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Effect of exogenous hormones on the ovulation of primary and secondary oocytes in LT/Sv strain mice.

LT/Sv strain mice ovulate both primary and secondary oocytes. These are fertilizable and give rise to digynic triploid and normal diploid conceptuses, respectively. A previous study [Kaufman and Speirs, 1987] had indicated that just over 20% of embryos recovered on the 10th day of gestation from spontaneously ovulating females had a triploid chromosome constitution. This value was considerably lower than might have been expected by extrapolation from earlier studies in which LT/Sv mice had been given exogenous gonadotrophins. In the present study, therefore, cytogenetic analysis of fertilized eggs was performed at the first cleavage mitosis in (1) spontaneously ovulating females mated to F1 hybrid males, and (2) superovulated females mated to similar males. Additional females from group (1) were autopsied on the 10th day of gestation, and the ploidy of embryos isolated at this stage of gestation was determined. Exposure to exogenous gonadotrophins significantly increased the proportion of eggs that were ovulated as primary oocytes (34.4%), compared to the situation observed following spontaneous ovulation (24.4%). All the triploids encountered in both series were of the digynic type and characteristically (for LT/Sv mice) had an oocyte-derived set with 40 chromosomes present, and a sperm-derived set containing 20 chromosomes. Similar numbers of eggs were recovered from spontaneously ovulating females on the 1st and 10th days of gestation, and the incidence of triploidy observed on the 10th day was 22.1%. The influence of exogenous hormones in increasing the "spontaneous" level of triploidy in LT/Sv and in other strains of mice is briefly reviewed.

Animals↗

The POU gene ceh-18 promotes gonadal sheath cell differentiation and function required for meiotic maturation and ovulation in Caenorhabditis elegans.

In Caenorhabditis elegans, specialized contractile myoepithelial cells of the somatic gonad, the gonadal sheath cells, are closely apposed to oocytes and are required for normal meiotic maturation and ovulation. Previously we found that mutations in the ceh-18 gene, which encodes a POU-class homeoprotein expressed in sheath cells, result in oocyte defects. To determine the basis for these oocyte defects, we have used time-lapse video Nomarski microscopy to observe meiotic maturation, ovulation, and early embryogenesis in ceh-18 mutants. In ceh-18 mutants sheath cell contractions are weaker, less frequent, and uncoordinated throughout the sequence of ovulation events, and ovulation is defective. Defective ovulation can result in the formation of endomitotic oocytes in the gonad, the formation of haploid embryos, and reversals in embryonic polarity. ceh-18 mutant oocytes exhibit defects prior to nuclear envelope breakdown, suggesting that they are physiologically different from the wild type. We observed delays in meiotic maturation, as well as maturation out of the normal spatial and temporal sequence, suggesting that proximal sheath cells directly or indirectly promote and spatially restrict meiotic maturation. Analysis of sheath cell differentiation in ceh-18 mutants using antibodies to proteins of the contractile apparatus reveals that although contractile proteins are expressed, the sheath cells appear disorganized. Transmission electron microscopy reveals that ceh-18 mutant sheath cells are morphologically irregular and only loosely cover oocytes. Taken together, these observations indicate that ceh-18 is a crucial determinant of sheath cell differentiation, a function required for normal meiotic maturation and ovulation.

Animals↗

Increase in ovarian alpha-inhibin gene expression and plasma immunoreactive inhibin level is correlated with a decrease in ovulation rate in the domestic hen.

This study was designed to examine the relationship among ovulation rate, inhibin alpha-subunit gene expression, and plasma immunoreactive inhibin levels between hens laying at a high ovulation rate and hens laying at a low ovulation rate. A porcine inhibin alpha-subunit cDNA was used to examine the expression of inhibin alpha-subunit in the granulosa cell layers of preovulatory chicken follicles. Total RNA was extracted from granulosa layers of hens laying at a high and low rate and used in slot blot analyses. A dose-related increase in signal intensity with increasing doses of RNA for hens laying at a high and low rate was observed. The expression of the alpha-subunit of inhibin (per microgram of RNA) in granulosa layers of hens laying at a low rate was found to be greater than that in hens laying at a high rate (P < 0.001). Different groups of hens laying at the same rates as those used in the RNA determinations were selected for determination of plasma inhibin levels. Plasma samples from those hens laying at a high and low rate were collected from 1000 to 1800 hr at 2-hr intervals. Plasma inhibin, luteinizing hormone (LH), and follicle stimulating hormone (FSH) were determined by RIA. Plasma inhibin level was higher in hens laying at a low rate than that in those laying at a high rate (P < 0.05), while plasma LH level in hens laying at a low rate was lower than that in hens with a high ovulation rate (P < 0.01). There was no difference in plasma FSH level associated with rate of lay. Results in this study suggest that the ovulation rate-related changes in inhibin alpha-subunit gene expression and plasma immunoreactive inhibin level do not directly affect ovulation rate through a modulation of plasma FSH level.

