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PubMed1989

The infertility workup and diagnosis.

Infertility affects a significant number of couples. The traditional workup of the infertile couple involves performing a semen analysis, assessing ovulation with a postcoital test to evaluate sperm-mucus interaction, obtaining a hysterosalpingogram to assess uterine structure and tubal patency, and performing laparoscopy to rule out endometriosis and pelvic adhesive disease. In 1988 new techniques were added to our armamentarium, including the use of the sperm penetration assay (hamster test) to rule out occult male factors, the combination of multiple ovarian sonograms and luteinizing hormone surge detection kits to evaluate subtle problems of dysfolliculogenesis, labelled antiimmunoglobulins for the evaluation of sperm antibodies and diagnostic hysteroscopy for evaluating occult uterine pathology.

Fallopian Tube Diseases↗
PubMed1991

Detection of a soluble acrosome reaction-inducing factor, different from serum albumin, associated with the ovulated egg-cumulus complex.

Soluble extracts of the ovulated hamster egg-cumulus complex (ECC) were tested on capacitated sperm for activity in inducing the physiological acrosome reaction (AR). Evidence for occurrence of the physiological AR included enhanced sperm penetration of intact homologous zonae pellucidae as well as induction of AR in nonattached and in zona-bound sperm following a brief coincubation with test compound. Since hamster serum albumin, a major protein of hamster body fluids, also induces spontaneous ARs under certain conditions, it was used as one of the comparators for the acrosome reaction inducing factor (ARIF; Westrick et al., Biol Reprod 32 [Suppl 1]. 213, 1985) activity in the ECC. Sperm exposure to concentrations of the soluble ECC extract ranging from 0.04 to 0.2 mg protein/ml significantly increased penetration of salt-stored zonae by 36%, mean numbers of penetrating sperm by 90%, ARs in nonattached sperm by 65%, and ARs in zona-bound sperm by 102%. Hamster serum albumin added after completion of capacitation had no significant effect on these parameters. We conclude that 1) the ovulated ECC contains a soluble ARIF that augments zona-induced ARs and sperm penetration and 2) the ARIF is not serum albumin.

Acrosome↗
PubMed1999

Distribution of ganglioside GM3 in the rat ovary after gonadotropin stimulation.

Gangliosides are ubiquitous membrane components in mammalian cells and are suggested to play important roles in various cell functions, such as cell-cell recognition, differentiation and transmembrane signalling. Ovaries have been shown to contain GM3 as a major ganglioside. To study GM3 distribution during gonadotropin stimulation in the hypophysectomized rat ovary, ovarian sections and cultured granulosa cells were stained with specific monoclonal antibody against GM3. Interstitial cells of follicles of immature hypophysectomized rat ovary expressed ganglioside GM3. Theca cells of early antral follicles but not primary follicles expressed GM3. No granulosa cells of these follicles expressed GM3. When a surge dose of FSH/LH was injected, Graafian follicles were formed and GM3 expression was detected in granulosa cells of these follicles. After ovulation, cumulus cells kept expressing GM3 in the ampulla region of ovulated oviduct. The follicles did not show GM3 expression in their granulosa cells after an ovulatory dose of FSH/LH. At 48 h after in vitro culture with FSH/LH of granulosa cells from preantral follicles, GM3 was expressed to a detectable extent on the outer part of the granulosa layer. Finally, at 72 h after culture, all granulosa cells became positive to anti-GM3 antibody. These data suggest that the expression of ganglioside GM3 in the hypophysectomized rat ovary is spatiotemporally regulated by FSH/LH during follicular development and ovulation.

Animals↗
PubMed1982

[Studies on pregnancy specific beta 1-glycoprotein (SP1) by enzymeimmunoassay (author's transl)].

