Plasma concentrations of inhibin a and follicle-stimulating hormone differ between cows with two or three waves of ovarian follicular development in a single estrous cycle

Patterns of ovarian follicle development were monitored daily in Holstein-Friesian cows that had two (n = 4) or three (n = 4) waves of ovarian follicle development during a single estrous cycle. The plasma from daily blood samples was used in assays for inhibin A, FSH, progesterone, and estradiol-17beta. Mean cycle lengths for cows with two and three waves were 21.8 and 25.3 days, respectively (P < 0.02). Although the average number of follicles >3-mm diameter on each pair of ovaries was similar for two- and three-wave cows on Days 2, 3, and 4 (Day 0 = day of ovulation; 8.6 vs. 9.6 follicles), there were more follicles >6-mm diameter on the ovaries of cows with two waves on Days 3 and 4. This difference was associated with a shorter interval from wave emergence to peak concentrations of inhibin A during the first wave in two-wave cows (2.0 vs. 3.8 days; P = 0.03) and with higher peak concentrations (474 vs. 332 pg/ml; P = 0.03). Differences in peak FSH concentrations were not significant (1.7 vs. 1.3 ng/ml; P = 0.10) and were inversely related to inhibin A concentrations. The peak concentrations of inhibin A and FSH in the second nonovulatory wave in the three-wave cows were similar to the low concentrations measured in the first wave (292 vs. 332 pg/ml of inhibin A, 1.3 vs. 1.3 ng/ml of FSH; P > 0.20). Average peak concentrations of inhibin A and FSH were similar during the ovulatory wave for cows with either two or three waves in a cycle (432 vs. 464 pg/ml of inhibin A, 2.3 vs. 2.1 ng/ml of FSH; P > 0.3). The lower concentrations of FSH during the emergence of the first follicular wave in cows with three-wave cycles may have reduced the rate of development of some of the follicles and reduced the concentrations of inhibin A. This pattern of lower concentrations of FSH and inhibin A was repeated in the second nonovulatory wave but not in the ovulatory wave. Subtle differences in the concentrations of these two hormones may underlie the mechanism that influences the number of waves of ovarian follicle development that occur during the bovine estrous cycle.     

Reproductive hormone profiles in mares during the autumn transition as determined by collection of jugular blood at 6 h intervals throughout ovulatory and anovulatory cycles

The aim was to define precisely the FSH secretion pattern in mares during the two ovulatory cycles before, and for 24 days after, the last ovulation of the season and to compare this with the profiles of other reproductive hormones and follicular growth to identify changes which may lead to the termination of follicular cycles. Jugular blood was collected every 6 h from ten light horse mares for 6 weeks in autumn. Samples were assayed for FSH, LH, prolactin, inhibin, oestrone conjugates and progesterone. Luteolysis occurred earlier and periovulatory oestrone, but not inhibin, concentrations were significantly lower in the last than in the second to last cycles. In ovulatory and anovulatory cycles, daily mean FSH concentrations were low at the expected time of ovulation and high between days 9 and 11 (day 0 = ovulation), which were usually after luteolysis. However, the periovulatory FSH nadir was prolonged in the last compared with the second to last cycles, and the difference between peak and trough values was not significant in anovulatory cycles. Between day 5 and day 8, the FSH interpulse interval was approximately 2 days, and did not vary in successive cycles. The LH profile also showed progressive changes as mares entered acyclicity; the surge terminated sooner in the last than in the second to last cycles, and failed to occur when expected in acyclicity. Sporadic prolactin pulses occurred at luteolysis in a similar proportion of ovulatory and anovulatory cycles. These results indicate that inadequate gonadotrophin stimulation in early dioestrus may be a critical event leading to suboptimal follicular and luteal development, and eventually acyclicity. Moreover, the time relationships amongst changes in pituitary and ovarian hormones and follicular growth become increasingly disrupted during the autumn transition, which may contribute to the cessation of cyclicity.     