Animals↗

Ovarian epithelial dysplasia in relation to ovulation induction and nulliparity.

OBJECTIVE: The goal of this study was to assess the relationship between ovulation induction, nulliparity, and ovarian epithelial dysplasia. METHODS: This retrospective cohort study was performed in one teaching and one district general hospital in London. The subjects, 83 women who had undergone hysterectomy and bilateral oophorectomy and whose ovaries were reported as "normal," were divided into three groups: ovulation induction (13), nulliparity (20), and fertile controls (50). These ovaries were independently reviewed by two pathologists who assigned a score of 0, 1, or 2 to nine epithelial cytological and architectural features. The main outcome measure was the total dysplasia score, which was used to quantify the degree of ovarian epithelial abnormality in the three groups. RESULTS: The mean dysplasia score was significantly higher in the women who had undergone ovulation induction than in the fertile controls (7.92 vs 5.70, P = 0.012). The magnitude of the difference between the ovulation induction group and controls remained similar after adjusting for age, parity, and duration of oral contraceptive use (2.17, 95% CI: -0.11-4.44). However, the statistical significance of this difference was reduced (P = 0.062). We did not find any evidence of a difference in dysplasia score between nulliparous women and controls, neither before (P = 0.85) nor after adjusting for age and duration of oral contraceptive use (P = 0.87). CONCLUSIONS: These results suggest a possible association between ovarian epithelial dysplasia and ovulation induction therapy, in accord with previous reports of increased risk of ovarian cancer in women with a history of fertility treatment. The higher dysplasia score could be attributable to the drugs used to induce ovulation or to a genetic susceptibility to ovarian cancer.

Adult↗

Influence of the formamidine pesticide chlordimeform on ovulation in the female hamster: dissociable shifts in the luteinizing hormone surge and oocyte release.

The formamidine pesticide chlordimeform (CDF) has been found to interfere with the hormonal control of ovulation in the rat by a presumptive disruption in the catecholaminergic regulation of the midcycle surge of luteinizing hormone (LH). While the brain hypothalamic mechanisms underlying generation of the hamster surge have not been as well-defined, there is evidence for an adrenergic component. The present experiments were designed to investigate the effects of CDF on ovulation in the golden hamster and whether any observed alterations are associated with differences in the appearance of the surge. Intraperitoneal CDF injections (0, 75, 150, or 200 mg/kg) at 1400 hr on proestrus caused a dose-related reduction in oocytes retrieved at 0700 hr on the day of estrus. Additional experiments demonstrated that this effect was due to a delay in ovulation and not a decrease in the complement of oocytes released. Doses of 150 or 200 mg/kg at 1100 hr or 200 mg/kg at 1600 hr were without a comparable ovulatory effect. Characterization of the LH surge indicated that there was a dose-related delay, but that this effect was not sufficient to account for the delay in ovulation. Animals dosed at 1100 hr (150 or 200 mg/kg) exhibited shifts in the surge identical to those animals injected at 1400 hr, but did not show any such detectable effects on the time of oocyte release. CDF administered at 1100 or 1400 hr also caused alterations in serum estradiol and progesterone, even at a relatively low dose of 37.5 mg/kg. The results indicate that CDF given at a time just prior to the preovulatory rise in LH is able to impede ovulation in the hamster by possibly influencing some ovulatory event(s) triggered by the surge. Also, the ability of CDF to alter the timing of LH release is consistent with catecholaminergic participation in the generation of the gonadotropin signal stimulating the final events leading to ovulation. Under the present conditions, this perturbation can be characterized as a delay in the LH trigger, rather than a blockade.

