Studies on the role of zinc in parturition in the rat.

1. Pregnant rats were fed either low (less than 1 p.p.m.) Zn or control (40 p.p.m. Zn) diets from day 10 of gestation. They were killed at intervals during the last 96 h preceding the normal time for onset of parturition, and differences in plasma progesterone, oestradiol-17 beta and ovarian 20 alpha-hydroxysteroid dehydrogenase were assessed. 2. Gestation was prolonged in Zn-deficient rats. 3. Although the preparturient decline in plasma progesterone began at the same time in all groups, at term, plasma progesterone concentration in Zn-deficient rats remained significantly higher than in normal females. 4. Induction of ovarian 20 alpha-hydroxysteroid dehydrogenase activity was delayed by about 8 h by Zn deficiency. This delay was not observed if prostaglandin F2 alpha was injected previously. 5. The results suggest a Zn-dependent step(s) in uterine synthesis and/or release of prostanoids.     

Progesterone, 20α-dihydroprogesterone and PGF; Interrelated hormonal changes during pseudo-pregnancy in the rat

Ovarian and uterine tissue concentrations of progesterone, 20α-dihydroprogesterone (20_αOHP) and prostaglandin F (PGF) were measured during hormonally-induced pseudopregnancy, as were plasma levels of progesterone and 20αOHP, in hysterectomised and sham-operated rats. Elevated levels of PGF in uterine and ovarian tissues were coincident with declining concentrations of progesterone and increasing concentrations of 20αOHP in the sham-operated rats. Maximum PGF concentrations were apparent in uterine tissue 14 days after hCG injection, coincident with the plasma, ovarian and uterine nadir concentrations of progesterone. A small but statistically significant increase in ovarian PGF was apparent at this time in sham-operated rats. This elevation of ovarian PGF was abolished by hysterectomy.     

Plasma gonadotropins and prolactin in pseudopregnancy in the rat

Radioimmunoassay presented a method of measuring plasma levels of FSH,LH and prolactin in pseudopregnant rats. Plasma prolactin levels doubled 15 minutes following cervical probing (p .01) on the day of estrus. Plasma LH levels were not significantly elevated. Due to the use of ether anesthesia at blood collecting 3 hr before and 15 minutes after stimulation, only 1 of 16 rats developed pseudopregnancy. On Day 4 of pseudopregnancy in rats mated with vasectomized males; plasma LH was lower (p .05) than in normal rats on the first day of diestrus, perhaps due to the suppressive action of ovarian progesterone and some estrogen. FSH was higher than in normal rats (p .05) perhaps due to the lesser sensitivity of FSH to the inhibitory effect of progesterone. Large decidoumata developed by Day 9 in uterine horns traumatized on Day 4 (153 plus or minus 8 mg uterus weight compared to 1510 plus or minus 204 mg). Thus, the corpora remain functional after LH and prolactin are at normal and subnormal levels. On Day 9 plasma prolactin was lower than at Day 1 of diestrus (p .001). Plasma FSH was elevated (p .01). Plasma LH was unchanged. The degree of rise of LH levels 5 days following ovariectomy on Day 4 of psuedopregnancy or on the first day of diestrus was greater in the former group (p .02), perhaps due to rebound of LH from suppression by ovarian steroids. FSH rose equally in both groups. Prolactin remain about the same. Increased prolactin release by the adenohypophysis was briefer than expected.     

Gonadotropin-releasing hormone (GnRH) inhibits ovulation induced with luteinizing hormone (LH) in proestrous hypophysectomized rats

In this paper we present evidence that a single low dose of the natural synthetic gonadotropin-releasing hormone (GnRH), inhibits ovulation induced by LH in proestrous-hypophysectomized rats. Rats hypophysectomized by the parapharyngeal route in the morning of proestrus received an intravenous injection of 100 or 300 ng GnRH at 1400 h immediately followed by 1.0 microgram LH per 100 g bw. In control groups, either one or both hormones were replaced with 0.9% NaCl. Ovulation was assessed the following morning by counting the ova present in oviductal flushings. All the rats treated with LH alone ovulated, and the addition of GnRH reduced significantly the number of ovulating rats and the number of ova per ovulating rat. In other groups of rats hypophysectomized in the morning of proestrus and treated in the same way, ovarian or adrenal secretory rates of estradiol and/or progesterone were measured after cannulation of the corresponding vein, in the afternoon of proestrus. In these animals, GnRH failed to inhibit either the ovarian progesterone surge observed 2 h after LH administration, or the adrenal progesterone secretion. All hypophysectomized rats showed lower ovarian secretory rate of estradiol than intact rats; this rate was not affected by treatment with LH or LH plus GnRH. The systemic estradiol levels in plasma of hypophysectomized rats were distributed within a range of 20 pg/ml to 50 pg/ml. The number of rats whose levels were above 21 pg/ml on estrus day was significantly higher in rats receiving 300 ng GnRH as compared to those receiving 100 ng GnRH, reaching values that surpassed the concentration found in intact, untreated animals at the same time of estrus. This effect did not depend on LH administration.     