Follicular fluid adrenomedullin concentrations in spontaneous and stimulated cycles: relationship to ovarian function and endothelin-1 and nitric oxide

The objective of this study was to determine concentration of adrenomedullin (AM) in follicular fluid and whether a correlation exists between AM and nitric oxide (NO) or endothelin-1 (ET-1) levels in follicular fluid, serum 17beta-estradiol or other parameters of ovarian function in spontaneous and gonadotrophin stimulated ovarian cycles. Follicular fluid samples were obtained at oocyte retrieval from 50 women who underwent an in vitro fertilization (IVF) program: 40 undergoing ovarian hyperstimulation with recombinant FSH and 10 had spontaneous ovarian cycles. AM, ET-1, and NO were detected in all of the follicular fluid samples and their concentrations were similar in spontaneous and stimulated cycles. In patients undergoing ovarian stimulation, follicular fluid AM levels correlated with serum 17beta-estradiol concentration. No correlation was found between follicular AM concentration and parameters of ovarian function. Similarly, no relationship was observed between ET-1, NO, and AM follicular fluid concentrations in either spontaneous or stimulated cycles.This study suggests a possible regulatory effect of the sexual hormones on AM production by the ovary during the ovulatory process. The site of AM secretion and its function (if any), however, remain to be established.     

Inhibin-like activity in ovarian venous serum after unilateral ovariectomy in prepubertal gilts

To examine a role for inhibin in compensatory ovarian hypertrophy after unilateral ovariectomy (ULO) of prepubertal gilts, changes in inhibin activity in ovarian venous blood were estimated by bioassay. Three groups of 130-day-old gilts were unilaterally ovariectomized after collecting blood from an ipsilateral ovarian vein (Day O); blood samples were obtained from the remaining ovary on Day 2, 4, or 8. Coetaneous gilts underwent sham ovariectomy on Day 0, and venous blood was collected from both ovaries on Day 2, 4, or 8. An assay for inhibin activity, which measured inhibition of secretion of follicle-stimulating hormone (rFSH) by rat pituitary cells in culture, was validated for serum samples. Presumptive inhibin activity was always greater in ovarian venous serum than in peripheral serum samples. In the ULO groups, inhibin activity (in terms of a house reference preparation) in ovarian venous serum was 55 +/- 13 micrograms/ml (means +/- SE, n = 13) on Day 0, 251 +/- 79 (n = 5) on Day 2, 275 +/- 111 (n = 4) on Day 4, and 68 +/- 14 micrograms/ml (n = 4) on Day 8. The five-fold increases on Days 2 and 4 were significant (p less than 0.05). In contrast, no significant differences in inhibin activity were detected between ovarian venous serum (within gilts) or between Days 2, 4, and 8: 82 +/- 29, 73 +/- 30, and 99 +/- 48 micrograms/ml (n=4/day) in control groups. These results demonstrate that, in prepubertal gilts, the remaining ovary's response to ULO includes a major increase in release of inhibin-like activity.     

Elevated inhibin concentration in the follicular fluid of dairy cows with chronic cystic ovarian disease

This study compared serum and follicular fluid inhibin and gonadotropin profiles between chronic cystic ovarian diseased (CCOD) and normal cyclic dairy cows. Blood samples and follicular fluid were collected from CCOD cows (n=15) and cyclic cows in the follicular phase of the estrous cycle (control, n=6) and analyzed for inhibin, follicle stimulating hormone (FSH) and luteinizing hormone (LH) concentrations. There was a significant increase in inhibin and a decrease in FSH and LH concentrations in the follicular fluid of CCOD cows compared with those of cyclic cows (P < 0.05). Mean serum inhibin, FSH and LH concentrations between CCOD and cyclic cows were not differnt (P > 0.05), however, there was a tendency for serum inhibin to be higher and FSH to be lower in CCOD cows compared to cyclic animals (P < 0.1). The FSH pulse frequency also was lower in CCOD cows than in cyclic cows (P < 0.05). These data suggest that increased production of inhibin from cystic follicles of CCOD cows alters pituitary FSH secretion and subsequently reduces the concentration of FSH in follicular fluid. As a result, decreased FSH stimulation at the ovarian level could ultimately lead to the reduction in follicular LH and FSH receptor concentrations, resulting in abnormal follicular steroidogenesis in CCOD dairy cows.     