Animals↗

Endocrine disruption by indole-3-carbinol and tamoxifen: blockage of ovulation.

Immature Sprague-Dawley rats received daily doses of indole-3-carbinol (I3C, 0-1.5 g/kg/day), 3,3'-diindolymethane (DIM, 0-400 mg/kg/day), tamoxifen (TAM, 0-0.5 mg/kg/day), or vehicle to determine if their antiestrogenic effects occur by the same mechanism and whether I3C's action is mediated by DIM. Follicular development was induced on day 24 of age by equine chorionic gonadotropin (eCG, 5 IU) 1 day after the initial dose. In a hormone replacement study, human chorionic gonadotropin (hCG, 10 IU sc, 48 h post-eCG) was used to mimic a normal preovulatoy luteinizing hormone (LH) surge following treatment with either I3C or TAM. Blood and ovaries were collected throughout follicular development and the number of ova shed was measured on the morning following expected ovulation (72 h post-eCG). I3C but not TAM reduced body weight gain at higher doses after 4 days of dosing. Ovarian weight gain and ovulation were inhibited by both I3C and TAM in a dose-dependent fashion. During the preovulatory period, both I3C and TAM blocked normal LH and follicle-stimulating hormone (FSH) surges and suppressed serum progesterone (P(4)) profoundly without changing circulating levels of estrogen (E(2)). At the time of expected ovulation, serum E(2) was increased in rats receiving I3C or tamoxifen, whereas serum P(4) was dose-dependently decreased. DIM exerted no significant effects on any of the endpoints studied, even at the highest dose, indicating that the antiestrogenic effects of I3C are not mediated by this metabolite of I3C. hCG successfully restored ovarian weight gain and ovulation in TAM-treated rats. However, hCG only partially reversed the blockage of ovulation by I3C, although ovarian weight gain was restored to normal. In summary, both I3C and TAM block ovulation by altering preovulatory concentrations of LH and FSH, but I3C appears to exert its effect(s) by (a) different mechanism(s) of action. I3C seems to act at both the ovarian and hypothalamic levels by mechanisms similar to those seen in TCDD-treated rats, whereas TAM appears to act only on the hypothalamic-pituitary axis as an anti-estrogen.

Animals↗

Ovulation as a tissue remodelling process. Proteolysis and cumulus expansion.

Ovulation, recurring every midcycle of the mammalian female and triggered by a surge of luteinizing hormone (LH) released from the pituitary, is an essential prerequisite for fertilization and subsequent embryonic development. Here we shall describe two of the biological components of the ovulatory response, cumulus expansion (frequently denoted as cumulus maturation) and the rupture of follicular wall, both crucial for the release of a fertilizable ovum. The role of a proteolytic cascade and its regulation by eicosanoids will be emphasized in relation to follicle rupture. The new data implicating cumulus maturation as an essential step for the release of the ovum and the apparent mediatory role of interleukin-1 in this process will be presented. LH/hCG stimulates, in the preovulatory follicles, a cascade of proteolytic enzymes, including plasminogen activator (PA), plasmin and matrix metalloproteinase 1 (MMP-1). These enzymes bring about the degradation of perifollicular matrix and, most notably, the decomposition of the meshwork of collagen fibers which provides the strength to follicular wall. Furthermore, pharmacological blockage of any of these enzymes resulted in inhibition of follicle rupture. LH/hCG stimulates, in addition, an increase in ovarian production of eicosanoids. These include prostaglandins, obtained from arachidonic acid via the cyclooxygenase pathway and leukotrienes, the products of lipoxygenase. Previous studies from our and other laboratories have demonstrated the ability of inhibitors of cyclooxygenase and of lipoxygenases to suppress ovulation in several mammalian species. MK-886, which inhibits the translocation of 5-lipoxygenase (5-LO) from the cytosol and its binding to the membranal 5-LO activating enzyme, suppressed dose-dependently follicular rupture from the treated ovary. Zymographic analysis of ovarian extracts from PMSG/hCG-stimulated rats revealed a band of collagenolytic activity at 52kD, corresponding to human MMP-1 and at 72kD, corresponding to human MMP-2. Both activities were markedly stimulated by administration of hCG and were significantly inhibited by indomethacin, NDGA or MK-886. Thus, eicosanoids seem to mediate LH stimulation of follicular collagenase. Interleukin-1 (IL-1) has been recently implicated in ovulation. The ability of an IL-1 receptor antagonist (ra) to block ovulation in vivo and in vitro has been demonstrated recently. Morphological examination of the ovulatory follicles failing to ovulate suggests that this effect is exerted by inhibiting cumulus oophorus expansion and detachment from mural granulosa cells. In vitro, IL-1ra attenuated the action of hCG and FSH on cumulus expansion and follicular hyaluronic acid synthesis. Thus, IL-1 seems to mediate and/or facilitate gonadotropin action on cumulus expansion, and hence on ovulation.