Anti-fertility effects of embelin in female Sprague-Dawley rats may be due to suppression of ovarian function

Effects of embelin on oestrous cycle, plasma levels of progesterone and oestradiol, and in vitro production of oestradiol and progesterone by mixed ovarian cells was studied. Forty adult (4 months old) regularly cycling female Sprague-Dawley rats were divided into four groups of 10 rats each. Groups I and II (controls) were given 1 ml/kg body weight of physiological saline or corn oil (vehicle). Groups III and IV received 10 mg/kg and 20 mg/kg body weight embelin in corn oil, respectively. Emberlin disrupted the oestrous cycles in Groups III and IV animals, and there was a significant depression in plasma oestradiol (p <0.05) and progesterone (p <0.02) at both 10 and 20 mg/kg body weights, respectively. Isolated mixed ovarian cells from embelin treated rats produced significantly less progesterone and estradiol than controls in vitro. It is concluded that embelin probably interferes with reproductive functions in female rats by suppressing ovarian production of sex steroid hormones.     

Role of prostaglandin F2alpha in ovulation.

Using radioimmunoassay procedures, the levels of plasma, uterine and ovarian prostaglandin (PG) F2alpha, and those of plasma estradiol and progesterone were measured in intact, hysterectomized or ovariectomized immature female rats pretreated with PMS and subsequent HCG. Occurrence of ovulation was confirmed at 8 hours after the HCG administration not only in the intact rats but also in the hysterectomzied rats. The levels of plasma estradiol and progesterone, and of uterine and ovarian PGF2alpha rose with the PMS injection alone, but they did not reach the peaks before the HCG administration. Both plasma estradiol and uterine PGF2alpha showed a peak at 2 hours after the HCG injection. These peaks were antecedent 2 or 6 hours before the peaks of ovarian and plasma PGF2alpha, respectively. However, such increase of uterine PGF2alpha does not seem to be indispensable for ovulation, because ovulation could occur in the hysterectomized rats. The levels of ovarian PGF2alpha showed a high plateau from 4 to 8 hours after the HCG injection, and then rapidly decreased after ovulation. The levels of plasma PGF2alpha peaked not only in the intact rats but also in the hysterectomized rats at 8 hours after the HCG treatment. But in the ovariectomized rats, this plasma PGF2alpha peak at 8 hours disappeared and there was no statistical change of plasma PGF2alpha throughout the PMS-HCG treatment. Plasma progesterone gradually increased and reached the maximum at 10 hours after the HCG injection. These results conclude that the main source of increased plasma PGF2alpha during the ovulatory process induced with the PMS-HCG treatment is the ovary, and it is strongly suggested that a rapid increase of PGF2alpha in the ovary may play some important role(s) in the ovulatory process.     

Effect of PGE1 on PGF2 alpha-induced luteolysis in nonbred ewes

Fifty-six ewes were used to study the effects of PGE1 or PGE2 plus PGF2 alpha given into the perivascular space of the ovarian vascular pedicle on luteal function of nonbred ewes. All ewes receiving PGF2 alpha had reduced levels of plasma progesterone and unoccupied receptors for LH at 24 hr after treatment regardless even if they received PGE1 or PGE2 concomitantly. Levels of plasma progesterone in ewes receiving only PGF2 apha were reduced further at 48 hour. Plasma progesterone and unoccupied receptors for LH of ewes receiving PGE2 + PGF2 alpha were maintained at 48 hr at levels seen at 24 hr after treatment, while progesterone in ewes receiving PGE1 + PGF2 alpha at 48 hr returned to levels seen in controls at 48 hr and unoccupied receptors for LH were three fold greater than controls.     

