Prostaglandins involvement in the formation of luteinized unrupted follicles in the cyclic female rat

The purpose of this investigation was to study the role played by prostaglandins in advanced ovulation and in the formation of luteinized unrupted follicles (LUF) in cyclic female rats. Dose related effects on ovulation were observed in rats given LH on diestrus 2 at 16.30. A significant positive correlation was observed between the number of postovulatory corpura lutea (POCL) and the increasing doses of LH. By contrast the number of LUF was negatively correlated with LH. Indomethacin treatment by 6h30 after administration of an ovulatory LH dose significantly increased the occurence of LUF at the expense of POCL. Conversely PGF_2α when admiststered by 6h30 after a subovulatory LH stimulation enhanced in a dose dependent manner the number of POCL with respect to the LH treated controls. Under a similar treatment with a subovulatory dose of LH, PGE₂ remained without ovulatory effects. The mechanisms of the formation of LUF are discussed on the basis of these results.     

Olfactory environment and coitus-induced ovulation and/or luteinization in the cyclic female rat

The role of pheromones in the process of ovulation and/or luteinization induced by coitus was studied in female rats primed with estradiol benzoate (EB) and early mated during 4-day cycles. The effects of coitus were evaluated by using 1) the proportion of females displaying postovulatory corpora lutea (POCL) and/or luteinized unruptured follicles (LUF), and 2) the ovulation coefficient (OC) computed in each female by dividing the number of POCL by the total number of POCL and LUF. A greater proportion of females displayed ovulation and/or luteinization following coitus than those given EB only. This proportion was slightly but significantly increased when females were exposed to bedding soiled with 5 ml of male urine prior to mating, an effect prevented by complete olfactory bulbectomy. However, in the females given EB only, the pheromonal stimulus remained inefficient. Weak ovulatory effects of coitus, as assessed by low OC values, were noted whether the females were exposed or not to male urine. An increase in the frequency of lordosis occurred after olfactory bulbectomy, but no significant changes in OC values were observed in this experimental group. It was concluded that pheromones, act as a primer on the neuroendocrine mechanisms governing ovulation in the cyclic female rat, as in other species of rodents.     

Progesterone secretion by luteinized unruptured follicles in mature female rats

The ability of luteinized unruptured follicles (LUF) to display luteal activity was investigated in mature female rats. Previous findings in our laboratory showed that increasing doses of LH, when injected on late diestrus in 4-day cyclic rats, were capable of inducing the formation of either LUF or postovulatory corpora lutea (POCL) in a dose dependent manner. Four-day cyclers were injected on diestrus 2 at 4.30 p.m. (day 0) with 2.7 micrograms or 5.4 micrograms/100 g of an ovine LH preparation (x 2.94 NIH LH S3) and were killed at different times during the three successive days following injection. Natural 4-day cyclers were killed at corresponding times following spontaneous LH release on proestrus afternoon (day 0). Both LUF and POCL were observed in LH-treated females. LUF appeared more numerous in females given 2.7 micrograms LH than in both natural cyclers and in females injected with 5.4 micrograms LH. On day 1 during the rising phase of luteal activity serum progesterone (P) level did not differ in the three groups despite the high number of LUF in females given 2.7 micrograms LH. On day 2 at 11 a.m. lower P values were observed in both groups of LH-treated females than in natural cyclers, this corresponding to a greater proportion of LUF in the former than in the latter. On day 2, at 5 p.m. by the time of full activity of POCL in natural cyclers, P did not differ in the three groups irrespective of the relative number of ruptured or unruptured follicles. On day 3, P sharply declined in LH-treated and natural cyclers. These results suggest that LUF are capable of secreting P during a period corresponding to the duration of corpus luteum's life span in cyclic female rats.     

Follicle and systemic hormone interrelationships during induction of luteinized unruptured follicles with a prostaglandin inhibitor in mares