We developed a highly sensitive EIA for determination of SP1. The detection limit was 5ng/ml, and the correlation of EIA and RIA was extremely good (SAWAKI et al., 1980). Using this method, we found that SP1 was detected even in the serum at 12 days of ovulation. The concentration of SP1 in cord blood was on average 127ng/ml (24-540ng/ml). SP1 in newborn serum was 126ng/ml (22-355ng/ml) in 1st, and almost undetected in 4th day. SP1 in urine was 9.7ng/ml in the 5th week of pregnancy, 1,210ng/ml at peak in the 32th week, and in after delivery was 439ng/ml in 1st day and afterwards descended to 6ng/ml in 8th day. In 2 infertile women SP1 was detected in the sera of 14 days after ovulation. This suggests that SP1 measurment might be useful in the diagnosis of occult pregnancy. Three patients with throphoblastic disease were found to have regression curves for both HCG and SP1. The sera of 39 patients with uterine cervical carcinoma were examined to determine the occurrence of elevated SP1 levels. Ten of 27 patients before treatment had increased SP1 levels while only one of 12 patients had an increased SP1 level after treatment.

Choriocarcinoma↗
PubMed2002

The relationship between estrous behavioral score and time of ovulation in dairy cattle.

One of the major contributors to poor fertility of a dairy herd is ineffective detection of estrus. Recently, it has become evident that cow factors contribute largely to low detection rates. Until now, standing behavior has been the symptom used to determine the right moment for insemination. However, standing behavior is not observed in over 50% of the cows in estrus in a number of herds, and no data are available to relate the moment of ovulation to other estrous symptoms. Therefore, in the present study, cows were observed for a number of visual signs of estrus, twice per day for 30 min, and the moment of ovulation was estimated by transrectal ultrasonographic scanning of the ovaries. In this study, 100 cows were detected in estrus visually, using a scoring system. Standing behavior was observed in 50% of those cows, although there was more than one cow in estrus at a time in most (85%) cases. There was no correlation between follicular size and ovulation time or estrous behavior score. Levels of 305-d milk yield and parity were also not correlated with estrous behavior scores. However, a significant correlation (0.31) was found between detection score and day of ovulation. A retrospective analysis revealed that cows that ovulated 0 to 24 h after artificial insemination (AD) scored almost three times the number of estrous behavior points compared to the ones that ovulated 24 to 48 h after AI. Ovulation more than 48 h after AI resulted in pregnancy in only 15% of the cows.

Animals↗
PubMed1998

DNA strand breaks and phosphatidylserine redistribution in newly ovulated and cultured mouse and human oocytes: occurrence and relationship to apoptosis.

This study determined the occurrence of two molecular markers of apoptosis, chromosomal DNA strand breaks and oolemma phosphatidylserine redistribution, in >200 uninseminated and unfertilized human oocytes, and >800 newly ovulated and cultured mouse oocytes. DNA breaks were analysed by terminal deoxynucleotidyl transferase-mediated dUDP nick-end labelling (TUNEL) and phosphatidylserine by annexin V staining, with imaging by conventional epifluorescence and scanning laser confocal fluorescence microscopy. More than 300 intact and 500 fragmented mouse oocytes were examined at 24 h intervals during 6 days of culture in three different types of medium. For the human, 205 oocytes were examined at retrieval or at 24 h intervals during 7.5 days of culture in two types of medium. The perifollicular vascularity and the dissolved oxygen content of follicular fluid were determined for most of the follicles from which human oocytes were derived. The results demonstrate that TUNEL fluorescence of metaphase II (MII) chromosomes and annexin V staining of the oolemma in newly ovulated and cultured mouse and human oocytes are rare, and, when detected, are not spatially or temporally related. This finding also applied to mouse oocytes that fragmented during culture and exhibited morphological features that grossly resembled apoptotic body formation. In contrast, TUNEL but not annexin V staining occurred in the first polar body of a relatively high proportion of newly ovulated mouse oocytes, but was rarely detected in newly aspirated human oocytes. For the human, the occurrence of MII chromosomal TUNEL fluorescence was patient-specific and unrelated to perifollicular vascularity or dissolved oxygen content of the corresponding follicular fluid. The pattern of chromosomal TUNEL fluorescence observed in the first polar body and in the MII chromosomes of a very small number of mouse and human oocytes, especially after many days of culture, suggests that DNA strand breaks may not arise by apoptosis-associated endonuclease digestion. The results with these two markers suggest that it is premature to conclude that apoptosis occurs in ovulated oocytes or that such a mechanism is involved in the elimination or prevention of fertilization of oocytes with cytoplasmic or chromosomal defects.

