Morphologic response of endometrium to a progesterone receptor antagonist, RU486, in monkeys

Fifteen castrate cynomolgus monkeys were assigned to one of two groups. All monkeys received estradiol (E2) capsules to mimic follicular phase E2 levels for development of a proliferative endometrium. In study I, three monkeys received RU486 on day 13 for 3 days; two others were given vehicle. On day 16, endometrium was removed. In study II, the remaining ten monkeys received progesterone capsules, beginning on day 12, to form a secretory endometrium. On day 29, six monkeys were given RU486 and four acted as controls. Endometrium was removed after 6, 18, and 32 hours. All tissue samples were prepared for light-microscopic evaluation. The results indicated that exogenous E2 alone stimulated intracellular stromal development; RU486 had weak progestational effect on proliferative endometrium, as evidenced by increase in the area occupied by glands; and RU486 had strong antiprogestational activity on secretory endometrium, illustrated by regression in the upper endometrial zones within 32 hours after a single injection of RU486.     

The effect of the antiprogestin mifepristone (RU 486) on maturation and in-vitro fertilization of human oocytes

The effect of RU 486 (mifepristone), a potent antiprogestin, on the in-vitro fertilization of human oocytes was investigated. In 40 normal volunteer women requesting laparoscopic sterilization, follicle aspiration for oocyte recovery was attempted 34 h after the injection of 5000 i.u. human chorionic gonadotrophin (hCG). Twenty women were allocated to receive 100 mg RU 486 orally 1 h before the hCG injection, the remaining 20 women acted as controls. There was no significant difference in the cleavage rate of the oocytes after fertilization in vitro between the two groups (56% and 66% respectively). Also, the morphological characteristics of the cleaving oocytes and the concentrations of oestradiol, progesterone and androstenedione in the follicular fluid of the leading follicle did not differ significantly between the two groups. Since RU 486 was detected in substantial amounts in the follicular fluid specimens, these results suggest that progesterone is not critical for the final stages of human oocyte maturation.     

The effect of the antiprogestin mifepristone (RU 486) on maturation and in-vitro fertilization of human oocytes

Summary. The effect of RU 486 (mifepristone), a potent antiprogestin, on the in-vitro fertilization of human oocytes was investigated. In 40 normal volunteer women requesting laparoscopic sterilization, follicle aspiration for oocyte recovery was attempted 34 h after the injection of 5000 i.u. human chorionic gonadotrophin (hCG). Twenty women were allocated to receive 100 mg RU 486 orally 1 h before the hCG injection, the remaining 20 women acted as controls. There was no significant difference in the cleavage rate of the ooctyes after fertilization in vitro between the two groups (56% and 66% respectively). Also, the morphological characteristics of the cleaving oocytes and the concentrations of oestradiol, progesterone and androstenedione in the follicular fluid of the leading follicle did not differ significantly between the two groups. Since RU 486 was detected in substantial amounts in the follicular fluid specimens, these results suggest that progesterone is not critical for the final stages of human oocyte maturation.     

The effect of progesterone and RU486 on progesterone production in the ovulatory process of rats

To clarify the autoregulatory mechanism of progesterone (P) production in the ovulatory process, we examined the ovarian concentration of P 46 hrs after PMSG and the effects of P and RU486 (RU) injected 4-2 hrs before hCG administration on the serum concentrations of P and estradiol (E2), and ovarian 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) activities in PMSG/hCG treated immature rats. The effect of RU on the number of ovulated ova was also studied. The ovarian concentration of P 46 hrs after PMSG was 0.96 +/- 0.03,ng/mg protein (mean +/- SEM). When P (100,mg/kg) was injected 2 hrs before hCG, ovarian 3 beta-HSD activities had significantly increased by 4 hrs after hCG. However, P at a dosage of 10 and 20,mg/kg had no effect on ovarian 3 beta-HSD activities. The administration of RU (20,mg/kg) 2 hrs before hCG significantly inhibited ovarian 3 beta-HSD activities measured 4 and 6 hrs after hCG (p less than 0.01 and p less than 0.05, respectively). In addition, the serum P concentration 4 hrs after hCG was significantly lower than that of the control (p less than 0.01). However, RU (20 mg/kg) in concomitant with hCG had no effect on ovarian 3 beta-HSD activities within 6 hrs after hCG. The suppression of ovarian 3 beta-HSD activities by RU was the concomitant reversed by the concomitant treatment with P (10 mg/kg). RU (10, 20, or 40 mg/kg) injected 2 hrs before hCG significantly reduced the number of ovulated ova (p less than 0.01, p less than 0.01 and p less than 0.01, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)     

