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PubMed1999

Incorporation of a GnRH agonist, leuprolide acetate, into regimens with exogenous gonadotropins to produce ovarian stimulation and ovulation in the nonpregnant squirrel monkey.

This study was designed to measure the effects of variations in the length of pretreatment with a GnRH agonist, leuprolide acetate (LA), on subsequent follicular development and ovulation. The hypothesis was that the duration of LA suppression of pituitary function does not adversely affect ovarian response to standardized ovulation induction protocols in squirrel monkeys. The first phase determined the dose and duration of LA needed to achieve a hypogonadal state. One of two groups received daily subcutaneous injections of 50 microg of LA. The other received a single injection of 175 microg of a depot suspension of LA. Sera were assayed for estradiol (E2) and progesterone (P). E2 and P levels increased 2- to 5-fold with peak levels on days 4 and 7, respectively. Suppression of steroid levels took 10 to 15 days in the LA-treated group. Depot-LA did not effectively suppress steroid production. After suppression, females receiving daily LA received five daily injections of hMG to stimulate follicular development. E2 and P increased in these animals. These results suggest that cycling squirrel monkeys have P-secreting capacity throughout the cycle. This may explain how the squirrel monkey is able to accommodate both a short (4-5 day) luteal phase of their 9 day cycle and implantation from 5 to 7 days after ovulation. A second study compared exogenous follicle stimulating hormone (FSH) to endogenous gonadotropins released as a response to LA in ovulation induction. Steroid production and hCG-induced ovulation were assessed. LA treatment was compared to a standard ovulation induction protocol by using a randomized cross-over measures design. There were no differences in E2 and P levels in response to dosages of either LA or hMG. The ovulatory response following LA treatment was not significantly greater than that using FSH. The number of animals with unovulated, large follicles was greater on the FSH protocol (12/18) compared to the LA protocol (3/18). Thus, a single injection of a depot preparation of LA is sufficient to stimulate follicular development and ovulation when followed by an hCG injection. Based on this observation and the data on unovulated large follicles, it is suggested that the ovary responds more readily to endogenous gonadotropins released by LA than to exogenous FSH.

Animals↗
PubMed1998

Analysis of aneuploidy rate in antral and ovulated mouse oocytes during female aging.

Two forms of oocytes termed SN (surrounded nucleolus) and NSN (nonsurrounded nucleolus) differing for the spatial distribution of nuclear and nucleolar-associated chromatin have been described within the antral compartment of the ovary of a number of mammals. The biological significance of these two kind of oocytes is as yet not completely clear. In previous studies we have shown that prior to ovulation, mouse SN oocytes isolated from the antral compartment, matured and fertilized in vitro have a far better meiotic and developmental competence than NSN oocytes. Immediately after ovulation SN and NSN oocytes remaining in the antral compartment do not develop beyond the 2-cell stage. To further examine the correlation between chromatin distribution and meiotic competence of mouse antral oocytes, in the present study we have analyzed chromosome segregation at the first meiotic division in antral (SN and NSN) and in ovulated oocytes. SN and NSN oocytes were isolated before (48 h post PMSG injection) or after (15 h post-hCG injection) ovulation from ovaries of females of increasing age, they were cultured in vitro to metaphase II, and their aneuploidy rate was examined. Comparison of data obtained before and after ovulation highlights two main points: 1. Following ovulation a statistically significant increase of aneuploidy is observed in antral oocytes in most age groups and it is attributable to SN oocytes. 2. The aneuploidy rate of ovulated oocytes does not increase during female aging. We have found a correlation between chromatin distribution, hormonal status, and the incidence of aneuploidy during the oocyte first meiotic division.

Aging↗
PubMed2000

Clomiphene citrate for ovulation induction in women with oligo-amenorrhoea.