An application of Lacker's mathematical model for the prediction of ovarian response to superstimulation

Introduction. A mathematical model of ovarian follicular growth is applied to the problem of predicting ovarian response in a superstimulation protocol. Methods. Fifty-four women enrolled in an ovarian superstimulation program of therapy for the amelioration of idiopathic infertility had their ovarian cycles synchronized by taking Demulen 30 for two weeks prior to the study. Daily ultrasonographic imaging, measurements of serum estradiol and doses of hMG began on day 5 after the patients stopped taking Demulen. The diameters of individual follicles were measured and followed daily. When the largest follicle attained a diameter of 19 mm, hCG was given to induce ovulation. Individual follicle growth data were fit to a mathematical model of ovarian follicle maturation and the resulting parameters were used to classify patients into low and high ovarian response groups. Results. The parameters computed from the mathematical model fit were found to be predictive of ovarian response with a sensitivity of 71% and a specificity of 70%. The parameters were also meaningful within the context of the original mathematical model and have value for determining how doses of hMG may be adjusted during the course therapy to increase the ovarian response in individuals. Conclusion. Mathematical modeling of ultrasonographically derived follicular growth data has significant potential for clinical application in ovarian superstimulation protocols. The method of fitting follicular growth data to a mathematical follicle maturation surface furthermore provides a straightforward approach for the characterization of ovarian follicular dynamics in general.     

Antiprogestagen RU486 prevents the LH-dependent decrease in the serum concentrations of inhibin in the rat

Summary 1. In the rat, the LH-dependent ovarian progesterone rise mediates several actions of the primary surge of LH on the ovary. This experiment was aimed at elucidating the effects of the antiprogestagen RU486 on the LH-dependent decrease in both the serum concentrations and the ovarian content of inhibin.2. All rats in this experiment were treated with an antagonist of LHRH (1 mg/200 µl saline at 0800 h in proestrus) to supress the endogenous release of LH. One group of rats received 32 µg LH/250 µl saline at 1200 h in proestrus. Other group was given 4 mg RU486/200 µl oil at 0800 h in proestrus. The third group was injected with both RU486 and LH. Rats from the control group were injected with 250 µl saline and 200 µl oil. Animals were decapitated at 1700 h in proestrus and trunk blood and ovaries collected to determine the serum concentrations of LH, FSH, progesterone, 17ß-estradiol and inhibin as well as the ovarian content of inhibin.3. The ovulatory dose of LH in LHRHa-treated rats decreased both the serum concentrations and the ovarian content of inhibin and increased the serum concentrations of FSH. The administration of RU486 blocked the effect of LH on the serum concentrations of inhibin but not that on the ovarian content of inhibin.4. Since the antiprogestagen RU486 blocked the effect of LH on the serum concentrations of inhibin, we conclude that ovarian progesterone, besides mediating the effects of the primary LH surge on the ovulatory process and luteinization, participates in the LH-dependent drop in the serum concentrations of inhibin in proestrous afternoon.     

Anti-ovary antibodies after attempts at human in vitro fertilization induced by follicular puncture rather than hormonal stimulation.

Anti-ovary antibodies (AOA) have been detected in serum samples of women undergoing in vitro fertilization (IVF). High concentrations of these antibodies have been found in women who have had several IVF attempts and they appear to correlate with reduced chances of pregnancy. In this paper, AOA were assayed sequentially in a series of 140 IVF candidates to investigate the respective roles of hormonal stimulation and follicular puncture in inducing the autoimmune response. Serum was obtained 8 days after the beginning of ovarian human menopausal gonadotrophin (hMG) stimulation, then 15 days after follicular puncture. Significantly higher concentrations of IgG (P < 0.0001) AOA were observed in the second series of samples than in the first, suggesting that ovarian trauma and not hormonal stimulation is responsible for triggering antibody production. In the whole group, there was a negative correlation between IgM levels after puncture and oocyte numbers (P < 0.05). Among 'immune-responder' women, the concentrations of IgA AOA (P = 0.01) in the first sample, and of IgG (P = 0.01) or IgA AOA (P < 0.05) in the second, correlated with fewer oocytes after stimulation. There was no variation in the mean concentrations of AOA in women who achieved pregnancy.     