The objective was to determine differences in follicle and reproductive hormone characteristics in mares with ovulatory and flunixin meglumine (FM)-induced anovulatory cycles. Estrous mares were given 1500 IU hCG when the follicle was ≥ 32 mm (0 h). In Experiment 1, control mares (n = 7) were not treated further. The remaining mares (n = 11) were given 1.7 mg/kg FM i.v. twice daily, from 0 to 36 h after hCG treatment. Blood samples and ultrasonographic examinations were performed every 12 h. All control mares ovulated normally between 36 and 48 h. In contrast, eight of 11 FM mares did not ovulate, but developed luteinized unruptured follicles (LUFs). Three FM-treated mares did not develop conventional LUFs. Plasma progesterone concentrations were lower (P < 0.05) in LUF mares at 96, 120, and 216 h than in controls, whereas plasma LH concentrations were higher (P < 0.05) between 108 and 120 h in LUF mares than in controls. Plasma concentrations of PGFM and estradiol did not differ significantly between groups. In Experiment 2, the three mares that did not develop LUFs were treated, during the consecutive cycle, with the same dose of FM but with increased frequency at zero, 12, 24, 30, 36, and 48 h after hCG. One mare formed a LUF, whereas the other two did not. These two mares had lower LH concentrations than LUF or control mares in the two consecutive cycles. In conclusion, systemic treatment with FM blocked ovulation in 73% of treated mares. Mares with LUFs had lower progesterone and higher LH concentrations than control mares.     

A hormonal and temporal analysis of the mechanisms involved in the control of ovulation induced by hemiovariectomy in the rat

The present study was undertaken to investigate the mechanisms of the stress-related ovulatory effects of hemicastration in the rat. Previous work (Roos et al., 1976) had shown that ovulation induced by unilateral ovariectomy (ULO) was suppressed in adrenalectomized females when ULO was performed on dioestrus III at 10--11 h in 5-day cyclic rats. Using the same experimental schema an increase in blood progesterone within 1 to 4 hours after ULO has been found to be present in adrenal intact females and suppressed in adrenalectomized rats. PB treatment (30 mg/kg, i.p.) concomitant with ULO at 10--11 h on dioestrus III significantly decreased the number of ovulating females without preventing blood progesterone concentration to increase at 12--13 h. A partial blockade of ovulation resulted from PB injection at 13 or 18 h. The ovulatory effects of ULO observed in females injected with PB at 23 h on dioestrus III or at 5 h on prooestrus were identical to those observed in hemiovariectomized non PB treated females. Only a small proportion of hemiovariectomized females displayed an LH release at 15--16 h and 17.30 h--18.30 h on dioestrus III. In contrast a significant FSH release was observed in this interval of time following ULO. Microscopic examination of the ovaries on prooestrus at either 11 h or 16 h revealed the presence of corpora lutea with morphological features corresponding to very different stages of development. We can conclude that progesterone of adrenal origin constituted the trigger of ovulation and caused LH-release during a time period extending from 13 h to 23 h on dioestrus III following ULO in the rat.     

Supplementation of equine early spring transitional follicles with luteinizing hormone stimulates follicle growth but does not restore steroidogenic activity

This study was conducted to test the hypothesis that supplementation of growing follicles with LH during the early spring transitional period would promote the development of steroidogenically active, dominant follicles with the ability to respond to an ovulatory dose of hCG. Mares during early transition were randomly assigned to receive a subovulatory dose of equine LH (in the form of a purified equine pituitary fraction) or saline (transitional control; n = 7 mares per group) following ablation of all follicles >15 mm. Treatments were administered intravenously every 12 h from the day the largest follicle of the post-ablation wave reached 20 mm until a follicle reached >32 mm, when an ovulatory dose of hCG (3000 IU) was given. Saline-treated mares during June and July were used as ovulatory controls. In a preliminary study, injection of this pituitary fraction (eLH) to anestrus mares was followed by an increase in circulating levels of LH (P < 0.01) but not FSH (P > 0.6). Administration of eLH during early transition stimulated the growth of the dominant follicle (Group x Day, P < 0.00001), which attained diameters similar to the dominant follicle in ovulatory controls (P > 0.1). In contrast, eLH had no effect on the diameter of the largest subordinate follicle or the number of follicles >10 mm during treatment (P > 0.3). The numbers of mares that ovulated in response to hCG in transitional control, transitional eLH and ovulatory control groups (2 of 2, 3 of 5 and 7 of 7, respectively) were not significantly different (P > 0.1). However, after hCG-induced ovulation, all transitional mares returned to an anovulatory state. Circulating estradiol levels increased during the experimental period in ovulatory controls but not in transitional eLH or transitional control groups (Group x Day, P = 0.013). In addition, although progesterone levels increased after ovulation in transitional control and transitional eLH groups, levels in these two groups were lower than in the ovulatory control group after ovulation (Group, P = 0.045). In conclusion, although LH supplementation of early transitional waves beginning after the largest follicle reached 20 mm promoted growth of ovulatory-size follicles, these follicles were developmentally deficient as indicated by their reduced steroidogenic activity.     