BACKGROUND: The administration of clomiphene citrate is followed by an enhanced release of pituitary gonadotropins resulting in follicular recruitment. After the drug is stopped, there is continuing secretion of estradiol, selection of the dominant follicle and, in successful cases, ovulation. Clomiphene is indicated as the initial treatment in the majority of women with amenorrhoea and oligomennorhoea. In women with irregular ovulation it seems to re-establish typical frequency of ovulation. Its effectiveness in oligo-amenorrhoeic women was tested in a number of randomized controlled trials at that time. These trials form the basis for the following review. OBJECTIVES: Clomiphene citrate enhances the release of pituitary hormones, often resulting in ovulation. The objective of this review was to assess the effects of clomiphene citrate on ovulation and pregnancy in women with oligo-ovulatory subfertility. SEARCH STRATEGY: The Cochrane Subfertility Review Group specialised register of controlled trials was searched. SELECTION CRITERIA: Randomised trials of clomiphene compared with placebo or no treatment in women with oligo-ovulatory subfertility of at least 12 months duration. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. MAIN RESULTS: Four studies were included. They were all of crossover design. Since it was not possible to separate data from the first and second phases of these trials, the effect of clomiphene may be overestimated. Compared with placebo, clomiphene citrate was associated with increased ovulation. The odds ratio for high doses (50-250 milligrams per day) was 6.82 (95% confidence interval 3.92 to 11.85). This dropped to a non-significant odds ratio of 1.29 (95% confidence interval 0.48 to 3.49) with low doses (10 milligrams per day). Clomiphene citrate (all doses) was associated with an increased pregnancy rate per treatment cycle (odds ratio 3.41, 95% confidence interval 4.23 to 9.48). REVIEWER'S CONCLUSIONS: Clomiphene citrate (at doses between 50 to 250 milligrams per day) appears to be an effective method of inducing ovulation and improving fertility in oligo-ovulatory women. However adverse effects include possible ovarian cancer risk and risk of multiple pregnancy.

Amenorrhea↗
PubMed1978

Induced ovulation, development of the corpus luteum, and tubal transport in the striped skunk (Mephitis mephitis).

Ovulatory pattern, development of the corpus luteum (CL), and early tubal transport are described for 6 unmated and 19 mated striped skunks (mephitis mephitis) killed after known copulation times. Ovulation is induced by copulation and occurs around 42 hours (range = 40--50 hours) after first insemination. Seventeen of 19 females killed between 36 hours and 19 days postcoitus had ovulated. Two females killed at 36 hours had not ovulated; however, their follicles showed preovulatory changes and were significantly larger (p less than 0.01) than those of unmated females. Six unmated females, killed during and three weeks after the normal breeding season, had not ovulated. Ovarian events, such as preovulatory changes, ovulation, and development of the corpus luteum (CL), are described for animals killed 36 hours to 19 days post-coitus. Maximum CL size occurred at 11 days post-coitus and coincided with the onset of embryonic migration and enlargement. Embryonic development reflected individual variation in ovulation times. The first polar body was extruded at ovulation and the second at fertilization. Pronuclear stages were predominant at 42 to 48 hours and 3 to 8-cell stages at 72 to 96 hours; morulae (containing up to 95 cells) entered the uterus at seven days. Blastocysts were observed first at 11 days when embryonic spacing and enlargement began. Implantation occurred by 19 days.

Animals↗
PubMed1977

Environmental influence on ovulation and embryonic development in Rana pipiens.

Environmental effects on ovulation and embryogenesis in Rana pipiens were assessed using both freshly-captured fall animals and laboratory-conditioned females which had undergone vitellogenesis in the laboratory. Frogs in both categories were divided into two groups. Ovulation was hormonally induced in one group of females prior to cold exposure and in the second group of animals following an 8-week-period at 4 degrees C with an 8L 16D photoperiod. The incidence of both ovulation and normal embryonic development was increased following exposure of the animals to low temperatures and short daylength. Those animals which only partially ovulated prior to cold treatment did not respond to hormone injections following the period of cold exposure. Examination of the ovaries of these females revealed a much greater degree of oocyte resorption than was found in frogs whose initial ovulation was induced only after exposure to cold temperatures. The administration of ovulation-inducing hormones prior to artificial hibernation may thus have initiated a phase of oocyte resorption which progressed even at 4 degrees C. The incidence of ovulation was similar in wild-caught and laboratory-conditioned females, but eggs from the latter showed a much lower percentage of development to Shumway stage 20. This effect may have been related to differences in the environmental factors to whcih the two groups were exposed during oogenesis.