Ovarian capillary blood flow in seasonally anoestrous ewes induced to ovulate by treatment with GnRH

The microsphere technique was used to obtain estimates of ovarian capillary blood flow near ovulation, in 8 seasonally anoestrous ewes, which were induced to ovulate by GnRH therapy. Plasma progesterone concentrations were monitored in jugular blood sampled between Days 4 and 7 after the onset of the preovulatory LH surge. The ewes were then slaughtered. Three of the ewes were treated with a single injection of 20 mg progesterone before GnRH therapy. In these ewes and 1 other, plasma progesterone values increased after ovulation and reached 1.0 ng/ml on Day 7 following the preovulatory LH surge (normal, functional CL), whilst in the other 4 ewes progesterone concentrations increased initially then declined to 0.5 ng/ml by Day 7 (abnormal CL). In the ewes exhibiting normal luteal function, the mean ovarian capillary blood flow was significantly greater (P less than 0.01) than that for ewes having abnormal luteal function. Irrespective of the type of CL produced, capillary blood flow was significantly greater (P less than 0.05) in ovulatory ovaries than in non-ovulatory ovaries. These findings indicate that the rate of capillary blood flow in ovaries near ovulation may be a critical factor in normal development and maturation of preovulatory follicles and function of subsequently formed CL.     

Effects of 2(p-biphenyl) propionic acid on the life span of the corpus luteum of pseudopregnant and cyclic rats

A potent prostaglandin synthesis inhibitor (cyclooxygenase inhibitor), 2(p-biphenyl) propionic acid (BPPA), was administered subcutaneously at the dose of 1 mg twice daily to pseudopregnant and cyclic rats. It was found to prolong the duration of pseudopregnancy by about 10 days and to have little or no effect on estrous cycle length. It did not block ovulation and had little effect on ovulation rate in cyclic rats. BPPA was given to pseudopregnant rats in two trials (one in October-December and the other in March-May) to determine its effect on ovarian weight and plasma progesterone concentration on Days 14, 15 and 16 (Day 0=day of induction of pseudopregnancy). BPPA significantly (P less than 0.001) increased plasma progesterone concentration and reduced ovarian weight. The present data support the hypothesis that prostaglandins cause the normal functional demise of the corpora lutea of pseudopregnancy in the intact rat, and that depressing their synthesis will prolong the functional life span of the corpora lutea.     

Effect of abdominal vagotomy of the pregnant rat on LH and progesterone concentrations and fetal resorption

Abdominal vagotomy on Day 8 of pregnancy in rats decreased the number of live fetuses at Day 16 and increased the number of resorbing fetuses. The activity of delta5-3beta-hydroxysteroid dehydrogenase (3beta-HSD) in the corpus luteum and interstitial gland, LH and progesterone values in plasma and progesterone values in ovarian tissue were all lower in vagotomized rats than in sham-operated controls. Ovarian PGF levels were not affected. We suggest that these effects were caused by a direct effect of vagotomy on LH secretion which in turn lowers 3beta-HSD activity and progesterone levels in ovarian tissue and plasma, leading to fetal resorption.     

Progesterone is not responsible for the blood pressure fall in late-pregnant New Zealand genetically hypertensive rats

In various models of experimental and genetic hypertension in rats, blood pressure is markedly reduced during late pregnancy. The period during which the blood pressure reduction occurs is also the period when plasma progesterone is maximally elevated, and administration of progesterone to renal hypertensive rats has been reported to reduce blood pressure (J. Armstrong, 1959, Proc. Soc. Exp. Biol. Med. 102:452-455). To test the possibility that elevated plasma progesterone is responsible for the blood pressure reduction in late pregnancy, on Day 14 of pregnancy a group of New Zealand genetically hypertensive (NZGH) rats was ovariectomized and implanted with progesterone-filled capsules, to maintain plasma progesterone at low levels just sufficient to maintain pregnancy, and compared with intact, pregnant NZGH. Ovariectomy did not alter the characteristic course of blood pressure reduction seen in late-pregnant intact NZGH rats. In addition, daily administration of progesterone (15 mg/kg, sc) for 14 days did not alter blood pressure of either nonpregnant NZGH rats or New Zealand normotensive rats with chronic 1-kidney, 1-clip hypertension. It is concluded that blood pressure of NZGH rats is reduced to near normotensive levels in late pregnancy, as reported for other models of rat hypertension, but that elevated plasma progesterone levels are not requisite for that reduction and do not reduce blood pressure of renal hypertensive rats.     

Progesterone and the initiation of parturition in the goat

The corpora lutea were surgically removed from 6 goats between 134 and 136 days of pregnancy and progesterone was administered daily. Pregnancy was prolonged past normal term in 4 goats receiving 25 mgs. of progesterone daily as a split dose. However, eventual delivery following progesterone withdrawal was abnormal and foetal mortality high.The progesterone therapy regime maintained maternal jugular plasma concentrations of progesterone in excess of 3 ng/ml. In normal untreated goats, maternal plasma concentrations of progesterone declined over the last 6 days of gestation. In treated goats, plasma concentrations of progesterone fell only after the cessation of therapy. Maternal plasma concentrations of estrogen rose within 24 hours of parturition in normal untreated goats. In the 4 goats in which pregnancy was prolonged, by progesterone administration, maternal plasma concentrations of estrogen were elevated for several days in the period before eventual foetal delivery.     