Elevated ovarian expression and serum concentration of α inhibin in the luteal phase during follicular development in the squirrel monkey (Saimiri boliviensis) compared to the human

The goal of the present investigation was to determine in the squirrel monkey the source and pattern of inhibin, a hormone known to effect reproductive steroid levels via pituitary and ovarian mechanisms. Since this seasonally polyestrous species is known to have elevated serum levels of reproductive steroids compared to other primates, the levels of ovarian alpha subunit mRNA expression and serum total alpha inhibin, estradiol, progesterone, and luteinizing hormone were measured and compared to human levels. Expression of the alpha subunit was robust in monkey luteal tissue compared to expression in human luteal tissue. Squirrel monkey serum inhibin peaked 4 days after the luteinizing hormone surge and correlated with progesterone changes. These luteal serum levels of inhibin were greater than 12 times higher than the human levels yet bio‐LH activities were less than in the human during the luteal phase. Inhibin concentrations during the non‐breeding season were generally half the levels measured in the breeding season and undetectable in ovariectomized animals. However, exogenous FSH stimulation induced a marked rise in inhibin, which correlated with an estradiol rise. In conclusion, abundant alpha inhibin subunit expression in the luteal ovary of the squirrel monkey and loss of serum delectability in ovariectomized animals indicates that the principle source of inhibin in the squirrel monkey is the ovary. Elevated serum inhibin levels during the luteal phase concurrent with ovulatory‐size follicular development is unique among species studied thus far. Possible simultaneous inhibin production from both follicular and luteal tissue may be responsible for the exceptionally high inhibin levels. Am. J. Primatol. 47:165–179, 1999. © 1999 Wiley‐Liss, Inc.     

Studies on GnRH agonist suppression of estrogen production in patients with endometriosis

The chronic administration of GnRH agonists to women results in the reversible suppression of estrogen production by the ovary. In the present study, the mechanism of the GnRH agonist suppression of estrogen production was investigated in patients with endometriosis. During the treatment with intranasal buserelin spray, the concentration of serum estradiol-17 beta (E2) was suppressed to near-castrate levels. Despite this marked suppression of serum E2, immunoreactive LH and FSH levels in serum were not changed. On the other hand, serum bioactive LH was markedly reduced. It was also observed during the treatment that the pituitary LH pulse disappeared and pituitary response to exogenous GnRH was significantly suppressed. In contrast, ovarian response to human menopausal gonadotropin (hMG) was not altered during the treatment. These findings suggest that the GnRH agonist suppression of estrogen production in the patients with endometriosis is through both suppression of the secretion of biologically active LH and the reduction of the LH pulse, but not through a direct inhibitory effect on ovarian estrogen biosynthesis.     

The effect of exogenous gonadotropins on ovarian function in goats actively immunized against inhibin