Animals↗
PubMed1982

In vitro ovulation of hamster oocytes depends on contraction of follicular smooth muscle cells.

Smooth muscle cells (SMC) of the hamster follicle contract prior to ovulation, and their contraction produces a V-shaped constriction in the base of the follicle. We have determined when basal constrictions first appear in follicles removed from females at various preovulatory times by examining stained, thick sections with a brightfield microscope. The time to ovulation was controlled by injecting the female with human chorionic gonadotropin (hCG). The shape of the follicle was classified as flat, small V, medium V, or large V depending on the size of the basal constriction. Twenty-nine out of 30 follicles removed 12 hours after the hCG injection had flat bases or small V-shaped constrictions. However, by 12.5 and 13.0 hours after hCG injection, 45 and 54%, respectively, of the follicles had medium and large V-shaped constrictions in their bases. Transmission electron microscopy was used to confirm that flat follicles had uncontracted SMC, follicles with small V's had uncontracted or slightly contracted SMC, and follicles with medium and large V's had highly contracted SMC. When 12 hour follicles were removed from females and cultured in vitro, ovulation did not occur. Microscopic examinations showed that SMC had not contracted in these follicles during culture. Some (36.6%) 12.5 hour follicles and most (69.9%) 13 hour follicles ovulated when cultured in vitro. The majority of those which failed to do so, did not have contracted SMCs. We interpret these data to mean that in hamsters follicular SMC begin to contract in some follicles between 12-12.5 hours after the hCG injection. If contraction has begun in vivo, then the ovary may be removed from the female and ovulation will occur during in vitro culture. If contraction has not begun in vivo, it will not do so using the in vitro conditions described in this study and ovulation will not occur. These observations are consistent with the idea that the contraction of follicular SMC is necessary for hamster ovulation, and suggest that either stretch or neurotransmitters may initiate SMC contraction in vivo.

Animals↗
PubMed1991

Fertilizing competency of multiple ovulated eggs in the domestic fowl (Gallus domesticus).

Fertilizing competency of multiple ovulated eggs in the domestic fowl was examined by fertilization in vitro and early development in culture. Normal laying hens (White Leghorn) were treated with 75 IU of PMSG for 7 days followed by injection of anterior pituitary extracts from chickens (CAPE). Ovulation began to occur 7.5 h after injection of CAPE. These hens ovulated 1-7 ova but some premature ovulation of GV stage ova were observed. In vitro fertilization of the multiple ovulated ova was examined by inseminating 10(6)-10(7) sperm onto the germinal disks in m-Ringer's solution. The gamete or zygote nuclei were detected by DNA specific fluorescence using DAPI (4',6'-diamidino-2-phenylindole) in the histological section prepared from the germinal disk. Process of fertilization was examined in the eggs incubated for 4 h after insemination in DMEM + liquid albumen at 41 degrees C under the atmosphere of 5% CO2 in air. Fertilization rate of the total multiple ovulated eggs was 55% (11/20), in which 90% (9/10) and 10% (1/10) in the eggs recovered 7.5-8.5 h and 9.0-9.5 h after CAPE injection were obtained, respectively. Normal pronuclei were formed in five eggs of those recovered 7.5-8.5 h after CAPE injection. Early development after fertilization in vitro was also examined by incubation for 12 h in DMEM + liquid albumen at 41 degrees C under the atmosphere of 5% CO2 in air. Although development in vitro was delayed compared to that in utero condition, normal development was observed in naturally and multiple ovulated eggs.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2005

Location of inclusion cysts in mouse ovaries in relation to age, pregnancy, and total ovulation number: implications for ovarian cancer?