Role of the double luteinizing hormone peak, luteinizing follicles, and the secretion of inhibin for dominant follicle selection in Asian elephants (Elephas maximus)

Elephants express two luteinizing hormone (LH) peaks timed 3 wk apart during the follicular phase. This is in marked contrast with the classic mammalian estrous cycle model with its single, ovulation-inducing LH peak. It is not clear why ovulation and a rise in progesterone only occur after the second LH peak in elephants. However, by combining ovarian ultrasound and hormone measurements in five Asian elephants (Elephas maximus), we have found a novel strategy for dominant follicle selection and luteal tissue accumulation. Two distinct waves of follicles develop during the follicular phase, each of which is terminated by an LH peak. At the first (anovulatory) LH surge, the largest follicles measure between 10 and 19.0 mm. At 7 ± 2.4 days before the second (ovulatory) LH surge, luteinization of these large follicles occurs. Simultaneously with luteinized follicle (LUF) formation, immunoreactive (ir) inhibin concentrations rise and stay elevated for 41.8 ± 5.8 days after ovulation and the subsequent rise in progesterone. We have found a significant relationship between LUF diameter and serum ir-inhibin level (r(2) = 0.82, P < 0.001). The results indicate that circulating ir-inhibin concentrations are derived from the luteinized granulosa cells of LUFs. Therefore, it appears that the development of LUFs is a precondition for inhibin secretion, which in turn impacts the selection of the ovulatory follicle. Only now, a single dominant follicle may deviate from the second follicular wave and ovulate after the second LH peak. Thus, elephants have evolved a different strategy for corpus luteum formation and selection of the ovulatory follicle as compared with other mammals.     

A dopamine agonist stimulates progesterone secretion from adrenal gland

Administration of either progesterone (P) or a dopamine agonist, Legrotrile mesylate (LM), have been shown to induce the ovulatory release of LH in rats. In order to elucidate the mode of action of dopamine agonists we studied the effects of LM on P secretion by the adrenals. A subcutaneous injection of LM, in doses which induce ovulation, stimulated adrenal P secretion in ovariectomized, estrogen-primed rats and in castrate male rats. Peak plasma P concentrations were found at 2 h with a gradual return to pre-injection levels at 6 hrs following LM injection. These results raise the possibility that P increments following LM administration may be responsible for inducing ovulation in young and old rats.There is considerable evidence to show that modification of hypothalamic monoamine metabolism by pharmacologic agents results in markedly altered pituitary gonadotropin secretion [1]. Lergotrile mesylate (LM), a dopamine agonist, has been shown to advance the ovulatory release of LH on proestrus and induce ovulation and cyclicity in anovulatory aged rats [2,3]. Everett [4] observed that progesterone (P) administration induced ovulation in rats rendered anovulatory under constant light and also advanced ovulation by one day if administered to 5-day cycling rats. A similar effect of LM on ovulation advancement has been noted (Clemens, personal communication). These similarities in action of P and LM on ovulation in young and aged rats, led us to speculate that LM may stimulate P secretion which in turn may elicit the discharge of ovulatory hormones. To test this hypothesis we examined the effects of LM on progesterone secretion in gonadectomized rats.     

Ovarian and endocrine responses in the cat after coitus

LH release leading to ovulation was induced in 17 of 29 oestrous periods. The time of ovulation after coitus was determined by histological examination or by observation at laparotomy of ovaries in situ. Histological methods revealed that ovulation was complete in most follicles (9 of 13) at 32 h post coitum and in all follicles that were involved in the ovulatory process by 36 h. When laparotomy was used, no signs of preovulatory change were noted at the first observation time, 22 h post coitum, but in 4 cycles in which the entire process of ovulation was observed, the ovulatory process occurred between 23 and 28 h (3 follicles), 23 and 27 h (2 follicles), 25 and 28 h (3 follicles), and 25 and 29 h (3 follicles) post coitum. The first ovulatory process noted was complete at 25 h post coitum. In cats, LH release continued over a 16-h period before returning to baseline (long surge), values being 616 +/- 180 ng/ml at 1/2 h and 941 +/- 154 ng/ml at 2 h post coitum. In 6 cats the LH release pattern was limited to a 4-h period (short surge), values being 537 +/- 218 ng/ml at 1/2 h and 353 +/- 245 ng/ml plasma at 2 h and basal (49 +/- 18 ng/ml) by 4 h post coitum. Decreased secretion of oestrogen by follicles in animals undergoing ovulation was first observed at 16 h post coitum. It is concluded that coitus induces LH release within minutes in the cat and that ovulation begins about 24 h later and finishes by about 32 h post coitum. Only one coital input can cause LH release for as long as 16-20 h although shorter periods of LH release (4 h or less) can result in ovulation.     