The aim of this investigation was to compare the ovarian response to superovulatory treatments in does before and after inhibin immunization, with a view to optimizing the superovulatory potential of the caprine ovary. To avoid interference by the ovarian cycle, the experiment was conducted out-of-season. At the onset of the experiment 48 does were subjected to treatment with an sc implant of the progestogen norgestomet, combined with a gonadotropin; eight does each received a single injection of 1200 IU eCG, 400 IU eCG or 2 mL physiological saline (control) or six injections (at 12 h intervals) constituting 16 or 5.4 AU pFSH. The does were mated and subjected to embryo collection 6 to 7 d later. Throughout the experiment ovarian function (by ultrasonography) and plasma levels of inhibin antibodies and progesterone were monitored. Of 40 does treated during the first part of the experiment, 48% showed estrus. The ovarian response in does treated with a high or low dose of eCG or a low dose of pFSH was barely in excess of the ovarian response in the saline-treated controls, whereas a superovulatory dose of pFSH (16 AU) gave a satisfactory response of, on average, 14.5 ovulations (yielding 8.8 flushed ova and embryos). Immediately after the does had been subjected to embryo collection they were actively immunized against inhibin by administering two injections of a recombinant α-subunit of ovine inhibin at four week intervals. All immunized does produced antibodies with the maximal titer reached two weeks after the second injection. Groups of immunized does were subjected to the same gonadotropin treatments as before (avoiding allocation of individuals to the same treatments). This time all does showed estrous symptoms. The ovulatory response to the various treatments, including the saline controls, was virtually identical, the overall average being 21.8 follicles and 9.1 ovulations. The average embryo yield per doe was 5.7. The results imply that inhibin acted as the key factor in determining the ovulatory response since no impact of any of the supplementary gonadotropins was noted in inhibin-immunized does. This finding gives rise to the notion that inhibin antibodies may act primarily by an intraovarian paracrine action rather than by reducing the suppressive action of inhibin on pituitary FSH release. Further, these findings confirm earlier reports that eCG is less suitable than FSH for inducing superovulation in goats, and indicate that active immunization against inhibin may be considered a viable alternative to using exogenous gonadotropin for inducing superovulation in goats.     

The effect of the presence and pattern of luteinizing hormone stimulation on ovulatory follicle development in sheep

The aim of this study was to examine the role of LH on the growth of the large preovulatory follicle and its secretion of hormones in sheep. Ewes with ovarian autotransplants were treated with GnRH-antagonist at the time of luteal regression and different LH regimes applied for 60-66 h before administration of an ovulatory stimulus (hCG). In Experiment 1 (N = 24; n = 8), ewes received either no LH or constant or pulsatile infusion of LH at the same dose (1.25 microg/h). In Experiment 2 (N = 12, n = 6), LH was constantly infused at a rate of 1.25 microg or 2.5 microg oLH/h. In Experiment 1, animals receiving either pulsatile or constant LH exhibited increases in estradiol and inhibin A secretion (P < 0.001) and a depression in FSH (P < 0.001) that resembled the normal follicular phase. Similarly in Experiment 2, doubling the dose of LH resulted in a two-fold increase in ovarian estradiol secretion (P < 0.05) but no other changes. All animals receiving LH, regardless of the pattern of stimulation, ovulated and established a normal luteal phase. In contrast, no LH treatment resulted in constant immuno-active LH without pulses, unchanged FSH and inhibin A concentrations (P < 0.05), and basal estradiol secretion (P < 0.001). Morphologically normal large antral follicles were observed in this group and although corpora lutea formed in response to hCG, progesterone profiles were abnormal. In conclusion, these results suggest that LH is an essential requirement for normal ovulatory follicle development and subsequent luteal function and show that a pulsatile mode of LH stimulation is not required by ovulatory follicles.     

Circulating concentrations of inhibin-related proteins during the ovulatory cycle of the domestic fowl (Gallus domesticus) and after induced cessation of egg laying