Benign ovarian cysts are thought to be precursor lesions that differentiate and transform into carcinoma. With the aim of testing the hypothesis that increased ovulation number increases the frequency or number of ovarian cysts, the development and appearance of ovarian cysts was investigated in mice of differing ages and total lifetime ovulation number. High total ovulation number was induced by keeping mice in cages divided by a screen, with a male on one side and two females on the other side. Significantly more cysts were observed in animals subjected to incessant ovulation for 8 months and in 12 month breeding mice than in 3-month virgin mice or 1-month prepubertal animals. These cysts had the appearance of benign serous inclusion cysts. When cystic ovaries were serial sectioned, 47% of cysts had a connection to the ovarian hilus and potentially to the tubules of the rete ovarii, 31% were adjacent to the hilus, and 22% had an intra-ovarian location. A significant increase in intra-ovarian cysts was observed in the 8-month incessant ovulation group, implying that high ovulation number leads to ovarian surface invagination and inclusion cyst formation. In conclusion, ovarian inclusion cysts may be derived from more than one epithelial source, but incessant ovulation may increase the proportion derived from the ovarian surface epithelium. Because the cysts observed resembled human serous inclusion cysts these results have possible implications for epithelial ovarian carcinoma.

Aging↗
PubMed1997

Follicle and extrafollicular tissue interaction in 17alpha,20beta-dihydroxy-4-pregnen-3-one-stimulated ovulation and prostaglandin synthesis in the yellow perch (Perca flavescens) ovary.

Intact (attached to extrafollicular (EF) tissue) yellow perch follicles ovulate in vitro when stimulated with the steroid, 17alpha, 20beta-dihydroxy-4-pregnen-3-one (17,20-PG). However, follicles isolated from EF tissue will not ovulate under 17,20-PG stimulation. In the present study, the interaction of follicles and EF tissue in 17,20-PG-stimulated ovulation and prostaglandin synthesis was investigated by separating follicles from EF tissue at specific times during incubation. Following separation, the incubation of isolated follicles and EF tissue was continued. At 48 hr of incubation, isolated follicles were assayed for ovulation and the media of isolated follicle and EF tissue incubates were assayed by RIA for prostaglandin E (PGE) and prostaglandin F (PGF) levels. Steroid-stimulated follicles, isolated from EF tissue prior to 22 hr of incubation, ovulated less than 25%. However, there was a very large increase in the percentage ovulation in follicles separated from EF tissue from 24 to 32 hr. PGF levels increased significantly in 17,20-PG-stimulated follicles isolated from EF tissue after 22 hr. In contrast, mean PGE levels in steroid-stimulated follicle incubates appeared to be lower than control levels. Compared with controls, PGE and PGF levels were significantly lower in incubates of isolated EF tissue stimulated with 17,20-PG, and this did not appear to be related to the time of separation. The results demonstrate that follicles must interact with EF tissue for a specific length of time for ovulation and PGF synthesis to occur in isolated follicles. Further, the effects of 17,20-PG on prostaglandin synthesis appear to be specific for different tissue compartments.

Animals↗
PubMed1994

Di-(2-ethylhexyl) phthalate suppresses estradiol and ovulation in cycling rats.