Adult↗
PubMed1984

Efficacy of using vaginal conductivity as an indicator of the optimum time to breed in swine.

Two experiments were conducted to test the efficacy of using vaginal conductivity measurements to predict ovulation time in swine. In Exp. 1, 10 normally cycling 6-mo-old purebred Hampshire gilts were tested daily through two estrous cycles in order to characterize conductivity patterns. In all females, onset of behavioral estrus was verified by daily observation in the presence of a sexually active boar. Significant changes in vaginal conductivity by day of the cycle were detected in all females tested, with a high degree of variation among individuals. A 10% increase in conductivity occurred 12 to 24 h after the onset of estrus, which is coincident with the time normally associated with high fertility. In Exp. 2, 46 prepubertal crossbred and purebred Yorkshire gilts and 17 multiparous purebred Yorkshire and Duroc sows were assigned randomly to eight treatment groups. All females were checked twice daily for the onset of estrus and vaginal conductivity readings were taken from approximately 3 d before estrus until breeding. Females were bred by artificial insemination or handmated at times determined either by detection of estrus alone or when a 10% increase in conductivity readings occurred relative to the onset of estrus. Similar conception rates were achieved in animals bred after a 10% change in vaginal conductivity as compared with detection of estrus. These results indicate that vaginal conductivity measurements during estrus provided a reliable basis for determining the time to breed swine. However, there was no advantage associated with the use of vaginal conductivity.

Animals↗
PubMed2005

Form and function of the corpus luteum during the human menstrual cycle.

OBJECTIVE: To characterize the growth and regression of the corpus luteum (CL) during an interovulatory interval (IOI) using serial transvaginal ultrasonography. METHODS: Fifty healthy women of reproductive age with a history of regular menstrual cycles underwent daily transvaginal ultrasonography for one IOI. Measurements of luteal area and luteal numerical pixel value (NPV) were recorded each day after ovulation until the CL could no longer be detected. Blood was drawn every third day during the IOI to measure serum concentrations of progesterone and estradiol-17beta. RESULTS: Corpora lutea were of two morphological types: those with a central fluid-filled cavity (CFFC) (78%) and those without (22%). Eighty-eight percent of women exhibited a CL containing a CFFC 2 days after ovulation, followed by 34% 13 days after ovulation and 2% 27 days after ovulation. Luteal area, progesterone concentration and estradiol concentration increased for approximately the first 6 days following ovulation followed by a subsequent decline. Luteal NPV decreased from days 1 to 11 and increased during days 11-16. Changes in luteal area, NPV, progesterone and estradiol concentrations did not differ in women with two versus three waves of follicular development. CONCLUSIONS: Peak luteal function, as determined by maximum luteal area, progesterone concentration and estradiol concentration, is observed 6 days following ovulation. Luteal NPV is reflective of morphological and endocrinological changes in the CL. The development of a CFFC during luteinization is a normal physiological phenomenon. The CL can be detected, but is not functional, during the follicular phase of the menstrual cycle.

Adult↗
PubMed1977

Circulating levels of pregnancy-specific beta1-glycoprotein in early pregnancy.

Circulating levels of pregnancy-specific beta1-glycoprotein (SP1 or PSbetaG), luteinizing hormone (LH) and human chorionic gonadotrophin (HCG) were measured serially in 9 subjects immediately after conception. Ovulation occurred spontaneously in 3 subjects, or followed administration of clomiphene citrate (2 subjects) or bromocriptine (4 subjects). The timing of ovulation was determined by the appearance of the LH surge. Levels of HCG were detected 10 to 16 days, and SP1, 18 to 23 days after ovulation. These findings suggest that the measurement of plasma levels of SP1 may provide valuable additional biochemical evidence of pregnancy.

Chorionic Gonadotropin↗
PubMed1986

Induction of ovulation in the postpartum rhesus monkey: factors determining success in obtaining primate luteal tissue.