Follicular maturation parameters associated with the failure of oocyte retrieval, fertilization, and cleavage in vitro

Follicular maturation parameters predictive of decreased success in human in vitro fertilization were evaluated in 104 gonadotropin-stimulated cycles. Falling estradiol (E2) levels correlated with decreased fertilization (1.9 oocyte/cycle) and cleavage (1.4 oocyte/cycle), when compared with cycles with increasing E2 (2.4 and 2.1 oocyte/cycle, respectively). Likewise, polyspermic fertilization was higher in the former group. Falling E2 levels after human chorionic gonadotropin (hCG) administration correlated with similar trends. Delaying hCG (24 to 96 hours) relative to the last dose of gonadotropin did not affect the total number of oocytes fertilized per cycle. However, polyspermic fertilization and cancellation rates were higher. Leading follicles growing beyond 2.3 cm (determined by ultrasound examination) were associated with decreased recovery, fertilization, and cleavage, and increased polyspermic fertilization. We conclude that ultrasonography and E2 monitoring can predict decreased fertilization and cleavage and should also be monitored after hCG administration.     

Pregnancy prevention by intravaginal delivery of a progesterone antagonist: RU486 tampon for menstrual induction and absorption

A progesterone antagonist (RU486) was administered to macaque monkeys by an intravaginal tampon for the achievement of both menstrual induction and absorption. The study was conducted in three parts: (1) postcoital treatment beginning 3 days after potential fertilization; (2) treatment on extended cycle days 34 to 37 after a "missed" menstrual flow after mating; and (3) luteal phase (cycle days 22 to 25) treatment in noncoital cycles. Our findings demonstrate pregnancy prevention and/or interruption when these treatments are employed.     

The physiological and clinical effects of progesterone inhibition with mifepristone (RU 486) in the second trimester

Summary. A double-blind placebo-controlled trial was performed in 20 primigravidae to assess the physiological and clinical effects of oral mifepristone on myometrial contractility and sensitivity in the second trimester. Ten women received 600 mg of oral mifepristone and 10 women a placebo 24 h before abortion was induced in both groups, with extra-amniotic PGE₂ instillation. Intrauterine pressure recordings demonstrated increased spontaneous uterine activity and increased sensitivity to PGE₂ and ergometrine, but no change in oxytocin sensitivity after mifepristone treatment. There were no significant differences in PGE or PGF metabolite concentrations in peripheral maternal plasma over the 24-h study period after treatment between the mifepristone and placebo groups. The mean induction abortion interval in the mifepristone group was 512 (SD 321) min compared with 1128 (SD 606) min in the placebo group (P≤0.02). The mechanism whereby mifepristone provokes enhanced uterine contractility and sensitivity to prostaglandins, with a reduction in abortion times, does not appear to be through endogenous production of PGE or PGF.     

The physiological and clinical effects of progesterone inhibition with mifepristone (RU 486) in the second trimester

A double-blind placebo-controlled trial was performed in 20 primigravidae to assess the physiological and clinical effects of oral mifepristone on myometrial contractility and sensitivity in the second trimester. Ten women received 600 mg of oral mifepristone and 10 women a placebo 24 h before abortion was induced in both groups, with extra-amniotic PGE2 instillation. Intrauterine pressure recordings demonstrated increased spontaneous uterine activity and increased sensitivity to PGE2 and ergometrine, but no change in oxytocin sensitivity after mifepristone treatment. There were no significant differences in PGE or PGF metabolite concentrations in peripheral maternal plasma over the 24-h study period after treatment between the mifepristone and placebo groups. The mean induction abortion interval in the mifepristone group was 512 (SD 321) min compared with 1128 (SD 606) min in the placebo group (P less than or equal to 0.02). The mechanism whereby mifepristone provokes enhanced uterine contractility and sensitivity to prostaglandins, with a reduction in abortion times, does not appear to be through endogenous production of PGE or PGF.     