Animals↗

Effect of ovarian endometriosis on ovulation in rabbits.

To study the relationship between endometriosis and ovulatory dysfunction, we induced ovarian endometriosis in the rabbit model Adipose tissue was placed in the contralateral ovary as a control. Ovulation was induced with human chorionic gonadotropin, and ovulation points were counted before and after induction of endometriosis. Periovarian adhesions were graded, and ovaries were histologically examined. A significant decrease in the number of ovulation points was observed in ovaries with endometrial tissue (p = 0.001) but not in ovaries that contained adipose tissue (p = 0.095). Periovarian adhesions decreased the number of ovulation points (p less than 0.01) in ovaries that contained adipose or endometrial tissues. Multivariate analysis demonstrated that an increase in adhesion severity was correlated with a decrease in the number of ovulation points (p less than 0.05), but endometrial tissue was not (p = 0.45). We conclude that, in the rabbit model, minimal ovarian endometriosis impairs ovulation primarily through a mechanism related to periovarian adhesions.

Analysis of Variance↗

Electrochemically stimulated release of luteinizing hormone and ovulation after surgical interruption of lateral hypothalamic connections in the rat.

Plasma LH and FSH were studied in adult female rats following bilateral electrochemical stimulation (ECS) of the medial preoptic area (MPOA). By stereotaxic surgery frontal (FC) and frontal-lateral (LFC) retrochiasmatic "deafferenting" cuts were made with a Halász knife (1.5 mm radius). At 3 and 10 weeks after surgery rats were given pentobarbital (32 mg/kg, i.p.) at 13:30h and stimulated at 15:00h with anodal direct current (20 muA for 60 sec) via concentric bipolar steel electrodes placed bilaterally 0.9 mm from the midline. Stimulation at 3 weeks after FC increased plasma LH from a prestimulation level of 95 +/- ng/ml to 227 +/- 51 ng/ml 80 min after ECS, from which it fell to 111 +/- 29 ng/ml at 160 min, with 2 of 7 animals ovulating the next day. LFC females had similar pre-ECS plasma LH levels 3 weeks after surgery (71 +/- 10 ng/ml), but LH concentrations at 80 min (104 +/- 21 ng/ml) and 160 min post-ECS were significantly lower than those of FC rats, and 0 of 5 rats ovulated. Following a similar protocol 10 weeks after surgery, stimulating the MPOA resulted in comparable elevations in plasma LH and 4 of 10 FC animals ovulated; however, the LFC group still retained a significant blocking effect on ovulation (only 1 of 12 ovulated) when compared with controls the next day. Plasma FSH concentrations were not significantly altered by MPOA stimulation at the parameters employed, either before or after deafferentation. However, LFC resulted in reduced ovarian and uterine weights when compared with controls at both 3 and 10 weeks, whereas FC exerted no observable effect on these organs at these intervals of study. The results of these studies suggest that lateral input to the media basal hypothalamus contributes to MPOA mediated release of LH and ovulation as well as to tonic maintenance of ovarian and uterine function.

Animals↗

Mechanism of participation of medial preoptic area in the hippocampal inhibition of ovulation.