Effects of aromatizable and nonaromatizable androgen treatments on luteinizing hormone receptors and ovulation induction in immature rats

The effects of androgen pretreatment on follicle-stimulating hormone (FSH)-stimulated luteinizing hormone (LH) receptor induction in ovarian granulosa cells was examined. Immature female rats were treated with various doses (0.1-5 mg/rat) of testosterone (T), 5 alpha-dihydrotestosterone (DHT), 5 alpha-androstane-3 alpha,17 beta-diol (3 alpha-diol), or 5 alpha-androstane-3 beta,17 beta-diol (3 beta-diol). Subsequent follicular development was stimulated by treatment with ovine FSH. LH receptor induction in granulosa cells and ovulatory responses to 10 IU human chorionic gonadotropin (hCG) were examined. Since LH receptor induction requires the synergistic action of both FSH and estradiol, the effects of the androgen pretreatment on FSH-stimulated estradiol production were also examined. Dihydrotestosterone treatment at doses greater than 1 mg inhibited LH receptor induction by approximately 70%, which resulted in absent ovulatory responses. Treatment with 1 mg or more of T or 3 alpha-diol had no effect on LH receptor induction, yet the hCG-stimulated ovulation rate was reduced to 40% of that seen in vehicle-treated controls. 3 beta-Diol, at a dose of 1 mg/rat, did not affect LH receptor induction but did reduce hCG-stimulated ovulation responses. No significant effects of androgen treatment on ovarian or uterine weight or FSH-stimulated estradiol production were observed. These results suggest that androgens can act at multiple sites to inhibit ovarian follicular development and function. In addition these studies demonstrate that, although LH receptor induction is necessary, it may not be a sufficient condition to ensure ovulation of ovarian follicles.     

A possible dual mechanism of the anovulatory action of antiprogesterone RU486 in the rat

The purpose of these experiments was to investigate the mechanism of the anovulatory action of antiprogesterone RU486 (RU486) in rats by studying its effects on follicular growth, secretion of gonadotropins and ovarian steroids, and ovulation. Rats with 4-day estrous cycles received injections (s.c.) of either 0.2 ml oil or 0.1, 1, or 5 mg of RU486 at 0800 and 1600 h on metestrus, diestrus, and proestrus. At the same times, they were bled by jugular venipuncture to determine serum concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH), 17 beta-estradiol (E), and progesterone (P). On the morning of the day after proestrus, ovulation and histological features of the ovary were recorded. Rats from each group were killed on each day of ovarian cycle to assess follicular development. Rats treated similarly were decapitated at the time of the ovulatory LH surge and blood was collected to measure LH. The serum levels of LH increased and those of FSH decreased during diestrus in rats treated with RU486. Neither E nor P levels differed among the groups. Treatment with RU486 caused both a blockade of the ovulation and an increase in ovarian weight in a dose-dependent manner. At the time of the autopsy (the expected day of ovulation), rats treated with 1 mg RU486 had ovaries presenting both normal and post-ovulatory follicles and unruptured luteinized follicles. Rats treated with 5 mg RU486 presented post-ovulatory follicles without signs of luteinization. The number of follicles undergoing atresia increased in rats treated with RU486. Rats treated with 5 mg RU486 exhibited a significant decrease in ovulatory LH release. The mechanism by which RU486 produces the ovulatory impairment in rats seems to be dual: first, by inducing inadequate follicular development at the time of the LH surge and second, by reducing the amount of ovulatory LH released. The physiological events-decreased basal FSH secretion and follicular atresia-that result from use of RU486 cannot be elucidated from these experiments and should be investigated further.     

Delta-9-tetrahydrocannabinol attenuates luteinizing hormone release induced by electrochemical stimulation of the medial preoptic area.