Di-(2-ethylhexyl) phthalate (DEHP) is a known reproductive toxicant and a carcinogen in rodent animal models. DEHP may also be a reproductive toxicant in women. Its action in female animal models is undetermined, although its potential to target the ovary has recently been shown. We have identified the ovarian toxicity and target cells of DEHP in the female rat. Adult, regularly cycling Sprague-Dawley rats were dosed daily with 2 g/kg DEHP in corn oil by gavage for 1 to 12 days. Ovarian morphology and serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol, and progesterone levels were analyzed. DEHP exposure resulted in prolonged estrous cycles. Specifically, 35 of 42 DEHP-treated rats had 5- or 6-day cycles and only 7 of 42 had a 4-day cycle compared to 44 of 45 control rats with 4-day cycle lengths. DEHP treatment also suppressed or delayed ovulations by the first proestrus/estrus after metestrus-initiated dosing. Microscopic evaluation of the ovaries determined that 7 of 10 DEHP-exposed rats had not ovulated by vaginal estrus, whereas 13 of 13 control rats had ovulated by vaginal estrus. Thus, DEHP treatment significantly altered natural ovulation times. Preovulatory follicles were also quantitatively smaller in DEHP-exposed rats than in controls because the granulosa cells were smaller. The mean control rat preovulatory follicle granulosa cell area was 16 +/- 3 x 10(3) microns, whereas the mean DEHP-treated rat preovulatory follicle granulosa cell area was 12 +/- 4 x 10(3) microns. DEHP exposure significantly suppressed preovulatory follicle granulosa cell estradiol production over 8 days of exposure. Suppressed serum estradiol levels caused secondary increases in FSH levels and did not stimulate the LH surge necessary for ovulation. Consequently, ovulation did not occur in DEHP-treated rats, although DEHP-treated rats ovulated after treatment with human chorionic hormone. Vaginal lavage cytology from rats treated with DEHP did not detect the ovarian toxicity as shown by histology and hormone analysis. In summary, exposure to DEHP resulted in hypoestrogenic anovulatory cycles and polycystic ovaries in adult female rats.

Analysis of Variance↗
PubMed2000

Effects of polychlorinated dibenzofurans, biphenyls, and their mixture with dibenzo-p-dioxins on ovulation in the gonadotropin-primed immature rat: support for the toxic equivalency concept.

Previous studies have shown that 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), 1,2,3,7,8-pentachlorodibenzo-p-dioxin (PeCDD), and 1,2,3,4,7, 8-hexachlorodibenzo-p-dioxin (HxCDD), and their equipotent mixture block ovulation, reduce ovarian weight gain and alter preovulatory hormone levels in a similar manner. The objective of the current experiment was to investigate the effect of other structurally related compounds such as chlorinated furans and biphenyls on ovulation and related hormonal endpoints. The gonadotropin-primed immature female rat model was used to study the effect of 2,3,4,7, 8-pentachlorodibenzofuran (PeCDF), 3,3',4,4',5-pentachlorobiphenyl (PeCB), and 2,2',5,5' tetrachlorobiphenyl (TCB) and their mixture with polychlorinated dibenzo-p-dioxins (PCDDs) on ovulation. Rats were dosed on Day 23 of age at 0900 h with individual congeners (PeCDF, PeCB, TCB) or a mixture of five compounds, which included TCDD, PeCDD, HxCDD, in addition to PeCDF and PeCB. Equine choronic gonadotropin (eCG; 5 IU) was injected 24 h later to induce follicular development. Blood and ovaries were harvested, and ovarian weights determined at various times after eCG. Serum concentrations of 17beta-estradiol (E(2)), progesterone (P(4)), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) were determined by radioimmunoassay. At 72 h after injection of eCG, the number of ova shed was measured by irrigating the ova from oviducts. The slopes of the dose-responses for inhibition of ovulation generated by the individual PeCDF, PeCB, and/or their mixture with PCDDs were similar. PeCDF, PeCB, and the mixture increased serum concentrations of E(2) at 72 h after eCG injection, the day of expected ovulation; in contrast, serum P(4) and FSH were decreased at that same time point. Only the high doses of TCDD, PeCDF, and PeCB blocked LH and FSH surges at 58 h after eCG. The ovarian histology revealed that the effects of PeCDF, PeCB, and the mixture were very similar to those of PCDDs, consisting of ova in large preovulatory follicles and a lack of or reduced number of corpora lutea. Parallel dose-responses of the individual congeners (PeCDF and PeCB) and their equipotent mixture with PCDDs support the toxic equivalency (TEQ) concept for the blockage of ovulation. Thus, PCDDs, PCDFs, and PeCBs appear to block ovulation by the same or a very similar mechanism of action.

Animals↗
PubMed1978

Temporal relationships of estrogen, progesterone, and luteinizing hormone levels to ovulation in women and infrahuman primates.

