pseudopregnant at the time of blood sampling. In the overtly pseudopregnant bitches the plasma concentrations of prolactin before treatment (35.5 +/- 8.5 micrograms l-1) were significantly higher than the plasma concentrations of prolactin of the three bitches that were not pseudopregnant (6.3 +/- 0.5 micrograms l-1); the latter values were similar to those of non-pseudopregnant beagle bitches during the total luteal phase. The six pseudopregnant Afghan hounds were treated for 10 days with the antiserotoninergic drug metergoline. At 2 h after the onset of treatment with metergoline, the mean plasma concentration of prolactin had decreased to 10.8 +/- 2.9 micrograms l-1. The plasma concentrations of prolactin continued to decline to 5.4 +/- 1.0 micrograms l-1 at 4 h and to 1.0 +/- 0.1 microgram l-1 during treatment days 3-10. Signs of pseudopregnancy, such as swelling of the mammary glands and digging, decreased during the treatment period. The treatment was associated with mild behavioural side effects such as whimpering and aggressiveness. These side effects are probably not related to suppression of prolactin but are due to a direct effect on serotoninergic pathways in the brain. It is concluded that high plasma concentrations of prolactin are associated with the development and maintenance of pseudopregnancy. The serotonin antagonist metergoline strongly suppresses plasma concentrations of prolactin in pseudopregnant dogs and decreases the clinical signs of pseudopregnancy.

Animals↗

Evidence for multiple uterine modulators of rabbit luteal function: the effect of hysterectomy during pseudopregnancy in the estrogen-treated rabbit.

Previous studies show that hysterectomy on Day 1 of pseudopregnancy prolongs serum progesterone secretion in estrogen-treated pseudopregnant rabbits. These studies were undertaken to determine the day of pseudopregnancy when uterine factors are released to alter luteal function. When hysterectomies were performed on either Day 5, 8, 10, or 13 of pseudopregnancy, serum progesterone concentrations were greater than 10 ng/ml between Days 18 and 27 of pseudopregnancy compared to levels of approximately 4 ng/ml in sham-hysterectomized rabbits on these same days. In contrast, serum progesterone levels were not elevated when hysterectomies were performed on Day 11 of pseudopregnancy and were only partially maintained when hysterectomies were performed on Day 12 of pseudopregnancy. Twice daily injections of prolactin (1.5 mg, s.c.) between Days 1 and 33 of pseudopregnancy were unable to mimic the effect of estradiol in the hysterectomized rabbit. Twice daily injections of indomethacin (8 mg/kg, s.c.) between Days 6 and 23 of pseudopregnancy lowered uterine and luteal prostaglandin F2 alpha levels approximately 10-fold on Day 24 of pseudopregnancy but did not maintain progesterone secretion. Serum cholesterol levels were not altered by hysterectomy on any day and were thus not related to the maintenance of progesterone production. These results suggest that the uterus produces both inhibitory and stimulatory factors that effect luteal progesterone secretion. First, an inhibitor is released between Days 10 and 11 of pseudopregnancy in estrogen-treated rabbits that prevents the rabbit corpus luteum from responding to estradiol.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Luteolytic and antiluteolytic effect of the antiprogestagen RU486 in pseudopregnant rats.

To study the effects of the antiprogestagen RU486 on luteal activity in pseudopregnant rats, adult female rats made pseudopregnant by sterile copulation were given daily injections with oil vehicle or with RU486 (2 mg/day) either during the entire period of pseudopregnancy (day 1 till day 14) or during the second half of pseudopregnancy (day 8 till day 14). Blood was taken every other day to measure serum concentrations of progesterone. At autopsy, on day 15, the weights of ovaries, isolated corpora lutea and pituitary glands were recorded. In a second study using the same experimental protocol, blood was taken via a jugular vein cannula on days 8, 9, 10 and 11 after induction of pseudopregnancy; on each of these days blood samples were taken at 0700, 0800 and 0900 h, and at 1700, 1800 and 1900 h to measure plasma concentrations of prolactin, LH and progesterone. Administration of RU486 from day 1 of pseudopregnancy onwards had no effect on the increasing concentrations of serum progesterone during the first half of pseudopregnancy. Thereafter progesterone concentrations of serum progesterone during the first half of pseudopregnancy. Thereafter progesterone concentrations increased further in RU486-treated rats whereas they decreased in oil-treated pseudopregnant rats. Administration of RU486 from day 8 of pseudopregnancy onwards resulted in a decline in progesterone concentrations in serum on day 10 followed by ovulation on day 11. Plasma LH concentrations in rats treated with RU486 from day 1 of pseudopregnancy were higher than those in oil-treated rats on days 8, 9, 10 and 11.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