Pregnancy failure in hypophysectomized rats following LH-RH administration.

Daily LH-RH administration induced a dose related increase in fetal resorption in rats following hypophysectomy on Day 11 or Day 12 of pregnancy. Ovarian and adrenal weights, as well as serum progesterone levels, were significantly decreased by Day 18. Serum progesterone was also significantly lower than control in those animals that remained pregnant after receiving LH-RH post-hypophysectomy. These observations suggest that LH-RH exerts an anti-pregnancy effect via a placental:ovarian axis in this species.     

The effect of frozen ovarian autografts on compensatory ovulation and steroid production in unilaterally ovariectomized rats.

In the present study rats were unilaterally ovariectomized (ULO) and the surgically removed ovary was frozen for 13 days. After allowing the remaining ovary to compensate with respect to number of ova shed, the frozen graft was thawed and transplanted subcutaneously to determine the effect on ovulation number, cycle length, uterine weight, ovarian weight and plasma levels of estradiol-17beta (E2) and progesterone. Rats ULO at 45 days of age, which received an autograft 13 days later, had a decrease in the number of eggs shed as compared to control ULO rats (6.4 +/- 0.8 vs. 11.1 +/- 0.9 eggs, respectively) and a decrease in plasma E2 (14.5 +/- 1.7 VS. 21.0 +/- 1.5 PG/ML, respectively). No differences were observed in progesterone concentration, uterine weight, ovarian weight or cycle length. In contrast, rats ULO at 31 days of age, which received an autograft 13 days later, showed no differences in comparison to control ULO rats. Castrates which received ovarian autografts developed cycling vaginal smears and had increased E2 (31.9 +/- 4.3 pg/ml) and decreased progesterone (18.3 +/- 1.9 ng/ml) levels. Since ULO animals with autografts shed fewer ova, the present study demonstrates that the amount of ovarian tissue influences ovulation number either by utilization of gonadotropins or by an, as yet, undefined mechanism.     

Stimulation of ovarian cyclic guanosine 3',5'-monophosphate levels by the subunits of human chorionic gonadotropin

Injection of human chorionic gonadotropin (HCG) into the tail vein of superovulated rats resulted in a significant (P<0.01) increase in peripheral plasma progesterone without a concomitant increase in ovarian cyclic GMP (cGMP) levels. However, when equimolar quantities of α and β subunits of HCG were injected, a significant increase in plasma progesterone was accompanied by a concomitant and significant (P<0.01) increase in ovarian cGMP levels. The observation that these subunits increase ovarian cGMP levels without increasing cAMP suggests the possibility of cGMP involvement in steroidogenesis induced by subunits.     

Plasma progesterone concentration depends on sampling site in pigs

The objective of this study was to examine a possible difference in progesterone concentrations between the systemic venous blood and the caudal vena cava in early pregnant gilts. Nineteen crossbred pregnant gilts were offered three different regimens of feeding to examine influence of feeding on the secretion pattern of progesterone. The groups were high (H-H), low (L-L) and low-high (L-H) receiving 3.6, 1.8 and 1.8/3.6 kg/day, respectively. Catheters were placed in a jugular vein and the caudal vena cava (to sample ovarian secretion) on day 19 of pregnancy. Two consecutive samples taken at 30-min intervals were collected four times a day for 5 days (days 20-24). In addition, three gilts were simultaneously sampled from both catheters at 30-min intervals for 12 h on day 22. Progesterone concentration was significantly lower in the jugular vein compared with the caudal vena cava in all three feeding groups (P<0.001). An indication of episodic pattern of progesterone production occurred in plasma collected from the caudal vena cava, but not from the jugular vein. Dietary intake did not cause a profound effect on plasma progesterone concentrations during days 20-24 of gestation. It seemed that ovarian progesterone was released into the vena cava in an episodic pattern and there were implications that these episodes were temporally associated with LH pulses.     

Ovarian and adrenal function during the estrous cycle of the sow.

The concentrations of plasma estrogens, progesterone, and corticosteroids and of urinary pregnanediol, pregnanetriol, ketogenic steroids, and corticosteroids were determined as indicators of ovarian and adrenal function throughout a normal sow's estrous cycle. Two broad peaks of plasma estrogen, one lasting 11–12 days during estrus and another 6-day peak period during the early part of the luteal phase were detected. Plasma progesterone was elevated during the late follicular and luteal phase. Two broad peaks of plasma corticoids appeared, one following the decrease of plasma progesterone and the second 7–14 days later. Those elevations in plasma corticoids occurred when estrogen titres were elevated. Urinary determinations generally reflected plasma findings. Estrogen levels began to rise during the follicular phase while a reasonably high progesterone level was evident. Estrogen titres never decreased to non-detectable levels. An interrelationship between adrenal function and ovarian estrogen production is suggested.     