Bilateral hippocampal (HPC) stimulation with anodal direct current on the afternoon of proestrus blocked spontaneous ovulation in 87.5% of the Wistar rats subjected to the experiments. The incidence of the ovulation block by this procedure was reduced to 16.7% on bilateral injection of 0.25 microgram picrotoxin into the medial preoptic area (mPOA) preceding as well as following the stimulation. Ovulation was also blocked in 53.8% of the animals by bilateral injection of 50.0 microgram GABA into mPOA, while only 20.0% animals showed a blockade of ovulation by saline injection into mPOA. These observations indicate that blockade of ovulation by HPC stimulation can be simulated by local injection of GABA into mPOA while the effect of stimulation can be blocked by local injection of picrotoxin. Thus, indicating the possibility of GABA being neurotransmitter involved at the level of mPOA for mediating the inhibitory hippocampal influence on ovulation.

Animals↗

A study of the ovulation-inhibiting effects of dexamethasone in the domestic hen.

Laying hens were injected with a series of graded doses of dexamethasone following the terminal oviposition of a sequence to determine the effect of this compound on the time of ovulation. It was found that injection of 10-500 micrograms of dexamethasone/kg inhibited ovulation but there was no evidence of an ovulation-inducing effect when the dose was reduced below 10 micrograms/kg. The results for dexamethasone are in contrast to the effects of corticosterone which inhibits ovulation at high doses and induces ovulation at lower doses providing the hens are injected at the stage of the ovulatory cycle used in this study. It is suggested that the ovulation-inducing action of corticosterone involves a target site remote from the hypothalamic-pituitary axis.

Animals↗

Effects of [D-Ala6, Pro9-NEt]-LHRH and catecholaminergic drugs on gonadotropin secretion and ovulation in the Chinese loach (Paramisgurnus dabryanus).

The effects of [D-Ala6,Pro9-NEt]-LHRH (LHRH-A) alone and in combination with drugs which influence the actions of dopamine or the synthesis of catecholamines on gonadotropin (GtH) secretion and ovulation in the loach (Paramisgurnus dabryanus) were investigated. LHRH-A alone stimulated an increase in serum GtH levels in the loach, but was a relatively ineffective treatment for the induction of ovulation. Injection of the dopamine receptor antagonist pimozide caused a marked potentiation of the GtH-release response to LHRH-A, and combined injections of pimozide and LHRH-A were an effective treatment for the induction of ovulation. Reserpine, a drug which causes depletion of catecholamines from presynaptic terminals, also caused a marked potentiation of the GtH-release response to LHRH-A and combined treatment induced ovulation. Similarly, administration of alpha-methyl-para-tyrosine to block conversion of tyrosine to L-dopa, or carbidopa to block conversion of L-dopa to dopamine, potentiated the GtH-release response to LHRH-A and induced ovulation. In contrast, the use of diethyldithiocarbamate, to block conversion of dopamine to norepinephrine, failed to augment the action of LHRH-A on GtH release and ovulation. The present results provide further evidence to suggest that dopamine functions as a gonadotropin release-inhibitory factor in teleosts, and demonstrate that the use of drugs which block either the synthesis or the actions of dopamine potentiates the action of LHRH-A in teleosts.

Animals↗

Induction of ovulation and oviposition in female quail with luteinizing hormone, luteinizing hormone releasing hormone, or progesterone.

Regularly laying female Japanese quail were injected 12 or 18 hr before the next expected oviposition with 25 micrograms oLH, 25 micrograms luteinizing hormone releasing hormone (LHRH), or 0.05 mg progesterone, and the subsequent oviposition was recorded and ovulation determined by autopsy 9 hr after the injection. Plasma progesterone levels were measured in blood collected from a wing vein during the postinjection interval. Vehicle-injected birds served as control. All treatments resulted in premature oviposition and ovulations in 50-100% of birds when injected 12 hr before the next expected oviposition. None of the vehicle-injected birds showed any advancement in either oviposition or ovulation. Premature oviposition was generally followed by premature ovulation within 1 hr, when birds were treated 12 hr before the next expected oviposition and was preceded by a rise in plasma progesterone levels which reached values similar to those occurring during the normal preovulatory period. When birds were injected 18 hr before the next expected oviposition, the incidence of premature oviposition was very low, premature ovulation was absent, and the rise in plasma progesterone levels following treatment was substantially less than in the former group. The results suggest that oviposition and ovulation in quail may be initiated by LHRH induced LH release from the pituitary gland and that progesterone may stimulate LHRH and LH release. The timing of the ovulatory cycle may depend upon the phase of follicular maturation.

