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Comparison of transvaginal and transabdominal ultrasound for monitoring follicular development in an in-vitro fertilization programme.

Although nowadays most oocyte retrievals for in-vitro fertilization (IVF) are performed using a transvaginal, ultrasound-directed technique, monitoring of follicular development is still often performed via the abdominal route. On the bases of 106 sonographies carried out on 71 patients during ovulation monitoring within an IVF programme, we demonstrated that the number of follicles visualized endovaginally was significantly higher than that visualized by the abdominal route: n = 1124 versus n = 772 respectively, and that their size was significantly greater using the transvaginal approach: 20.8 mm versus 18.8 mm respectively, for the largest follicle. Consequently, if ovulation monitoring is carried out transvaginally, the follicular size criteria on which human chorionic gonadotrophin is administered must be reconsidered.

Abdomen↗

Does gonadotropin suppression result in follicular development in premature ovarian failure?

It was hypothesized that premature ovarian failure is a self-perpetuating problem and that suppression of the endogenously raised gonadotropin levels may allow synchronization of follicular growth, so that when the suppression is removed, follicular development may be stimulated by the rapid rise in follicle stimulating hormone (FSH) levels. Microgynon (ethinylestradiol, 30 micrograms and levonorgestrel, 150 micrograms) was administered to eight women with premature ovarian failure for 12 weeks. Serum samples were collected weekly for radioimmunoassay of FSH, luteinizing hormone (LH), estradiol, progesterone and inhibin. On stopping Microgynon, ovarian ultrasonography was carried out, in addition to blood sampling, to monitor any follicular growth. Both FSH and LH levels declined to the normal follicular-phase range after 5 weeks of Microgynon treatment (mean +/- SD: FSH, 6.5 +/- 1.2 IU/l; LH, 8.9 +/- 1.6 IU/l). Both hormone levels rose rapidly after stopping treatment and were above the normal follicular-phase range by the 1st week. FSH reached pretreatment levels by 3 weeks, but LH did not reach the previously high levels until 7 weeks after stopping Microgynon administration. Estradiol and immunoreactive inhibin levels in serum were undetectable throughout the study. No follicular growth was seen on ultrasound scanning. Therefore, gonadotropin suppression in premature ovarian failure does not result in a resumption of follicular activity.

Adult↗

Follicular development and a single ovulation induced with pulsatile administration of Gn-RH in anovulatory women: studies of hormonal analysis and follicular sonometry.

The method of pulsatile administration of gonadotropin-releasing hormone (Gn-RH) has been proven as a useful means for induction of ovulation in anovulatory women. In our series of clinical trials, 23 out of 29 anovulatory patients ovulated with pulsatile administration of Gn-RH. Seven patients who ovulated volunteered for the present study with daily hormonal analysis and follicular sonometory . Two patients have oligomenorrhea, 3 patients secondary amenorrhea-1st grade (the sole administration of gestagen required for withdrawal bleeding) and the remaining 2 patients secondary amenorrhea-2nd grade (the combined administration of estrogen and gestagen required for withdrawal bleeding). A diagnosis of hyperprolactinemia was made for one patient with secondary amenorrhea-1st grade. Pulsatile administration of Gn-RH was performed by the use of a self-administered infuser . The infuser was connected to an i.v. indwelling catheter via a specially designed blood backflow eliminater . Five micrograms or less of Gn-RH was given every 2 hr from 07:00 to 23:00 hr daily. Five patients received HCG during the preovulatory period. In one patient, a short term treatment of HMG was added to Gn-RH treatment. Follicular sonometry revealed the development of a single dominant follicle which reached between 20 and 28 mm (23.7 +/- 0.12 mm, mean +/- S.E.) in diameter at the preovulatory period. Disappearance of a dominant follicle was recognized in the early luteal phase. Characteristic increases in estradiol were recognized concomitantly with the development of a dominant follicle. Progesterone levels after ovulation were within the limits of its normal "luteal phase" rise. The present data suggest that pulsatile administration of low dose Gn-RH with nocturnal interruption of treatment is effective for normal progress of follicular development in various types of anovulatory patients, culminating in single ovulation. This paper includes the discussion on our method which may be responsible for a high success rate of ovulation induction.