These studies were undertaken to ascertain the interval between the estrogen and LH peaks and ovulation in women, rhesus monkeys, and baboons. Estrogen, progesterone, and LH were measured by RIA. Ovulation was documented by visual examination of the ovaries, histology of the corpora lutea, and recovery of ova. The data for human subjects was based on a group of 23 normal women scheduled for surgical sterilization. Blood was drawn daily between 8:30 and 10:30 P.M. beginning on day 10 of the cycle. Surgery was performed 1 to 5 days after the LH peak. The hormonal findings were correlated with the histology of the corpus luteum. The mean interval from the estrogen peak to ovulation was 34 hours, the interval from the estrogen peak to the LH peak was 24 hours, and that from the LH peak to ovulation was 9 hours. In 46 rhesus monkey cycles and in 53 baboon cycles diagnostic serial laparoscopic examinations were initiated following the estrogen peak and repeated every 24 hours until ovulation was confirmed. The mean interval between the estrogen peak and ovulation was 34 hours in the monkey and 41 hours in the baboon. The intervals from the estrogen peaks to the LH peaks were 12 hours in the monkey and 23 hours in the baboon. The intervals from LH peak to ovulation were 22 hours in the monkey and 18 hours in the baboon. Plasma progesterone levels were significantly increased prior to the LH peak in all three species.

Adult↗
PubMed1980

Blockade of ovulation and release of LH in the rat by electrochemical stimulation of the frontal lobe cortex.

The effect of stimulation of the frontal lobe cortex on the release of luteinizing hormone (LH) and ovulation was studied in female rats. Electrochemical stimulation (anodic DC) was applied through monopolar stainless-steel electrodes chronically implanted in the non-anesthetized freely-behaving animals bearing a plastic cannula inserted into the jugular vein for blood sampling. In rats, on the day of proestrus, stimulation (100 uA/30 sec) of the medial cortical surface in the superficial and deep layers of the medial precentral area and of the anterior cingulate area blocked ovulation in about 80% of the animals. A similar effect was seen when the stimulus was applied in the deep layers of the prelimbic and infralimbic areas. On the contrary, stimulation in the superficial layers of these latter two areas, as well as in the superficial and deep layers of the retrosplenial cortex, did not affect normal ovulation. The preovulatory discharge of LH was blocked in the animals which failed to ovulate. The degree of inhibition exerted by the anterior cingulate area and the prelimbic area on ovulation and LH surge was proportional to the amount of current applied. Stimulation of the anterior cingulate area also blocked the release of LH induced by the injection of progesterone into ovariectomized estrogen-primed rats. Furthermore, electrochemical stimulation of the anterior cingulate area inhibited the rise of LH in the serum induced by electrical stimulation of the medial preoptic area of ovariectomized estrogen-injected rats, but it failed to affect that resulting from electrical stimulation of the medial basal hypothalamus. On the other hand, stimulation of the lateral cortical surface and the ventral cortex of the frontal lobe on the day of proestrus affected normal ovulation and LH surge only when the stimulus was applied in the agranular insular area which also exhibited an inhibitory action. It is concluded that certain areas of the frontal lobe cortex related to limbic structures exert an inhibitory influence on ovulation and LH secretion.

Animals↗
PubMed1982

Cadmium: its effects on ovulation, egg transport and pregnancy in the rabbit.