Despite diverse pharmacological actions, drugs commonly used for blocking ovulation in the rat have not been observed to exert differential effects on the LH response to preoptic stimulation, thus suggesting blocking action above the final hypothalamic GnRH pathway. To determine if ovulatory blockade by delta-9-tetrahydrocannabinol (THC) is consistent with that scheme, LH surges evoked by preoptic stimulation were contrasted with those elicited during blockade by atropine (ATR), a classic ovulation-blocking agent with which other drugs have been compared. THC (10 mg/kg) or ATR (350 mg/kg) treatment before the proestrous critical period uniformly blocked LH release and ovulation in sham-stimulated rats. Preoptic stimulation evoked LH surges after both drug treatments (p less than 0.001), peak levels increasing with the intensity of stimulation (p less than 0.05). However, both maximum LH concentration (p less than 0.05) and total integrated LH release (p less than 0.01) were lower in THC-blocked rats. Inspection of the oviducts revealed no difference in the incidence of ovulation or the number of ova discharged. The reduced LH response during THC blockade was not attributable to variation in the extent or locus of histologically determined stimulation sites. These results distinguish THC from ATR and, by extension, other blocking drugs that do not overtly affect the LH response to preoptic stimulation. Thus, ovulatory blockade by THC may involve a different mechanism, which likely includes inhibitory action within the preoptic-to-tuberal GnRH pathway.     

Effects of inhibitors of protein synthesis on the ovulatory process of the perfused rat ovary

The effects of two different protein synthesis inhibitors (cycloheximide and puromycin) on the ovulatory process were examined in vitro using a perfused rat ovary model. Ovaries of PMSG (20 i.u.)-primed rats were perfused for 21 h. Release of cyclic adenosine 3',5'-monophosphate (cAMP) and steroids (progesterone, testosterone, and oestradiol) was measured and the number of ovulations was estimated by counting released oocytes. Unstimulated control ovaries did not ovulate whereas addition of LH (0.1 microgram/ml) plus 3-isobutyl-1-methylxanthine (IBMX; 0.2 mM) resulted in 16.7 +/- 3.5 ovulations per treated ovary. Cycloheximide (5 micrograms/ml) totally inhibited the ovulatory effect of LH + IBMX when present from the beginning of the perfusions and also when added 8 h after LH + IBMX. No inhibition was seen when cycloheximide was added 10 h after LH + IBMX (1-1.5 h before the first ovulation; 15.2 +/- 4.4 ovulations per treated ovary). Puromycin (200 micrograms/ml) completely blocked ovulation when present from the beginning of the perfusions and the inhibition was congruent to 60% (6.5 +/- 2.2 ovulations per treated ovary) when the compound was added 8 h after LH + IBMX. Both inhibitors increased LH + IBMX-stimulated cAMP release substantially, but decreased the release of progesterone, testosterone and oestradiol. These results indicate that de-novo protein synthesis is important late in the ovulatory process for follicular rupture to occur.     

Time-dependent ovulation inhibition of a selective progesterone-receptor antagonist (Org 31710) and effects on ovulatory mediators in the in vitro perfused rat ovary

Progesterone (P) is one of several local mediators in the ovulatory cascade in the rat. The precise mechanisms of action for P in ovulation and in what phase of the ovulatory process P is critical, however, need to be clarified. The present study used a selective P-receptor antagonist, Org 31710, in the in vitro perfused rat ovary model to examine the local role of P and possible effects on prostaglandin (PG) and plasminogen-activator (PA) release in ovulation. Ovaries from eCG (15 IU)-primed rats were perfused for 20 h with LH (0.2 microg/ml) and 3-isobutyl-1-methylxanthine (IBMX, 200 microM) to induce ovulation (median = 10.0, 25%-75% range = 8.5-13). Org 31710 was added at either 0, 3.5, 7, or 9 h after LH+IBMX, resulting in significant suppression of ovulation after addition at 0 and 3.5 h (1.0, 1-5.5; and 5.0, 2.5-7.75 ovulations, respectively) but no suppressive effect when added at later time points. Progesterone and estradiol levels in the perfusion media were increased after LH+IBMX but were not affected by the presence of Org 31710. Ovarian tissue levels of PGE(2), PGF(2 alpha), and PA activity were measured in ovaries that had been perfused for 10 h, a time that was 2 to 5 h before anticipated ovulation. The presence of Org 31710 significantly decreased the levels of PGE(2), PGF(2 alpha), and PA activity. These results suggest that P is essential in ovulation during the initial stages of the ovulatory process. The effect of P to facilitate ovulation seems to relate to stimulation of the PG- and PA-mediator systems.     