Animals↗

Fertility of dairy cattle following oestrus and ovulation controlled with cloprostenol, oestradiol benzoate and progesterone or progesterone and cloprostenol.

There have been several approaches to the control of the timing of the oestrous cycle and ovulation in dairy cattle in the last three decades. The first phase involved the use of progestins which were administered in various forms for prolonged periods. Although the timing of oestrus was controlled in most animals after withdrawal of the treatment, this control was not very precise and pregnancy rates from insemination at the first oestrus after treatment were reported to be below normal. Attempts were then made to combine short-term progestin treatments with oestrogens as luteolytic agents to gain better control of the timing of oestrus and ovulation. These studies resulted in some cases in better synchronization of oestrus and improved pregnancy rates. The discovery that prostaglandin F2 alpha (PGF2 alpha) and its synthetic analogue, cloprostenol were potent luetolytic agents in the cow led in the past decade to the use of these agents for oestrus and ovulation control in cattle. Prostaglandins for this purpose are ineffective in anovulatory cows, in cows with deficient luteal function and in the first 5 days of the oestrus cycle when a new corpus luteum is being formed. This limitation in their use has encouraged investigations into the combined use of short-term progestin treatment with prostaglandins to give more effective control of the timing of oestrus and ovulation and to avoid the adverse effects on fertility of long-term progestin treatment. Short-term progestin treatment combined with prostaglandins should mean that fewer cows would have ovulation suppressed for long periods and fertility of treated cows should be improved. A comparison of three procedures of ovulation control and fertility results shows the short-term progestin treatment combined with prostaglandin to be the most effective.

Animals↗

The induction of oestrus and ovulation in seasonally anovular ewes by exposure to rams.

In anoestrus ewes of many breeds, the introduction of rams will evoke an increase in the tonic secretion of LH, pre-ovulatory surges of LH and FSH, and ovulation, provided the ewes have been preconditioned by a period of complete isolation from the rams. The "ram effect" is apparently mediated by a pheromone which seems to inhibit or reverse the effects of photoperiod on the secretion of LH pulses. In the absence of progesterone "priming" the ewes rarely show oestrus behaviour around the time of the first ovulation. In some of the ewes, the corpus luteum formed at the first ovulation is apparently normal, and the second ovulation and first oestrus is observed about one oestrus cycle later. In the remainder of the ewes, the first corpus luteum regresses prematurely, a second ovulation is seen about 6 days after the first, and the third ovulation and first oestrus are seen after a further 17 days. Progesterone treatment prior to the introduction of rams prevents the occurrence of short cycles, apparently by delaying the LH surge. The combination of the ram effect and progesterone treatments can provide useful synchrony of oestrus for artificial insemination, and also some gains in fecundity. There is considerable potential for using the effect in intensive breeding programmes.

Animals↗

Pure FSH alone induces ovulation and subsequent pregnancy in the mouse resulting in fetal development.

The role of FSH in inducing folliculogenesis is well established. Recently, the availability of pure FSH has led to a reevaluation of its role in the process of ovulation. Previously, these functions have been examined separately, usually with pregnant mares' serum gonadotropin (PMSG) followed by FSH for ovulation or FSH for folliculogenesis followed by hCG for ovulation. To determine if FSH alone can induce both folliculogenesis, ovulation and establish a functioning corpus luteum without exogenous LH, we injected sexually mature intact mice (CD-1) with either ovine FSH (oFSH, 5 micrograms; < 0.2% LH contamination) or recombinant FSH (RCFSH, 1 IU; devoid of any LH activity) to stimulate folliculogenesis, followed forty-eight hours later by a second injection of the same preparation (oFSH, 15 micrograms; RCFSH, 1 IU, respectively) to induce ovulation. Injected female mice were mated individually with a fertile male. On days 15-17, pregnancy rates and fetal development were obtained for each animal and were compared with controls, mice injected with PMSG (1 IU) followed by hCG (1 IU). oFSH/oFSH and RCFSH/RCFSH results were combined since no statistical significant differences were detected between these groups. The pregnancy rate for the group receiving FSH/FSH (78.3%, n = 23) was higher than that of the PMSG/hCG group (48.3%, n = 27; p = .02). The number of fetuses produced per mouse in animals receiving FSH alone (8.5 +/- 1.1; mean +/- S.E.) also was greater than the controls (4.5 +/- 99; p = .01). We conclude that the ability of these animals to proceed beyond ovulation to implantation with fetal development demonstrates FSH's ability to cause not only follicular maturation and rupture, but also granulosa cell luteinization, further identifying the potentially important role of FSH in the ovulatory process.