The subcutaneous administration of cadmium chloride (CdCl2) 24 h before hCG-induced ovulation in rabbits sacrificed 14 h after induction of ovulation leads to a dose-dependent decrease in the number of oviductal eggs and the number of eggs shed. While a dose of 1.25 mg/kg CdCl2 imposed no effect, only 50-67% of the dogs treated with 2.5 or 5 mg/kg CdCl2 ovulated and of those ovulated eggs 35.6 and 45%, respectively, were found in the oviducts. At a dose of 7.5 mg/kg CdCl2, a higher proportion (57.1%) of the does failed to ovulate and only 16% of the eggs were recovered from the oviducts. Ligature of the uterotubal junction failed to increase the rate of eggs recovered from the oviducts. A sharp increase in serum progesterone (delta4 P) level within 2 h after induction of ovulation suggests that the reduction in the percentage of eggs recovered from the oviducts probably occurred due to altered fimbrial functions. On the other hand, in does treated with 5 mg/kg CdCl2 1 or 7 days before mating, pregnancy was interrupted in 60 and 75% of the does together with reduced delta4 P concentration and the conception rate (no. of implantations/CL) was significantly reduced. CdCl2 given 21 days before or 3 days after mating did not affect pregnancy and delta4 P concentration remained unaltered. Results suggest that the developing or mature follicles are sensitive to CdCl2 treatment. Once the CL is formed, the effect of CdCl is reduced. The receptivity of CdCl2-treated females indicates that probably the circulating estrogens were not affected and the ability to ovulate in response to hCG and mating reflects that, at least at the doses used, the hypothalamic-pituitary gonads functions are functional in the rabbit.

Animals↗
PubMed1986

Investigations on the control of in vitro spontaneous brook trout (Salvelinus fontinalis) ovulation.

The possible roles of follicular cyclooxygenase and cAMP in the control of in vitro spontaneous brook trout (Salvelinus fontinalis) ovulation were investigated. Brook trout oocytes that had undergone germinal vesicle breakdown and follicular separation in vivo, were incubated in vitro in the presence of indomethacin. At 3 or 30 microM, indomethacin significantly reduced the levels of PGF and PGE (measured by radioimmunoassay) in the incubation medium but did not inhibit spontaneous ovulation in vitro. Follicular cAMP levels were measured by a competitive protein binding assay, prior to and during spontaneous ovulation. cAMP levels were approximately 3.2 pmol/mg protein prior to incubation and did not fluctuate significantly from this value throughout the 24-hr incubation period. The phosphodiesterase inhibitor, 3-isobutyl-l-methyl-xanthine, significantly increased follicular cAMP levels at 1.0 and 0.1 mM. The combined results suggest that cyclooxygenase metabolites or a decrease in cAMP are not involved in the control of spontaneous brook trout ovulation in vitro. The in vitro effects of primaquine, a putative phospholipase mediator, were also investigated. At lower concentrations (0.1-0.5 mM), primaquine significantly enhanced ovulation above that observed in spontaneous controls. However, at 1.0 mM, primaquine inhibited spontaneous ovulation. Indomethacin at 3 or 30 microM did not block the stimulatory effect of primaquine observed at lower concentrations, indicating that cyclooxygenase metabolites are not involved in the stimulatory effect of primaquine on ovulation.

1-Methyl-3-isobutylxanthine↗
PubMed1989

In vitro ovulation, prostaglandin synthesis, and proteolysis in isolated ovarian components of yellow perch (Perca flavescens): effects of 17 alpha,20 beta-dihydroxy-4-pregnen-3-one and phorbol ester.