Adult↗

Aromatase activity in human granulosa cells during follicular development and the modulation by follicle-stimulating hormone and insulin.

Granulosa cells were isolated from follicles that ranged in size from 0.4 to 2.0 cm in diameter, obtained from patients during the follicular phase of the menstrual cycle. Aromatase activity, assessed by the release of tritiated water from [1 beta-3H]testosterone, was undetectable in follicles with a diameter of less than 1.0 cm; thereafter, there was a direct correlation between aromatase activity and follicular diameter. When cultured in the presence of 200 ng of National Institutes of Health follicle-stimulating hormone-15 per milliliter, aromatase activity was stimulated in cells isolated from all sizes of follicles from 0.4 to 1.5 cm. Insulin (500 ng/ml) further augmented follicle-stimulating hormone (FSH)-induced aromatase activity, as evidenced both by an increase in 17 beta-estradiol production and by the release of tritiated water from [1 beta-3H]testosterone by granulosa cells. This study further delineates the role of FSH in estrogen production during human follicular development and suggests that insulin or insulin-like growth factors may play a role in modifying the FSH-dependent cellular differentiation of human granulosa cells.

Adult↗

Evidence for the action of bovine follicular fluid factor(s) other than inhibin in suppressing follicular development and delaying oestrus in heifers.

The aim of this study was to investigate the importance of inhibin in the delay in return to oestrus in heifers induced by steroid-stripped bovine follicular fluid (bFF). Oestrous activity was synchronized in 18 Hereford x Friesian heifers with two injections of prostaglandin (PG) 12 days apart. At the time of the second PG injection (time 0), the animals were assigned at random to one of three experimental groups and received i.v. injections of 20 ml saline (controls, n = 6), whole bFF (FF group, n = 6) or bFF in which the bioactive inhibin content had been reduced by > 95% by immunoaffinity chromatography (-INH group, n = 6; inhibin content approximately 0.8 ml whole bFF) every 8 h for 2 days. In a dose-response study, 2.5 ml whole bFF was insufficient to delay oestrus consistently following a similar synchronization regimen. Blood samples were taken every 8 h, initially before each injection and then subsequently for a further 9 days for hormone analysis. Animals were observed every 8 h throughout the experiment for signs of behavioural oestrus. The ovaries of all animals were examined using real-time ultrasonography about 30 h after the second PG injection. Treatment failed to suppress peripheral follicle-stimulating hormone (FSH) concentrations, although a significant increase was observed in both treatment groups after cessation of injections. Progesterone concentrations fell immediately after the second PG injection in all animals and remained below minimum detectable concentrations in all treated animals for the remainder of the experiment. In control animals, progesterone rose above minimum detectable concentrations by day 6 and continued to rise until the end of the experiment. Analysis of samples taken from treated animals several days after observed oestrus revealed that all had apparently ovulated. Mean daily luteinizing hormone (LH) concentrations did not differ between treatment groups before ovulation, but after ovulation, mean daily LH was significantly reduced in control animals as progesterone concentrations rose. Follicular development, as assessed by the mean antral diameter of the largest follicle on a pair of ovaries at ultrasound examination, was significantly suppressed in treated animals compared with controls (P < 0.01) and there was no significant difference (P = 0.397) between the two treatment groups. Control animals displayed oestrus 68 h (+/- 8 SEM) after the second PG injection, but oestrus was delayed in treated animals to 186h +/- 5 (FF group) and 191 h +/- 6 (-INH group).

Analysis of Variance↗

Suppression of basal secretion of FSH inhibits follicular development and maturation during the oestrous cycle of the rat.