Follicular deviation and acquisition of ovulatory capacity in bovine follicles.

Selection of dominant follicles in cattle is associated with a deviation in growth rate between the dominant and largest subordinate follicle of a wave (diameter deviation). To determine whether acquisition of ovulatory capacity is temporally associated with diameter deviation, cows were challenged with purified LH at known times after a GnRH-induced LH surge (experiment 1) or at known follicular diameters (experiments 2 and 3). A 4-mg dose of LH induced ovulation in all cows when the largest follicle was > or =12 mm (16 of 16), in 17% (1 of 6) when it was 11 mm, and no ovulation when it was < or =10 mm (0 of 19). To determine the effect of LH dose on ovulatory capacity, follicular dynamics were monitored every 12 h, and cows received either 4 or 24 mg of LH when the largest follicle first achieved 10 mm in diameter (experiment 2). The proportion of cows ovulating was greater (P < 0.05) for the 24-mg (9 of 13; 69.2%) compared with the 4-mg (1 of 13; 7.7%) LH dose. To determine the effect of a higher LH dose on follicles near diameter deviation, follicular dynamics were monitored every 8 h, and cows received 40 mg of LH when the largest follicle first achieved 7.0, 8.5, or 10.0 mm (experiment 3). No cows with a follicle of 7 mm (0 of 9) or 8.5 mm (0 of 9) ovulated, compared with 80% (8 of 10) of cows with 10-mm follicles. Thus, follicles acquired ovulatory capacity at about 10 mm, corresponding to about 1 day after the start of follicular deviation, but they required a greater LH dose to induce ovulation compared with larger follicles. We speculate that acquisition of ovulatory capacity may involve an increased expression of LH receptors on granulosa cells of the dominant follicle and that this change may also be important for further growth of the dominant follicle.     

Qualitative and quantitative data on experimental luteinization in the female rat

The dose-effect of 1.5-16 mcg luteinizing hormone (LH) per 100 gm body weight injected in rats at 1100-1200 on proestrus was compared with 30 mg meprobamate given to controls at the same time, on luteinization and ovulation seen in serial ovarian sections. The WII Wistar rats were killed. Luteinization with or without ovulation increased with dose (1.5, 3, 5, 8, and 16 mcg) of LH to a plateau (90%) above the 5 mcg dose, compared with 18% in controls. 2-5 animals in each dose group had preluteinized follicles, characterized by a dissociation of the cumulus oophorus from the granulosa. The absolute frequency of ovulation increased linearly with LH dose, but the frequency of ovulation among rats that luteinized was invariant. The coefficient of ovulation, calculated as the mean incidence of ovulation in relation to the total number of luteinized or preluteinized follicles in each rat, decreased from .769 in controls to .580 in the 3 mcg group, then rose to .916 in the 16 mcg group. Thus, in proestrous rats, low doses of LH induce corpora lutea with retained ova. The threshold dose of LH for luteinization and for ovulation is lower in proestrus than in diestrus II, but varies slightly in different strains of rats.     

Ability of progesterone to reverse anti-androgen (hydroxyflutamide)-induced interference with the preovulatory LH surge and ovulation in PMSG-primed immature rats

In Exp. 1, PMSG was injected to 26-day-old prepubertal rats to induce ovulations. On Day 2 (2 days later, the equivalent of the day of pro-oestrus) they received at 08:00 h 5 mg hydroxyflutamide or vehicle and at 12:00 h 2 mg progesterone or testosterone or vehicle. Animals were killed at 18:00 h on Day 2 or at 09:00 h on Day 3. Progesterone but not testosterone restored the preovulatory LH surge and ovulation in hydroxyflutamide-treated rats. In Exp. 2, 2 mg progesterone or testosterone were injected between 10:30 and 11:00 h on Day 2, to advance the pro-oestrous LH surge and ovulation in PMSG-primed prepubertal rats. Injection of hydroxyflutamide abolished the ability of progesterone to advance the LH surge or ovulation. Testosterone did not induce the advancement of LH surge or ovulation. In Exp. 3, ovariectomized prepubertal rats implanted with oestradiol-17 beta showed significantly (P less than 0.01) elevated serum LH concentrations at 18:00 h over those observed at 10:00 h. Progesterone injection to these animals further elevated the serum LH concentrations at 18:00 h, in a dose-dependent manner, with maximal values resulting from 1 mg progesterone. Hydroxyflutamide treatment significantly (P less than 0.003) reduced the serum LH values in rats receiving 0-1 mg progesterone but 2 mg progesterone were able to overcome this inhibition. It is concluded that progesterone but not testosterone can reverse the effects of hydroxyflutamide on the preovulatory LH surge and ovulation. It appears that hydroxyflutamide may interfere with progesterone action in induction of the LH surge, suggesting a hitherto undescribed anti-progestagenic action of hydroxyflutamide.     