The present study investigated the effects of 17 alpha,20 beta-dihydroxy-4-pregnen-3-one (17,20 B-P) on (1) ovulation of isolated perch follicles in which the extrafollicular (EF) ovarian tissue had been removed; (2) prostaglandin F (PGF) and prostaglandin E (PGE) synthesis in EF tissue and intact (= follicles attached to EF tissue) and isolated follicles by radioimmunoassay and [14C]arachidonic acid incorporation; and (3) proteolysis in EF tissue and intact and isolated follicles by substrate sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In addition, the ovulatory and proteolytic effects of the phorbol ester, phorbol-12-myristate-13-acetate (PMA), and the calcium ionophore A23187 on 17,20B-P-stimulated follicles were also studied in the presence/absence of indomethacin (IM) and nordihydroguaiaretic acid (NDGA). Ultrastructural analyses revealed that the preparation of isolated follicles also removed the surface epithelium of the follicle. While 17,20B-P stimulated ovulation and an increase in PGF and PGE in incubates of intact perch follicles, it did not in incubates of isolated follicles. In contrast, PMA, A23187, and a combination of PMA and A23187 stimulated ovulation of these isolated follicles. PMA/A23187-induced ovulation could be blocked by NDGA but not IM, and two hydroxyeicosatetraenoic acids (11- and 15-HETEs) were capable of partially reversing the NDGA block. Incorporations with [14C]arachidonic acid revealed that the EF tissue had a significant potential to produce PGF; however, 17,20B-P did not stimulate an increase in PGF or PGE (measured by RIA) in incubates of EF tissue alone. In addition, neither ovulation nor an increase in prostaglandins was observed in cocultures of isolated follicles and EF tissue. One major protease (66 kDa) was observed in the medium during incubation of intact and isolated perch follicles. No protease activity was present in incubates of EF tissue alone. Protease activity in 17,20B-P-stimulated incubates of intact tissue was significantly higher than in steroid-stimulated incubates of isolated follicles. Protease activity increased in the medium during incubation with PMA or a combination of PMA and A23187. This activity was blocked by NDGA but not IM. The NDGA block was partially reversed by 11-HETE. The combined results suggest that there is an interaction of EF tissue and follicle that is necessary, particularly for the stimulation of ovulation and prostaglandin production. Further, the results with phorbol esters and NDGA suggest that the follicular control of ovulation in perch may involve protein kinase C acting through the production of a lipoxygenase product.

17-Hydroxycorticosteroids↗
PubMed1993

Evidence for 5-HT2 receptor involvement in the stimulation of preovulatory LH and prolactin release and ovulation in normal cycling rats.

The participation of serotonin (5-HT) in the control of LH secretion and ovulation has been reported in numerous pharmacological experiments, although the results have been contradictory. In this study to determine the physiological involvement of 5-HT2 receptors in the control of preovulatory LH and prolactin (PRL) release and ovulation, ketanserin (KET), a selective 5-HT2 receptor antagonist, was administered to normal cycling rats. Oral administration of KET at 10 mg/kg BW between 0900h and 1300h on the day of proestrus completely blocked ovulation, while the administration after 1500h did not. The ovulation was confirmed 2 days after treatment with KET. There was no consistent effect on the number of ova ovulated. Co-administration of KET with 5-methoxy-N,N-dimethyltryptamine (5-MeDMT), a 5-HT agonist, at a dose of 2 mg/kg BW ip reversed KET-induced inhibition of ovulation. The highest rate (89%) of reversal was obtained by co-administration at 1500h. KET administered at 1100h completely blocked the preovulatory surge of LH and PRL observed in control animals at 1800h. The present study suggests that 5-HT stimulates preovulatory LH and PRL surge and ovulation via 5-HT2 receptor under physiological conditions and that the effect of 5-HT depends on the critical period and the 24-hour periodicity.

Animals↗
PubMedDate not supplied

Ovulation in the perfused ovary in vitro: further evidence that estrogen is not required.

The effect of inhibition of estrogen synthesis on ovulation in rat ovaries perfused in vitro with medium without phenol red was examined. The addition of luteinizing hormone (LH, 0.1 microgram/mL) plus 3-isobutyl-1-methylxanthine (IBMX, 0.2 mM) to phenol red-free perfusion medium (M199 + 4% bovine serum albumin) induced ovulation. The number of ovulations was similar to that found in medium containing phenol red. There was a similar increase in estradiol (1, 3, 5 (10)-estratriene-3, 17 beta-diol) levels in the medium in both groups. The addition of 4-hydroxy-4-androstene-3, 17-dione (4-OH-A, 5 microM) to phenol red-free medium blocked the increase in estradiol levels induced by LH + IBMX, but did not prevent ovulation. There was no significant difference in the number of ovulations in the three groups. In conclusion, phenol red in the perfusion medium does not influence ovulation induced by LH + IBMX. Furthermore, an increase in estrogen is not required during the immediate preovulatory period for ovulation to occur.

1-Methyl-3-isobutylxanthine↗