[Plasma prolactin concentration and the effect of metergoline in pseudopregnant Afghan hounds].

The effects of metergoline, a 5-hydroxytryptamine (serotinin) antagonist, on the plasma concentrations of prolactin in overtly pseudopregnant Afghan hounds and on the clinical symptoms of overt pseudopregnancy were studied. Plasma concentrations of prolactin and progesterone were determined in six Afghan hounds with signs of overt pseudopregnancy for 2-3 weeks and in three Afghan hounds that were not pseudopregnant at the time of blood sampling. In the overtly pseudopregnant bitches the plasma concentrations of prolactin before treatment (35.5 +/- 8.5 micrograms l-1) were significantly higher than the plasma concentrations of prolactin of the three bitches that were not pseudopregnant (6.3 +/- 0.5 micrograms l-1); the latter values were similar to those of non-psueodopregnant beagle bitches during the total luteal phase. The six pseudopregnant Afghan hounds were treated for 10 days with the antiserotoninergic drug metergoline. At 2 h after the onset of treatment with metergoline, the mean plasma concentration of prolactin had decreased to 10.8 +/- 2.9 micrograms l-1. The plasma concentrations of prolactin continued to decline to 5.4 +/- 1.0 micrograms l-1 at 4 h and to 1.0 +/- 0.1 microgram l-1 during treatment days 3-10. Signs of pseudopregnancy, such as swelling of the mammary glands and digging, decreased during the treatment period. The treatment was associated with mild behavioural side effects such as whimpering and aggressiveness. These side effects are probably not related to suppression of prolactin but are due to a direct effect on serotoninergic pathways in the brain. It is concluded that high plasma concentrations of prolactin are associated with the development and maintenance of pseudopregnancy. The serotonin antagonist metergoline strongly suppresses plasma concentration of prolactine in pseudopregnant dogs and decreases the clinical signs of pseudopregnancy.

Animals↗

Dioestrous ovariectomy: a model to study the role of progesterone in the onset of canine pseudopregnancy.

It has been suggested that overt pseudopregnancy in bitches is caused by an increase in the concentration of serum prolactin as a result of an abrupt decrease in progesterone concentration in the late luteal phase. This hypothesis was tested by using ovariectomy at dioestrus as an experimental model. A total of 18 intact cross- and purebred bitches were used. Eleven animals were ovariectomized (day 0) between day 25 and day 40 of the oestrous cycle, and seven intact bitches were used as controls. Blood samples for determination of prolactin and progesterone concentrations were collected on days -1, 1, 2, 3 and 7 in the ovariectomized group, and on day 1 and day 7 in the control group. On day 7, the presence or absence of overt pseudopregnancy was recorded. The four ovariectomized bitches with a history of pseudopregnancy showed signs of overt pseudopregnancy (P < 0.01). On day 7, progesterone concentrations were significantly higher in the control than in the ovariectomized bitches (P < 0.01). The expected decrease in serum progesterone concentration after ovariectomy was similar in pseudopregnant bitches and non-pseudopregnant bitches. However, in pseudopregnant bitches, but not in non-pseudopregnant bitches, there was a marked increase (expressed as percentage change) in the concentration of prolactin between day -1 and day 7 (P < 0.01). It was concluded that the abrupt decrease in progesterone concentrations does not lead systematically to pseudopregnancy. Only in bitches predisposed to pseudopregnancy would an abrupt decrease in progesterone concentrations induce a substantial increase in prolactin concentrations, which in turn would trigger the typical signs of pseudopregnancy.

Animals↗

The effect of pseudopregnancy on glomerular filtration rate and salt and water reabsorption in the rat.