Circulating inhibin A, inhibin B, activin A, total immunoreactive inhibin alpha-subunit (ir-alpha inhibin), LH, FSH and progesterone concentrations were measured throughout the normal ovulatory cycle and after cessation of egg laying induced by feed restriction to investigate the potential involvement of inhibins and activins in the ovulatory cycle of the domestic hen. Plasma inhibin A varied significantly (P < 0.05) during the ovulatory cycle; the concentration was highest at the preovulatory LH surge and reached a nadir 10 h later, at about the time the F(2) follicle makes the transition to become the new F(1) follicle. Plasma FSH concentrations did not change significantly throughout the cycle and showed no correlation with inhibin A. Total ir-alpha inhibin concentrations were much higher than those of inhibin A at all stages of the ovulatory cycle and showed no correlation with inhibin A or FSH. Plasma concentrations of inhibin B and of activin A were below the detection limit of the assays in all plasma samples analysed. In the feed restriction study, plasma inhibin A and total ir-alpha inhibin showed little change until the last day of oviposition (day 0) after which they fell significantly (P < 0.05) and remained low to the end of the experiment (approximately 70-78% decrease relative to day -4). Conversely, plasma FSH increased after cessation of laying and was significantly higher (P < 0.05) from day 3 to the end of the study (approximately 50% increase on day 6 relative to day -4). Plasma FSH values were negatively correlated with inhibin A (r = -0.39; P < 0.005) and total ir-alpha inhibin (r = -0.36; P < 0.005). Plasma LH and progesterone also decreased (P < 0.05) during feed restriction. The decrease in LH preceded the terminal oviposition and the associated fall in inhibin A by 2 days; there was a positive correlation between LH and inhibin A (r = 0.35; P < 0.005). Taken together these findings support (i) a role for LH in promoting inhibin A secretion by preovulatory follicles and (ii) an endocrine role for inhibin A secreted by preovulatory follicles in the maintenance of tonic FSH secretion in laying hens.     

Effect of inhibin immunisation using different synthetic peptide fragments of the bovine αc-subunit on plasma anti-inhibin titres,plasma FSH concentrations and the incidence of multiple ovulation in heifers

In an effort to improve the effectiveness of inhibin immunisation in promoting multiple ovulation in cattle and to clarify the mechanism(s) involved, heifers (n = 5 per group) were immunised against ovalbumin conjugates of different synthetic peptide sequences of the α_c-subunit of bovine inhibin (bIα) selected using antigenic prediction methods. Plasma inhibin antibody titre (percentage binding of ¹²⁵I-labelled M_r 32 000 native bovine inhibin), plasma follicle-stimulating hormone (FSH) concentration and ovulatory response (number of corpora lutea observed by transrectal ovarian ultrasonography) were recorded over a 16 week period.Heifers immunised against the bIα1–29 and 63–72 peptides (alone or in combination), had relatively high anti-inhibin titres (over 7.5% binding) and showed a significantly (P < 0.05) increased incidence of multiple ovulations (18–65%) compared with ovalbumin-immunised controls. However, immunisation against the 1–16 and 108–123 peptides was relatively ineffective in generating antibodies reactive with native inhibin (less than 7.5% binding) and gave little or no increase in incidence of multiple ovulations (0–10%).Analysis of results for all 33 heifers revealed a significant linear relationship between mean inhibin antibody titre and mean plasma FSH concentration (r = 0.42; P < 0.02) and between mean inhibin antibody titre and incidence of multiple ovulation (r = 0.89; P < 0.0001). A significant quadratic relationship existed between mean inhibin antibody titre and the mean number of ovulations per cycle (r = 0.88; P < 0.0001). However, partial correlation analysis showed a highly significant association between anti-inhibin titre and ovulatory response which was independent of changes in mean plasma FSH concentrations.These results extend previous studies involving inhibin peptide-immunised cattle by showing that the magnitude of the ovulatory response is directly related to the prevailing titre of antibodies reactive with native inhibin. However, they do not support the hypothesis that the ovulatory response is mediated solely by a rise in FSH secretion.     

Continuous subcutaneous infusion of human menopausal gonadotropin in anovulatory women with decreased FSH/LH ratio and androgen excess

Ten women of polycystic ovarian-type (PCO-type) anovulation, having a decreased ratio of FSH to LH and androgen excess, resistant to the previous clomiphene, bromocriptine and/or daily im injections of human menopausal gonadotropine (hMG), were treated with continuous sc infusion of 150 IU/day hMG. The treatment was initiated on cycle day 2-5 and continued until the dominant follicle reached 20 mm or more in diameter, when an im bolus of 10,000 IU human chorionic gonadotropin was given. The treatment elevated the geometric mean of pretreatment serum FSH (8.6 mIU/ml) to 15.9 mIU/ml (p less than 0.001), while serum LH decreased from 29.4 mIU/ml to 20.7 mIU/ml (p less than 0.01). This resulted in a highly significant increase in the FSH/LH ratio from 0.29 to 0.77 (p less than 0.0001). Follicle enlargement was demonstrated in 13 of the 14 treatment cycles, 12 of which were ovulatory. Pregnancy ensued in 4 cases, 1 of which was a quadruplet pregnancy. Continuous infusion of hMG was indicated as an effective way of inducing ovulation in PCO-type anovulation resistant to conventional methods of ovulation induction.     