Correlation between serum prolactin secretion patterns and the ability of fertilization and cleavage of human oocyte (effects of transient hyperprolactinemia on fertilization and cleavage of human oocyte)

To assess the effect of serum prolactin (PRL) on the fertilization and cleavage of oocytes, its secretion patterns in normoprolactinemic women (28 cases, 35 cycles) who had undergone IVF-ET because of bilateral tubal occlusion and/or severe oligospermia, and the relationship between its levels and the fertilization and cleavage rate were studied. Three serum PRL secretion patterns were observed. In the TIP-O (Transient Increase in serum PRL in the Ovulatory phase) group (7 cycles), the serum PRL was significantly increased in the ovulatory phase compared with the follicular phase. In the TIP-OL (TIP at the Ovulatory and mid-Luteal phase) group (11 cycle), it was significantly increased in both the ovulatory and the mid-Luteal phases, but in the NON-TIP group (17 cycles) there was no significant increase in any phase. The fertilization rate of the TIP-OL group (60.0%) was lower than that of the NON-TIP Group (89.5%) and the TIP-O group (87.5%). The cleavage rate of the TIP-O group (50.0%) was rather lower, and that of TIP-OL group (30.0%) was significantly (p less than 0.05) lower than that of the NON-TIP group (78.9%). These results suggest that transient increase in serum PRL may suppress the ability of fertilization and cleavage of human oocytes.     

The role of the progesterone antagonist,RU 486 (mifepristone), in preoperative cervical dilatation during the luteal phase. A randomized double-blind study

Endouterine surgery has become more common with the advent of operative hysteroscopy. It requires a preliminary dilatation of the cervix, which is usually achieved using rigid instruments. Cervical injury, uterine perforation, and other complications may occur in 2-5% of cervical dilatations for abortion and an unknown proportion in other uterine surgery. In as much as softening of the cervix has been observed in the 1st and 2nd trimesters of pregnancy after administration of sulprostone or of RU-486, a randomized double-blind study of RU-486 vs placebo was conducted to determine whether a progesterone antagonist could facilitate intrauterine surgery in the midluteal phase. 46 women undergoing hysterosurgery and laparoscopy under general anesthesia for idiopathic infertility were studied at the Antoine-Beclere Hospital in Clamart, France, between October 1987-October 1988. A placebo or 200 mg of RU=-486 was administered 2 days before surgery. For the 23 RU-486 and 23 placebo recipients respectively, the average ages were 31.2 and 32.5 years. 83% and 87% were consulting for primary infertility, and the average cycle day of surgery was 22.8 and 23.2. 6 women experienced spotting after RU-486 vs 1 in the placebo group. No undesirable secondary effects of complications were noted. No significant difference was seen in cervical consistency in the 2 groups. Average calibration evaluated with Hegar bougies was 5.15 for the RU-486 and 4.3 for the placebo group. A higher dose of RU-486 might have a greater cervical effect but would increase bleeding that would interfere with the hysteroscopy. No difference has been observed in the effect of RU- 486 administered in a single dose or divided into multiple doses. A change in the timing of RU-486 administration during the luteal phase and a shortening of the interval between RU-486 administration and surgery could be tested.     

人未成熟卵母细胞体外培养成熟、受精及胚胎移植

目的 应用卵母细胞体外成熟(IVM)技术帮助卵泡成熟障碍的不孕症患者获得妊娠及分娩。方法 接受未成熟卵IVM技术治疗者30例35个周期,其中多囊卵巢综合征14例,有卵巢过度刺激综合征病史6例,体外受精和胚胎移植(IVF-ET)周期中卵巢反应不良患者10例。设计卵巢刺激方案,采用含人成熟卵泡液的IVM培养液,建立未成熟卵母细胞的体外培养方法。结果 35个周期共取得未成熟卵母细胞203个,平均每个周期5．8个。培养后有156个卵母细胞排出第一极体,IVM率76．8％(156／203);在卵胞浆单精子显微注射(ICSI)12～18h后观察原核,正常受精率为76．9％(120／156);共有移植周期33个,获8例临床妊娠,妊娠率24％(8／33);有5例共7个婴儿出生。结论 IVM对一些卵泡发育和成熟障碍,特别是顽固性多囊卵巢综合征患者,是一种有效的辅助生育措施。人成熟卵泡液含有理想的自然促卵母细胞成熟的成分。     