Ovulation induction in the postpartum rhesus monkey was attempted with the use of purified human pituitary gonadotropins for assessment of (1) whether this procedure could be used to provide a source of luteal tissue; (2) the extent of ovarian responsiveness to gonadotropic stimulation; (3) factors that might facilitate ovulation induction during the peurperium; and (4) factors that might be a contributory cause of induction failure. Twenty rhesus monkeys were treated with purified human follicle-stimulating hormone (hFSH) and human luteinizing hormone (hLH) twice daily, beginning on days 0, 10, or 20 postpartum, with or without prior administration of bromocriptine. Laparotomies were performed during the midluteal phase. Ovaries were examined, and all corpora lutea were removed. Neither the day of beginning the gonadotropin treatment nor bromocriptine administration had a significant effect on the success rate of ovulation induction, which averaged 60% overall. Inductions begun during July had a significantly (P less than 0.025) lower success rate than those started at other times of the year. Antibodies to hFSH and hLH were detected in serum from monkeys that had undergone ovulation induction. Antibodies to hFSH, but not hLH, were associated with significantly reduced induction success (P less than 0.05). Plasma estradiol rose in response to gonadotropin treatment, and the induced follicular phase averaged 13.6 +/- 0.7 days. In all animals judged to have ovulated, corpora lutea were observed at laparotomy, and plasma concentrations of progesterone were significantly elevated (13.8 +/- 3.8 ng/ml).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1998

Evaluation of timed insemination during summer heat stress in lactating dairy cattle.

We wished to compare the effect of summer heat stress on pregnancy rate in cows that were inseminated at a set interval associated with a synchronized ovulation vs those inseminated upon routine estrus detection. The study was carried out on a commercial dairy farm in Florida from May to September 1995. Lactating dairy cows were given PGF2 alpha (25 mg i.m.) at 30 + 3 d postpartum and randomly assigned to be inseminated at a set time (Timed group) or when estrus was detected (Control group). Cows in the Timed group were synchronized by sequential administration of Buserelin (8 micrograms i.m.) on Day 0 at 1600 h, PGF2 alpha (25 mg i.m.) on Day 7 at 1600 h and Buserelin (8 micrograms i.m.) on Day 9 at 1600 h. They were inseminated on Day 10 between 0800 and 0900 h (Day 9 + 16 h). Cows in the Control group were given PGF2 alpha at 57 + 3 d postpartum and inseminated when detected in estrus. Estrus detection or insemination rate for control insemination cows was 18.1 +/- 2.5% versus 100% for time inseminated cows (P < 0.01). Mean interval from PGF2 alpha to insemination was shorter for time inseminated cows (3 +/- 2.1 d < 35.5 +/- 1.9 d; P < 0.01). Pregnancy rate was greater for time inseminated cows (13.9 +/- 2.6 > 4.8 +/- 2.5%; P < 0.01) as was overall pregnancy rate by 120 d postpartum (27.0 +/- 3.6 > 16.5 +/- 3.5%; P < 0.05). Number of days open for cows conceiving by 120 d postpartum was less for time inseminated cows (77.6 +/- 3.8 < 90.0 +/- 4.2 d; P < 0.05), as was interval to first service (58.7 +/- 2.1 < 91.0 +/- 1.9 d; P < 0.01). Services per conception were greater for time inseminated cows (1.63 +/- 0.10 > 1.27 +/- 0.11; P < 0.05). The timed insemination program did improve group reproductive performance. However, the timed insemination program will not protect the embryo from temperature-induced embryonic mortality, but management limitations induced by heat stress on estrus detection are eliminated. An economical evaluation of the timed insemination program indicates an increase in net revenue per cow with implementation of timed insemination for first service during the summer months.

Animals↗
PubMed2001

Effect of short-term calf removal at three stages of a follicular wave on fate of a dominant follicle in postpartum beef cows.