Prolactin concentrations in follicular fluid following ovarian hyperstimulation for in vitro fertilization

Prolactin (PRL) and sex steroid concentrations were measured in follicular fluids of women treated either with (1) clomiphene/hCG or with (2) clomiphene + hMG/hCG. Method 1 of ovarian stimulation resulted in lower follicular PRL and higher oestradiol-17 beta (E2) and progesterone (P) concentrations than method 2. There was no difference in the PRL and sex steroid concentrations of follicles with fertilized and of those yielding unfertilized ova, but in both stimulation types, follicles from which no oocytes were obtained had high PRL and low E2 and P levels. Significant positive correlations were evident for PRL and T and E2 and P, respectively, while PRL and P were negatively correlated.     

Is changing hypothalamic activity important for control of ovulation?

The activity of the hypothalamic gonadotrophin releasing hormone pulse generator in women with regular ovulatory and anovulatory menstrual cycles was assessed to see whether changes therein are important determinants of normal and impaired ovarian function. Endogenous gonadotrophin releasing hormone secretion was inferred by measurement of the pituitary luteinising hormone response by characterisation of pulsatile luteinising hormone release over eight hours on three occasions during the course of follicular development and once during the luteal stage of the same cycles. In 13 ovulatory cycles (serum progesterone concentration greater than 25 nmol/l) confirmed by ovarian ultrasonography a pronounced variability in luteinising hormone pulse patterns among subjects was compatible with ovulation. In the luteal stage of ovulatory cycles the luteinising hormone interpeak interval (85 min, range 42-125) was significantly longer than that during the early follicular (64 min, 40-103), mid-follicular (62 min, 37-107), and late follicular (59 min, 39-80) stages of the same cycles. Thus in ovulatory cycles no increase in frequency of the gonadotrophin releasing hormone pulse generator was detected during follicular development, though this activity decreased in the luteal stage. In five late follicular stage studies in which part of the preovulatory luteinising hormone surge was captured no change in pulse frequency of luteinising hormone was detected compared with the mid-follicular stage of the same cycles or when compared with the late follicular stage of other cycles when no luteinising hormone surge was captured. Though mean luteinising hormone concentrations in luteinising hormone surge series (36 IU/l) were high, the amplitude of luteinising hormone pulses (165%) was only slightly greater than during non-surge late follicular stage studies (145%). Hence no change in hypothalamic gonadotrophin releasing hormone activity is required to generate the preovulatory discharge of luteinising hormone in man, which occurs as a result of the sensitising action of rising oestradiol concentrations on pituitary responsiveness to the same hypothalamic input signal. Luteinising hormone pulse frequency, peak amplitude, and mean serum luteinising hormone concentrations in seven anovulatory cycles (progesterone concentration less than 10 nmol/l) were not different from those at comparable stages of ovulatory cycles. These data suggest that the primary abnormality in this group of regularly menstruating anovulatory women lies in the ovary rather than in the hypothalamic control of the anterior pituitary.     

Serum concentrations of deoxycorticosterone in women during the luteal phase of the ovarian cycle are not suppressed by dexamethasone treatment