Animals↗

Re-examining steroid hormone metabolites as ovulation markers using monoclonal antibodies.

Plasma progesterone and plasma estradiol levels are commonly used to monitor ovulation in women although for the adequate documentation of ovulation the expense and discomfort of multiple venipuncture sampling may be required. Accurate and definitive information on ovulation in women can be obtained by the simple measurement of metabolites of progesterone and estradiol in early morning urine samples. These analyses have been made possible by the use of our own highly specific monoclonal antibodies to both pregnanediol-3-glucuronide (P-3-G) and estrone-3-glucuronide (E-3-G) and the development of simple, direct, automated enzyme-linked immunosorbent assays. Sequential sampling and the generation of ovulation profiles allows detection of ovulation and identification of the infertile/fertile phases of the cycle for either planned pregnancies or natural family planning. Aberrations in ovulation are easily detected as is documentation of the transition to menopause. The use of sequential, spaced, early morning urine samples for P-3-G and E-3-G allows accurate assessment of ovulatory function rather than relying on the usual single plasma sampling. The data presented also show that the direct determination of plasma pregnanediol-3-glucuronide in plasma is as informative as plasma progesterone measurement.

Adult↗

Effects of manipulating catecholamines on the incidence of the preovulatory surge of of luteinizing hormone and ovulation in the rat: evidence for a necessary involvement of hypothalamic adrenaline in the normal or 'midnight' surge.

The preovulatory surge of luteinizing hormone reaches a maximum at 18.00 h on the day of pro-oestrus in female rats maintained with regular lighting from 06.00 to 20.00 h. This surge is initiated by a discharge of luteinizing hormone-releasing hormone into hypophysial portal blood. In this study, drugs which affect catecholamine-mediated neurotransmission were administered on the day of pro-oestrus and the effects on serum concentrations of luteinizing hormone and on subsequent ovulation were observed. alpha-Methyl-p-tyrosine, diethyldithiocarbamate and SKF 64139 inhibit catecholamine synthesis at the level of tyrosine hydroxylase, dopamine beta-hydroxylase and phenylethanolamine N-methyltransferase, respectively. Although alpha-methyl-p-tyrosine suppressed ovulation, it had a negligible effect on the incidence of the preovulatory surge. In contrast, the various treatments with diethyldithiocarbamate and SKF 64139 resulted in a minimal occurrence of the 18.00 h surge; at relatively low doses, however, these drugs frequently elicited a surge at 22.00 or 24.00 h which invariably resulted in ovulation. The failure of the surge after diethyldithiocarbamate or SKF 64139 was not associated with a loss of pituitary sensitivity to luteinizing hormone-releasing hormone. In terms of the hypothalamic concentration of dopamine, noradrenaline, adrenaline and 5-hydroxytryptamine at 18.00 h on pro-oestrus, the only common effect of diethyldithiocarbamate and SKF 64139, given in a dose which blocks the surge, was a severe depletion of adrenaline; alpha-methyl-p-tyrosine failed to produce this effect despite inducing a marked depression of dopamine and a moderate loss of noradrenaline. Neither the increase in hypothalamic dopamine after diethyldithiocarbamate, nor the alpha 2 receptor blocking properties of SKF 64139 appear to be relevant in this context since injections of L-dopa or piperoxane, an alpha 2 receptor antagonist, were without effect on the surge or ovulation. The failure of the surge after prazosin, an alpha 1 receptor antagonist, indicates that the function of adrenaline may be mediated postsynaptically by alpha 1 receptors. Clonidine, an alpha 2 receptor agonist which reduces the turnover rate of hypothalamic adrenaline, had effects of the surge and ovulation which were comparable to those of diethyldithiocarbamate and SKF 64139, the relatively low doses causing some of the surges to occur at 24.00 instead of 18.00 h and higher doses suppressing the surge at both times and thus preventing ovulation.(ABSTRACT TRUNCATED AT 400 WORDS)

Amines↗