1. Glomerular filtration rate (G.F.R.) and salt and water reabsorption were measured in age-matched virgin rats and rats at different stages of pseudopregnancy and post-pseudopregnancy. 2. Tubular reabsorption and G.F.R. were significantly higher in later pseudopregnancy. Values at mid-pseudopregnancy were intermediate between virgin controls and late-pseudopregnancy. In post-pseudopregnancy G.F.R. and reabsorption had returned to values not different from the virgins. 3. Expansion of extracellular fluid volume (e.c.f.v.) and elongation of proximal tubules were observed during pseudopregnancy. In post-pseudopregnancy increased tubular length was still apparent but e.c.f.v. was not. 4. The remarkable similarity in the changes in e.c.f.v. and renal functions and structure during pseudopregnancy to those in early pregnancy suggests that the feto-placental unit is not necessary for the pregnancy changes. 5. The differences between the time course of change in plasma progesterone and the time courses of changes in e.c.f.v., renal functions and tubular morphology in late pseudopregnancy suggest that progesterone is not directly involved.

Absorption↗

Changes in lipid peroxide and antioxidant enzyme activities in corpora lutea during pseudopregnancy in rats.

This study investigated the involvement of lipid peroxidation and antioxidant enzymes in the regulation of luteal function in pseudopregnant rats. The activities of superoxide dismutase (SOD), a specific scavenger of superoxide radicals, and glutathione peroxidase, a scavenger of hydrogen peroxide, and lipid peroxide concentrations were measured in the corpus luteum on days 1, 3, 5, 7, 9, 11 and 13 of pseudopregnancy. The activity of SOD in the corpus luteum gradually increased until day 9 of pseudopregnancy and decreased thereafter, in a similar manner to serum progesterone concentration. Glutathione peroxidase activity significantly increased from day 1 to day 3 and remained high until day 11 of pseudopregnancy. The concentrations of lipid peroxides in the corpus luteum increased from day 3 to day 13 of pseudopregnancy. The involvement of prostaglandin F2 alpha (PGF2 alpha) in the production of lipid peroxides in regression of the corpus luteum was investigated by administering PGF2 alpha (3 mg kg-1, s.c.) or saline on days 7, 9 and 12 of pseudopregnancy. Each group of rats was autopsied 2 h later, and SOD activity, glutathione peroxidase activity and the concentration of lipid peroxides in the corpus luteum were determined. PGF2 alpha significantly increased lipid peroxide concentrations in the corpus luteum on days 7, 9 and 12 of pseudopregnancy (approximately twofold increases on days 7 and 9, and a fivefold increase on day 12, compared with the control that received saline). The activity of SOD in the corpus luteum was significantly increased by PGF2 alpha on days 7 and 9, but not on day 12, of pseudopregnancy. PGF2 alpha did not cause any significant changes in glutathione peroxidase activity in the corpus luteum on days 7, 9 and 12 of pseudopregnancy. It is concluded that lipid peroxides play an important role in regulating luteal function in pseudopregnant rats.

Animals↗

Effects of droloxifene on apoptosis and Bax, Bcl-2 protein expression of luteal cells in pseudopregnant rats.

AIM: To study the effects of droloxifene on apoptosis and the expression of Bax and Bcl-2 protein in corpus luteum of pseudopregnant rats. METHODS: HE staining was used to examine the histological changes of ovaries. Apoptosis detection in situ was performed with TUNEL method. Expression of Bax and Bcl-2 protein was observed by immunohistochemistry analysis. RESULTS: Apoptosis of luteal cells during the spontaneous regression of corpus luteum of pseudopregnant rats appeared on d 13 of pseudopregnancy, and the marked increase of apoptotic luteal cells could be observed on d 15. When pseudopregnant rats were treated with droloxifene 20 mg . Kg on d 2, apoptosis of luteal cells could be observed on d 8 and the duration of pseudopregnancy could be shortened from (15.5 +\- 1.1) d to (12.8 +\- 1.6) d. In pseudopregnant rats, the expression of Bax and Bcl-2 protein was found in the cytoplasm of luteal cells. However, no obvious differences in the intensity or localization could be found during various days of the pseudopregnancy, while an increase in Bax and a decrease in Bcl-2 protein expression could be induced by droloxifene treatment. CONCLUSION: Droloxifene could facilitate apoptosis of luteal cells in pseudopregnant rats and shorten the period of pseudopregnancy. An increased Bax/Bcl-2 ratio might be involved in the facilitation of apoptosis induced by droloxifene in corpus luteum of pseudopregnant rats.