Involvement of the corticoadrenal hormones in the pheromonal restoration of reproductive activity in aging rats

The involvement of the adrenal progesterone and corticosterone in the early gonadotropin secretion associated with the pheromonal restoration of ovarian cyclic activity (PRCA) in aging female rats is studied. PRCA is induced by male urinary pheromones and is preceded by an alpha-adrenergic-mediated release of the hypothalamic decapeptide luteinizing hormone-releasing hormone and plasma increases of estradiol, progesterone and the gonadotropins luteinizing hormone and follicle stimulating hormone. Aging reproductive Wistar female rats were used to study the effects of bilateral adrenalectomy and of a subcutaneous injection of the antisteroid RU486 on plasma levels of corticosterone, progesterone and gonadotropins in rats stimulated with nasal spraying of male urine (MU) or saline. The results demonstrate that progesterone and corticosterone released by MU are from adrenal origin, and that these adrenal secretory products are critical for MU-induced increase of gonadotropins. This suggests that olfactory stimulation of ACTH release stimulates adrenal release of progesterone and corticosterone, and both trigger the events that initiate the activation of the hypothalamus-pituitary-ovarian axis that leads to PRCA.     

Release of ovarian progesterone during the rat oestrous cycle by ganglionic cholinergic influence: the role of norepinephrine

The coeliac ganglion neurons, whose axons constitute the superior ovarian nerve (SON), contain cholinergic receptors. The aim of this work was to study the effect of cholinergic agents added to the coeliac ganglion on the release of ovarian progesterone in the coeliac ganglion-SON-ovary in vitro system. We also analyzed the release of norepinephrine in the ovarian compartment and its possible relationship with the release of progesterone. After the addition of cholinergic agents in the ganglion compartment, progesterone release was determined by radioimmuneassay (RIA) and norepinephrine by catecholamine assay (HPLC). The release of progesterone and norepinephrine in the ovary compartment was studied during period of 180 min in pre-oestrus (PE), oestrus (E), dioestrus day 1 (D1) and dioestrus day 2 (D2) rats. The most relevant results concerning the action of acetylcholine were found on PE and dioestrus. On PE, the pre-ovulatory peak of progesterone, which is known to respond to the endocrine action, was not modified by neural effect of acetylcholine in our scheme. On the other hand, the progesterone peak occurs in the afternoon of D1, which has been described as independent of the gonadotrophic action but was inhibited by neural effect of acetylcholine in our experimental scheme. This action on D1 was accompanied by a decrease of norepinephrine release in the ovary compartment. We conclude that the action of cholinergic agents varies according to the oestrous cycle stage and constitutes one of the factors governing the secretory activity of the ovarian steroids, in this case, progesterone.     

Functional and morphological changes in the hypothalamo-pituitary-gonadal axis of aged female rats.

Age-related functional and morphological alterations in the hypothalamo-pituitary-gonadal axis were investigated in old recurrently pseudopregnant (RPP) female rats, and these alterations were compared with those in young diestrous rats. LHRH in the median eminence (ME) and mediobasal hypothalamus (MBH) as well as plasma FSH, LH, and progesterone were measured by RIA. LHRH in the lateral ME (LME) and pituitary FSH and LH were evaluated by morphometry and densitometrical immunocytochemistry. Furthermore, by light microscopy, we classified and counted the number of ovarian follicles and corpora lutea. LHRH concentrations in the ME and MBH were similar in old and young rats, whereas in old rats, plasma FSH was markedly increased, LH was moderately increased, and plasma progesterone was unchanged. The number and the total area and immunoreactivity of LHRH-labeled axon cross sections in the LME were reduced in old rats. The number of nucleated FSH-labeled cells and total FSH area and immunoreactivity were almost twice in old compared with young animals. The measurements of LH-labeled cells were not different between the two groups. In old rats, the numbers of ovarian follicles and corpora lutea were reduced and that of atretic follicles increased. In conclusion, age-related morphological impairments of LHRH axons associated with an increased number of FSH gonadotropes and higher plasma FSH in our old RPP rats suggest hypothalamic and pituitary disturbances, which may largely contribute to the complex hormonal disarrangement responsible for the decline of reproductive functions in old female rats.