Src家族激酶在卵母细胞减数分裂成熟和受精中的作用

Src家族激酶是一类非受体酪氨酸激酶。在卵母细胞成熟过程中,Src家族激酶能够促进卵母细胞恢复减数分裂。对海洋无脊椎动物和低等脊椎动物卵子受精过程的研究证明,Src 家族激酶与卵子活化过程中的Ca~(2+)释放有关；而在哺乳动物卵子中,对Src家族激酶的作用尚未形成一致结论。本文结合作者自己的工作,对目前Src家族激酶在减数分裂成熟和受精中的作用进行了总结和展望。     

Src家族激酶在卯母细胞减数分裂成熟和受精中的作用

Src家族激酶是一类非受体酪氨酸激酶。在卵母细胞成熟过程中，Src家族激酶能够促进卵母细胞恢复减数分裂。对海洋无脊椎动物和低等脊椎动物卵子受精过程的研究证明，Src家族激酶与卵子活化过程中的Ca^2＋释放有关；而在哺乳动物卵子中，对src家族激酶的作用尚未形成一致结论。本文结合作者自己的工作，对目Src家族激酶在减数分裂成熟和受精中的作用进行了总结和展望。     

Impact of the one-carbon metabolism on oocyte maturation,fertilization, embryo quality,and subsequent pregnancy

PurposeTo investigate impact of the one‐carbon metabolism (OCM) on oocyte maturity and embryo development.MethodsThis prospective study analyzed 18 women who agreed to participate. We measured the OCM biomarkers’ concentrations including Vitamin B12 (VB12), folic acid (FA), and homocysteine (Hcy) in serum and follicular fluid (FF), and assessed their correlation. We also evaluated the influence of such OCM biomarker concentrations in mono‐FF on oocyte maturation, fertilization, embryo quality, and consequent pregnancy after embryo transfers.ResultsAll biomarkers showed a high concentration variability in different follicles of each woman, but their mean levels correlated with the serum levels. Among the 106 collected oocytes, 92 were mature, 59 were fertilized, and 16 yielded good‐quality embryos. We performed 26 single embryo transfers, and 7 patients achieved clinical pregnancies. VB12 concentration (FF) was significantly lower in fertilized than unfertilized oocytes by univariate analysis. In multivariate logistic analysis, a significant correlation was found between FA concentration (FF) <14.25 ng/mL and good‐quality embryos and between Hcy concentration (FF) <4.9 nmol/mL and clinical pregnancy.ConclusionOCM in FF may affect fertilization, embryo quality, and clinical pregnancy.     

米非司酮对小鼠囊胚发育及超微结构的影响

目的:观察米非司酮对小鼠植入前胚胎在体内的生长发育及超微结构的影响。方法:雌性昆明小鼠,随机分为4组,每组15只:对照组(A组),米非司酮剂量1.9 mg/kg组(B组),5.6mg/kg组(C组)和16.8 mg/kg组(D组)。4组动物均按常规超排卵后合笼,在注射hCG后108h～110 h取囊胚观察,并用透射电镜检查4组囊胚超微结构变化。结果:A组和B组囊胚发育率无统计学差异(P>0.05),而C组和D组囊胚发育率显著减少(P<0.05)。随着米非司酮剂量增加,卵裂球表面粗颗粒增多,色泽深暗,卵周隙明显增大。超微结构观察显示,C和D组出现明显的膜、核、细胞器和染色质等超微结构病理改变。结论:米非司酮可损伤小鼠胚胎的超微结构,导致囊胚发育抑制,并呈现明显的剂量效应。     

The effects of platelet lysate on maturation,fertilization and embryo development of NMRI mouse oocytes at germinal vesicle stage

Purpose

Improving in vitro maturation could increase the rate of pregnancy from oocytes matured in vitro. Consequently, patients will be prevented from using gonadotropin with its related side effects. In this study, the maturation medium was enriched by platelet lysate (PL), then maturation and subsequent developments were monitored.

Methods

Oocytes at germinal vesicle stage with cumulus cells (cumulus–oocyte complex) and without cumulus cells (denuded oocytes) were obtained from mature female mice. The maturation medium was enriched by 5 and 10 % PL and 5 % PL + 5 % fetal bovine serum (FBS) as experimental groups; the control groups’ media consisted of 5 and 10 % FBS. After 18 h, the matured oocytes were collected and, after fertilization, subsequent development was monitored.