Animals↗

Estrogen-induced pseudopregnancy in gilts: Its use in estrus synchronization and subsequent influence on litter response.

The effectiveness of using estrogen-induced pseudopregnancy followed by PGF(2alpha) (Lutalyse) treatment to synchronize estrus in gilts and the subsequent effect of pseudopregnancy on litter parameters was examined in two experiments. Experiment 1 consisted of a control (n = 49) group and a pseudopregnant (n = 49) group. Pseudopregnant gilts received PGF(2alpha) between Days 25 and 38 and were bred at the ensuing estrus. Ninety-two percent of the gilts responded to PGF(2alpha) and exhibited estrus in 4.9 +/- 0.2 (Least Square Mean +/- SEM) days. Duration of estrus was longer in pseudopregnant compared to control gilts (2.8 vs 1.7 d, P<0.001). Experiment 2 consisted of a control (n = 24) group and a pseudopregnant (n = 24) group. All pseudopregnant gilts received PGF(2alpha) on Day 38 and 79% responded with a mean onset of estrus at 5.2 +/- 0.2 d. As in Experiment 1, duration of estrus was longer in gilts from the pseudopregnant group (2.2 vs 1.7 d, P<0.005). The percentage of farrowing was similar for both groups in both experiments, and no differences existed in the number of pigs born and those alive at 21 d post partum. Litters from gilts in the pseudopregnant groups were heavier at birth than those in the control group (Experiment 1, 17.21 vs 15.25 kg, P<0.01; Experiment 2, 15.31 vs 13.55 kg, P<0.02) and at 21 d (Experiment 1, 60.81 vs 56.79 kg, P<0.01; Experiment 2, 51.89 vs 46.96 kg, P<0.02). Survival rate at 21 d was higher in offspring from pseudopregnant gilts in both experiments (Experiment 1, 85.3% vs 78.2%, Experiment 2, 91.4% vs 81.2%).

Journal Article↗

Elevated progesterone during pseudopregnancy may prevent bone loss associated with low estrogen.

There is increasing evidence for a role of endogenous progesterone in mineral and skeletal metabolism. The purpose of this study was to compare skeletal changes that occur during a condition of high endogenous progesterone but low estrogen (pseudopregnancy) with a condition of low endogenous progesterone and low estrogen (ovariectomy). Pseudopregnancy was selected over pregnancy to eliminate placental factors that may influence mineral metabolism. Rats were ovariectomized (OVX) or pseudopregnancy initiated, and bones were collected 13 days later. In some animals, blood was collected by indwelling catheters for determination of progesterone and estrogen levels. At mid-pseudopregnancy, there were substantial elevations in progesterone but estrogen was below the level of detection. Progesterone and estrogen were below the level of detection in the OVX rats. Longitudinal growth rates were increased compared with the normal cycling rats in both the pseudopregnant and OVX groups, indicative of decreased estrogen levels, but they were greatest in the OVX rats. Cancellous bone mass was maintained in the pseudopregnant rats compared with normal cycling rats but significantly reduced in the OVX animals. As expected, increased bone formation and turnover rates were observed in the OVX animals although some indices of bone formation were also increased in the pseudopregnant animals. Osteoclasts were significantly increased in the OVX but not the pseudopregnant animals compared with normal cycling rats. Increased periosteal bone formation indices are known to occur following OVX, but the greatest periosteal formation rates were observed in the pseudopregnant animals. While possible roles for some other endocrine agents cannot be excluded at this time, the data from the present study suggest that endogenous progesterone may have a role in the maintenance of bone mass perhaps by decreasing bone resorption while maintaining or increasing bone formation during physiological periods of low estrogen such as occurs during early pregnancy.

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