Luteinizing hormone requirements for ovulation in the rat.

Luteinizing hormone requirements for ovulation induction were studied in proestrous rats through detailed observation of the preovulatory surge, through various forms of LH injection under sodium pentobarbital blockade, and through estimation of LH uptake by the ovary. Blood LH levels in individual proestrous rats were obtained every 30 min and grouped according to their peak time (designated 0 h); mean LH levels higher than 7 and 5 ng/ml continued for 30 min and 2.5 h, respectively, the pituitary LH contents at 1400 and 2000 h on the day of proestrus were 2.1 and 0.7 micrograms, respectively, indicating that the amount of LH secreted during the surge was at least 1.4 micrograms. Single intravenous injections of 2 micrograms and 1 micrograms of pure rat LH (NIDDK-rLH-I-7; FSH and prolactin contaminations: 0.02% and less than 0.01%, respectively) to sodium pentobarbital-blocked rats induced ovulation in 4 out of 4 rats and 4 out of 6 rats, respectively, while 500 ng failed to induce ovulation in any (out of 7) rats. Two injections of 300 ng each with an interval of 20 min induced ovulation in 3 out of 8 rats, but if the interval was prolonged to between 30 and 120 min, 100% ovulation was obtained. Blood LH levels in these experiments indicated that a lower long-lasting LH level (about 5 ng/ml blood) is more important than a short, high level for ovulation induction. It was also shown that this level of LH could be given in separate doses if the interval was 30-120 min.(ABSTRACT TRUNCATED AT 250 WORDS)     

Inhibition of ovulation in the rat by electrical stimulation of the lateral amygdala.

Effects of electrical stimulation of the lateral amygdaloid areas on ovulatory gonadotropin release were examined in adult Wistar female rats. Electrical stimulation was applied in rats in proestrus under ether anesthesia with square wave pulses of 0.5 ms duration and 100 Hz frequency for 30 min (30 sec on and off). Stimulation of the lateral amygdala blocked ovulation in 50% of animals when it was applied between 13:30 and 14:30 with a current of 300 muA. Stimulation of the medial amygdala under the same experimental condition was absolutely ineffective to block ovulation. Sham stimulation was also ineffective. In determining the gonadotropin concentration in serum, the stimulation into the lateral amygdala was observed to inhibit the ovulatory release of LH, FSH and prolactin. It may be said that the lateral amygdaloid area participates in the control of gonadotropin release in an inhibitory manner.     

Ovulation inhibition in the pregnant mare's serum gonadotropin-treated immature rat: a bioassay for luteinizing hormone-releasing hormone antagonists

A convenient method for evaluating the biological activity of luteinizing hormone-releasing hormone (LHRH) antagonists was devised. Pregnant mare's serum gonadotropin (PMSG) treatment of immature rats is known to stimulate follicular growth and estrogen production, that in turn stimulates the release of LHRH which triggers an ovulatory discharge of luteinizing hormone (LH) from the pituitary. The present bioassay of the antagonists is based on the inhibition of ovulation in the PMSG-treated rats. Twenty-eight-day-old Sprague Dawley rats maintained under a light period of 12 h/day (lights on at 0630 h) were given 10 IU of PMSG s.c. at 0930 h. On Day 30 of age the antagonist was given s.c. at 1430 h. The rats were killed on the following morning and the oviducts examined for the presence of ova. In addition, the antagonists were compared in their ability to inhibit serum testosterone levels in adult male rats. In the PMSG-treated rats the order of ovulation-inhibiting potency of the following antagonists was: [Ac-D-NAL(2)1,4FD-Phe2,D-Trp3,D-Arg6]-LHRH (LHRH-1) greater than [Ac-delta 3 Pro1,4FD-Phe2,D-NAL(2)3.6]-LHRH (LHRH-2) greater than [Ac-delta 3 Pro1,4FD-Phe2,D-Trp3,6]-LHRH (LHRH-3). The order of potency was confirmed by their antitesticular effects in adult male rats.