Results

The rates of maturation, fertilization and 2-cell embryo development for the denuded oocyte groups in experimental media 5 % PL and 5 % PL + 5 % FBS were significantly higher than those of the control groups (P < 0.05), while the results for the cumulus–oocyte complex groups were similar between the experimental groups and control groups.

Conclusions

The results of this study indicated that platelet lysate could improve the maturation rate in the absence of granulosa cells compared to media with FBS. This extract also had positive effects on fertilization and embryo development.

     

The function of Nucleoporin 37 on mouse oocyte maturation and preimplantation embryo development

Journal of Assisted Reproduction and Genetics - Nucleoporin 37 (NUP37) has been reported to activate the YAP-TEAD signaling, which is crucial for early embryo development. However, whether NUP37 is...     

Fertilization and cleavage of mouse oocytes exposed to the conditions of human oocyte retrieval for in vitro fertilization

Ova from two strains of mice (a hybrid-inbred strain, B6D2F1, and a random-bred strain, CD1) were shocked by exposure to environmental conditions possibly encountered by human oocytes retrieved for in vitro fertilization (IVF). Shocked and control mouse ova were fertilized in vitro in either simple or complex media and zygote development to morulae and blastocyst stages compared with that of zygotes fertilized in vivo. Development of the hybrid-inbred zygotes following fertilization in the simple media of shocked and control ova was essentially the same as for ova fertilized in vivo (84 +/- 6.6, 89 +/- 1.6, 87 +/- 6.0% to the 2-cell stage and 89 +/- 5.2, 94 +/- 2.3, 99 +/- 1.0% of two cells to blastocysts, respectively); development in the complex media also was the same following fertilization of shocked and control ova (80 +/- 8.7, 90 +/- 2.7% to two cells and 33 +/- 3.5, 35 +/- 4.5% of two cells to blastocysts, respectively) but lower than that of in vivo zygotes (92 +/- 4.6 to two cells, 58 +/- 4.7 two cells to blastocysts). In contrast, the fertilization and development in the simple media of shocked and control random-bred ova was lower and more variable (90 +/- 5.8, 67 +/- 11.1% to two cells and 17 +/- 8.3, 42 +/- 13.6% two cells to blastocysts, respectively) than the development of in vivo zygotes (96 +/- 1.5% to two cells, 51 +/- 5.5 two cells to blastocysts).(ABSTRACT TRUNCATED AT 250 WORDS)     

Relationship of human oocyte maturity,fertilization, and cleavage to follicular fluid prolactin and steroids

Follicular fluid (FF) levels of prolactin (PRL), estradiol (E₂), progesterone (P), and androstenedione (⁴A) were related to diversities in oocyte maturation, fertilization, and cleavage among oocytes obtained for in vitro fertilization. Follicles with mature stage I oocytes contained similar concentrations of PRL and E₂, more P, and less (⁴A) compared to follicles containing immature oocytes. Follicles with mature stage II oocytes had higher amounts of P and E₂ but similar levels of PRL and (⁴A) compared to follicles with mature stage I oocytes. Prolactin was lower and E₂ higher in FF associated with fertilizable oocytes than in those with unfertilized gametes. Follicles which yielded gametes that ultimately divided to more than four cells had the highest E₂ levels, while the lowest E₂ concentrations were associated with fertilized, uncleaved eggs. P, (⁴A), and PRL levels in FF were not related to oocyte cleavage. We conclude that (1) human menopausal gonadotropin (hMG) treatment does not necessarily lead to a synchrony or uniformity of preovulatory follicles, (2) maturation of oocytes is associated with rising levels of progesterone and decreasing levels of (⁴A) in FF, (3) an increased cleavage rate in vitro is associated with higher FF levels of E₂, and (4) high levels of PRL in FF are associated with unfertilized oocytes. These findings emphasize that differences in the hormonal milieu surrounding oocytes may have profound effects on the success of in vitro fertilization.Fogarty International (NIH, 5TWO3580) Fellow in Reproductive Biology and Endocrinology, on leave from Department of Obstetrics and Gynecology, Chaim Sheba Medical Center, TelHashomer, and Sackler School of Medicine, Tel-Aviv University, Tel-Aviv, Israel.