The hypothesis that ovulation in response to short-term (48 h) calf removal (CR) is dependent on the developmental stage of the dominant follicle was tested in two studies. The objective of Exp. 1 was to characterize the fate of a dominant follicle following 48-h CR on d 2, 4, or 8 of a postpartum follicular wave. Ovaries of 61 beef cows were examined daily by transrectal ultrasonography starting at d 20 to 21 postpartum. Treatments were no CR (n = 14) and CR on d 2 (n = 12), 4 (n = 16), or 8 (n = 10) of first detected follicular wave. Percentage of cows that ovulated a dominant follicle following treatment was not different among groups (P = 0.62). Maximum size of dominant follicles was larger in cows that ovulated (P = 0.002) than in cows that did not ovulate. The objectives of Exp. 2 were 1) to determine whether a follicular wave could be synchronized in anestrous cows following injection of 1 mg of estradiol benzoate (EB) and 200 mg of progesterone (P4; EB + P4); 2) to characterize the fate of dominant follicles following 48-h CR at three stages of a synchronized follicular wave; and 3) to determine whether estrous cycles of normal length followed ovulation in cows pretreated with EB + P4. Ovaries of 50 anestrous beef cows were examined daily as in Exp. 1. Treatments were sesame oil (SO) injected (i.m.) on d 25 postpartum and no CR (n = 9); EB + P4 and no CR (n = 9); EB + P4 and CR on 6 (n = 12), 8 (n = 9), or 12 (n = 11) d after injection. The EB and P4 injections were given on d 25 postpartum. Variability in day of emergence of subsequent follicular waves was lower in cows receiving EB + P4 than in SO-injected cows (P < 0.05). The percentage of cows that ovulated was not different (P = 0.16), but CR increased the percentage of cows that ovulated when groups that received EB + P4 were compared to the EB + P4 group that did not have CR (53.1 vs 11.1%, respectively; P < 0.05). Maximum diameter of dominant follicles was larger (P = 0.05) in ovulatory follicles. The luteal phase was longer (P < 0.03) in cows receiving EB + P4 injection (10.6 +/- 1.2 d) than in cows receiving SO (4.4 +/- 2.2 d). In summary, the maximum size of ovulatory follicles was greater than that of nonovulatory follicles, the ovulatory response of postpartum anestrous cows was maintained through d 8 of a follicular wave, synchronization of follicular waves was accomplished in postpartum cows using EB + P4, and the subsequent luteal phase length was increased in animals that were administered EB + P4.

Anestrus↗
PubMed1975

Transitory changes in plasma progestins, estradiol, and luteinizing hormone approaching ovulation in the bovine.

Progestins, estradiol, and luteinizing hormone were measured by radioimmunoassay in bovine plasma samples collected from indwelling jugular catheters daily on days minus 6 to minus 4, every 6 h on days minus 3 and minus 2, and every 2 h from day minus 1 to ovulation. Least squares analyses characterized time trends within animals and interrelationships between these hormones. Plasma progestins decreased from 5.7 ng/ml at day minus 6 to .07 ng at the peak of luteinizing hormone. No preovulatory peripheral increase in plasma progestins was detected. Estradiol increased from 2 pg/ml at day minus 4 to 6 pg at minus 12 h and then increased abruptly to 7.4 pg at the peak of luteinizing hormone. This matter increase in estradiol was synchronous with the acute preovulatory surge of luteinizing hormone. Association was positive between estradiol and luteinizing hormone, but negative between progestins and estradiol. Estradiol decreased 50% by 5 h following the peak of luteinizing hormone with a return to base (2 pg) at 14 h. An increase of .1 ng luteinizing hormone/ml plasma per day was linear from day minus 6 to 8 h prior to the luteinizing hormone peak. Luteinizing hormone increased to a peak of 13.5 ng/ml and remained elevated for 10.5 h. These results are consistent with the hypothesis that an increase at proestrus in estradiol rather than progestins triggers the preovulatory surge of luteinizing hormone in the bovine.

Animals↗
PubMed1988

Natural and artificially induced ovulatory models related to lactation in the rat: role of prolactin.