We determined the serum levels of deoxycorticosterone (DOC) in plasma of six healthy, apparently ovulatory women during the mid-follicular and mid-luteal phases of their ovarian cycles; and we evaluated the effect of dexamethasone (1 mg by mouth) on the concentrations of DOC and cortisol in serum at times when plasma progesterone levels were high or low. The serum levels of DOC, unlike those of cortisol, did not vary significantly in single blood samples obtained in the morning (8-10 a.m.) and afternoon (3-5 p.m.); and serum DOC levels in women were significantly higher (P less than 0.05) during the mid-luteal phase than during the mid-follicular phase of the cycle. There were unmistakable diurnal variations in serum levels of cortisol, and cortisol concentrations were reduced to less than 20% of pretreatment levels after the ingestion of 1 mg dexamethasone during the mid-follicular or mid-luteal phase. The serum concentrations of DOC were reduced only to approx 70% of pretreatment levels after dexamethasone ingestion during the follicular phase. The serum levels of DOC did not decline significantly after administration of dexamethasone during the mid-luteal phase, when progesterone levels in serum are high (14-16 ng/ml). Blood samples also were obtained at hourly intervals during the 24 h before and after dexamethasone administration in one woman during the follicular phase and in another woman the during the early luteal phase (progesterone levels = 1-3 ng/ml) of the ovarian cycle. DOC levels (pre-dexamethasone) fluctuated in synchrony with those of cortisol in the woman studied during the follicular phase but not in the woman studied during the early luteal phase of the cycle. In the post-dexamethasone period, plasma cortisol levels were suppressed for at least 24 h in both women whereas DOC levels were decreased only partially. We conclude that plasma DOC is derived from both adrenal secretion and from extraadrenal 21-hydroxylation of progesterone--the latter source of DOC is not affected by dexamethasone suppression of ACTH secretion.     

Effect of peptide to carrier ratio on the immune and ovarian response to inhibin immunization in cattle

We report on the effects of the peptide to carrier ratio on the immune and biological response to inhibin immunization in cattle. A peptide sequence from the α_C-subunit of bovine inhibin was synthesized and conjugated to human serum albumin (HSA) at ratios of 4.3 moles (L) and 13.1 moles (M) of peptide per mole of HSA. Hereford-cross heifers (n = 6 per group) were injected with 3 mg of one of the peptide conjugates at primary, followed by a booster injection (1.5 mg) 11 weeks later. Control heifers (n = 6) were injected with HSA only. Blood samples were taken at regular intervals to measure antibody titre. Ovulation rate was measured by ultrasonography. Antibodies were generated in both peptide immunized groups. Control heifers and group L heifers had 1 ovulation at all ovulatory cycles monitored. Ovulation rate was increased (P < 0.05) in group M immunized heifers, with four of six heifers having twin ovulations in the first cycle following boost. These data support those of previous studies which indicated that immunization against the α_C-subunit of bovine inhibin significantly disrupted the mechanism(s) controlling ovulation rate in cattle. It also indicates that both the immune and associated biological response is dependent on the nature of the conjugate used for immunization, specifically the ratio of peptide to carrier.     

Successful pregnancy in a woman with ovarian failure associated with mutation in the beta-subunit of luteinizing hormone.

BACKGROUND: We report a successful pregnancy in a woman with severe ovarian dysfunction and infertility associated with a variant beta-subunit of luteinizing hormone (LH). METHOD/OUTCOME: A 35-year-old woman consulted our unit for infertility. Laparoscopy and ultrasonography showed obstruction of the right tube and ovulation from the right ovary only. Human menopausal gonadotrophin (hMG) therapy was used for six subsequent cycles, but did not result in conception. Subsequently, marked elevation of follicle-stimulating hormone (FSH) and testosterone, together with polycystic ovary (PCO) were noted. The patient failed to respond to ovarian stimulation by hMG. Severe ovarian dysfunction such as premature ovarian failure (POF) was strongly suspected. Sequence analysis of the LH beta-subunit gene indicated heterozygosity for point mutations Trp(8) to Arg(8) and Ile(15) to Thr(15) in the coding sequence. LH hypersecretion resembling that seen in PCO syndrome was observed. Induction of ovulation by hMG was successful in the first cycle in which the basal LH and FSH were well controlled with gonadotrophin-releasing hormone analog following estrogen-progesterone replacement. She conceived and delivered a healthy male infant at term. CONCLUSION: Clinicians should be clinically aware of patients with immunologically anomalous LH variant who might be at risk of developing ovarian failure within a relatively short time span. Pertinent treatment should be applied without delay in such cases.