The presence and the importance of a preovulatory prolactin (PRL) peak was determined in four, natural or artificially induced, ovulatory models related to lactation in the rat. Gonadotrophin peaks were determined in the afternoon preceding ovulation in four models: postpartum ovulation (PPO), ovulation after the lactational period (AL) (natural models), ovulation after litter removal at midlactation (ML), and ovulation in lactating rats (LR) (artificially induced models). In PPO, AL, and ML rats a preovulatory PRL surge was detected, showing that its presence is a common characteristic of ovulation in the rat. Bromocriptine inhibition of PRL levels in PPO and AL rats did not modify the percentage of rats which ovulated. In contrast, this treatment was able to significantly increase ovulation percentage in ML rats. Moreover, in LR rats strong dopaminergic inhibition of PRL levels, induced by pergolide, was necessary for ovulation to take place, but if pergolide-treated rats were injected with ovine PRL ovulation was completely inhibited. These data suggest that while a PRL surge seems to be always present in natural ovulatory models, it is not essential for ovulation to take place. On the other hand, in artificially induced ovulatory models, suppression of prolactinemia is able to induce ovulation or to increase the percentage of rats which ovulated. This effect of PRL on ovulation may be direct or indirect.

Animals↗
PubMed1998

Comparison of low-technology and high-technology monitoring of clomiphene citrate ovulation induction.

OBJECTIVE: To determine whether newer monitoring techniques, including urinary detection of the LH surge and vaginal ultrasound, offer an advantage over basal body temperature (BBT) charts alone in achieving pregnancy using clomiphene citrate (CC). DESIGN: Randomized clinical trial. SETTING: Infertility patients in an academic research environment. PATIENT(S): Forty-five women undergoing ovulation induction with CC. INTERVENTION(S): The women were assigned randomly to receive either low- or high-technology ovulation monitoring for a total of 3 ovulatory cycles. Both groups were followed with BBT charts. The high-technology group also was monitored with urinary LH kits and vaginal ultrasound. MAIN OUTCOME MEASURE(S): Cycle fecundity rates for each technique were compared statistically with use of life-table analysis. RESULT(S): Forty-five patients were studied during a total of 134 cycles. The overall cycle fecundity rate was 8%, 10% (8 of 81 cycles) for the low-technology monitoring group and 6% (3 of 53 cycles) for the high-technology monitoring group. These rates were not statistically significant when evaluated by Fisher's exact test (P = .53) or when using life-table analysis and a log-rank test (P = .48). CONCLUSION(S): These data suggest that, for initial attempts at ovulation induction with CC in unselected patients, high-technology monitoring of ovulation offers no increase in fecundity over low-technology monitoring.

Adult↗
PubMed1997

A study of early pregnancy factor activity in preimplantation.

PROBLEM AND METHOD: Early pregnancy factor (EPF), an Immunosuppressive substance, which appears in pregnant women's sera 48 h after fertilization, is a kind of pregnancy-specific protein. To determine whether the EPF activity could be a super early indicator of pregnancy, we used rosette inhibition assay to detect EPF activity in the sera, collected from 70 women 2-7 days after ovulation intending to conceive monitored by ultrasonography. Simultaneously we selected 40 non-pregnant sera and 12 early-pregnant sera as negative control and positive control, respectively. RESULTS: The results of this study demonstrated that EPF activity is detected in 35 women's sera out of 70 women within 2-7 days after ovulation, and 28 women out of the 35 were pregnant, which was known by follow-up, and 7 were not pregnant, possibly due to either false positive results or embryo loss because of preimplantation failure, thus causing no pregnancy. The other 35 out of 70 had no EPF activity and 34 of them were not pregnant, which was known by follow-up, but one case became pregnant, which was false negative result. Our study showed that diagnosis of the super early pregnancy could be made by detecting EPF activity in maternal serum within the time of preimplantation. The accuracy of pregnancy diagnosis by this method is 88.6%, with a false negative rate of 3.4% and a false positive rate of 17.1%. The beta-HCG level was measured from the above 70 women's sera in order to contrast EPF activity. All of the sera collected 2-6 days following ovulation indicated that there were lower beta-HCG values in very early pregnancy (> or = a5 mIU/ml). On the seventh day after ovulation, EPF activity was detected in 11 out of 15 sera with only 2 of them with a b-HCG level that reached or slightly surpassed that of the early pregnancy diagnosis (5 mIU/ml and 5.4 mIU/ml, respectively). This demonstrated that beta-HCG is not the earliest signal of pregnancy; otherwise the EPF activity is one that appears 2-6 days earlier than beta-HCG appears. We measured the progesterone level of the 48 sera from the 70 collected above within 2-7 days postovulation and found most of them reached the level of progesterone in the luteal phase (7.5-98.3 nmol/L). This indicated that ovulation had taken place in these women, which was in accordance with observations by ultrasonography. CONCLUSIONS: Our study showed that diagnosis (of 88.6%) of super early pregnancy could be made with an accuracy of 88.6% by detecting EPF activity in maternal serum within 2-days after ovulation. This offers a basis for pregnancy diagnosis for the women who attempt to terminate their pregnancy safely or who conceive unexpectedly, and it contributes to family-planning.

Adult↗
PubMed2006

A unique preovulatory expression pattern plays a key role in the physiological functions of BMP-15 in the mouse.

Mutations in the bone morphogenetic protein 15 (BMP-15) gene cause female infertility in the monoovulatory human and sheep; however, in the polyovulatory mouse, loss-of-function of BMP-15 results only in reduced ovulation rate. To elucidate the cause of these species-specific differences, we investigated the functional role of BMP-15 in the mouse ovary. Here, we found that the functional mature form of BMP-15 is barely detectable in the mouse oocytes until just before ovulation, when it is markedly increased. Further, we found that BMP-15 induces cumulus expansion in mouse cumulus-oocyte complexes. The oocyte culture medium from immature mice primed with pregnant mare serum gonadotropin followed by human chorionic gonadotropin also stimulated cumulus expansion, and this activity was attenuated by BMP-15 antibody. Interestingly, the oocyte culture medium from mice treated with pregnant mare serum gonadotropin alone had no effect. Moreover, BMP-15 stimulated the expression of EGF-like growth factors in cumulus cells as well as a series of molecules downstream of EGF-like growth factor signaling, including cyclooxygenase 2, hyaluronan synthase 2, tumor necrosis factor-stimulated gene 6, and pentraxin 3, all of which are necessary for normal cumulus expansion. An antagonist of the EGF receptor completely abolished the effect of BMP-15 in inducing cumulus expansion. These results are consistent with the phenotype of BMP-15-null mice, which exhibit normal folliculogenesis but have defects in the ovulation process. The species-specific differences in the phenotypes caused by BMP-15 mutations may thus be attributed to the temporal variations in the production of the mature form of BMP-15.

Animals↗
PubMed1978

Cell-to-cell communication and ovulation. A study of the cumulus-oocyte complex.

Cell-to-cell communication was characterized in cumulus-oocyte complexes from rat ovarian follicles before and after ovulation. Numerous, small gap junctional contacts were present between cumulus cells and oocytes before ovulation. The gap junction are formed on the oocyte surface by cumulus cell processes that transverse the zona pellucida and contact the oolemma. The entire cumulus mass was also connected by gap junctions via cumulus-cumulus interactions. In the hours preceding ovulation, the frequency of gap junctional contacts between cumulus cells and the oocyte was reduced, and the cumulus was disorganized. Electrophysiological measurements indicated that bidirectional ionic coupling was present between the cumulus and oocyte before ovulation. In addition, iontophoretically injected fluorescein dye was tranferred between the oocyte and cumulus cells. Examination of the extent of ionic coupling in cumulus-oocyte specimens before and after ovulation revealed that ionic coupling between the cumulus and oocyte progressively decreased as the time of ovulation approached. In postovulatory specimens, no coupling was detected. Although some proteolytic mechanism may be involved in the disintegration of the cumulus-oocyte complex, neither the cumulus cells nor the oocyte produced detectable levels of plasminogen activator, a protease which is synthesized by membrana granulosa cells. In summary, cell communication is a characterisitc feature of the cumulus-oocyte complex, and this communication is terminated near the time of ovulation. This temporal pattern of the termination of communication between the cumulus and the oocyte may indicate that communication provides a mechanism for regulating the maturation of the oocyte during follicular development before ovulation.

Animals↗