The role of basal FSH secretion during the rat oestrous cycle in regulating ovulation was examined by suppressing FSH secretion using charcoal-treated porcine follicular fluid (pFF). Although 0.5 ml pFF given at 05.00 and 11.00 h on the day of pro-oestrus had no effect on ovulation, 0.5 ml pFF given five times at 6-h intervals from 11.00 h on dioestrus to 11.00 h on pro-oestrus completely eliminated ovulation on the morning of the next oestrus. When 0.25 ml pFF was given on the same schedule, all animals ovulated a significantly decreased number of oocytes (9.0 +/- 0.8) at the next oestrus. During the period of pFF treatment, the number of follicles capable of ovulating in response to human chorionic gonadotrophin (hCG) decreased (7.6 +/- 0.7 at 05.00 h on pro-oestrus) and plasma levels of oestradiol showed a peak level 6 h later than in controls treated with 0.5 ml steroid-free porcine serum. Treatment with pFF suppressed plasma FSH concentrations in a dose-dependent manner, although plasma LH was inhibited irregularly. Supplementary administration of LH throughout the treatment period and 0.5 ml pFF resulted in ovulation of one to three oocytes in response to hCG in only three out of ten animals. These results suggest that basal secretion of FSH during the rat oestrous cycle plays an essential role in follicular development and maturation towards ovulation, and that the levels of FSH secretion may contribute to the maturation of normal numbers of follicles for ovulation.

Animals↗

Effects of functional peripheral sympathetic denervation induced by guanethidine on follicular development and ovulation of the adult female guinea pig.

The present study investigates the effects of functional sympathetic peripheral denervation induced by guanethidine sulphate (GTD) to adult female guinea pigs in the follicular (FPh) or luteal phases (LPh) on their oestrous cyclicity and ovulation. No differences were observed in oestrous cyclicity or the average number of corpora lutea present in the ovaries between the control and denervated animals. Guanethidine sulphate administration resulted in a significant decrease in ovarian norepinephrine content, higher for the left ovary than for the right one. Serum oestrogen and progesterone concentrations, the mean of follicles, and its diameter were different, depending on the oestral cycle in which the treatment was performed. These results suggest that in adult normal female guinea pigs, ovarian innervation participates in the regulation of follicular development in an inhibitory way.

Animals↗

Effects of unilateral ovariectomy on follicular development and ovulation in cattle.

In a study of 4 cyclic dry cows (Trial I) and 6 cyclic puberal heifers (Trial II), unilateral ovariectomy increased the number of ovulatory follicles, did not alter the hormone profile, cycle length or the number of follicular waves. Ovarian follicular development in all 4 cows was monitored daily using transrectal ultrasonography until the day of ovulation, during which period daily blood samples were also taken from the tail vein for determination of plasma FSH, LH and P4 concentrations. Unilateral ovariectomy was performed on the day after ovulation and ovarian activity was again monitored daily (ultrasonography and blood sampling for FSH, LH and P4) for 2 consecutive cycles (8 cycles in all). Estrus in all 6 heifers was synchronized using 2 injections of PGF2 alpha given 12 d apart. Similarly, ovarian activity in the 6 puberal heifers was monitored daily using ultrasonography and blood sampling for 1 complete control cycle. Following estrus and ovulation the left ovary was removed in all the animals, and thereafter 1 complete cycle was followed. Mean cycle length, FSH, LH and P4 concentrations before and after unilateral ovariectomy were compared using paired sample t-test. The results show that unilateral ovariectomy neither altered the cycle length nor the number of follicular waves in the cows, but it increased the number of ovulatory follicles (2 follicles developed and ovulated in 6 of the 8 cycles). The mean diameter of the largest follicle was 16.1 +/- 0.9 mm and the second largest 12.5 +/- 0.9 mm. No significant (P > 0.05) differences were observed in FSH (0.72 +/- 0.09 vs 0.71 +/- 0.07), LH (0.42 +/- 0.1 vs 0.37 +/- 0.07) and P4 (2.8 +/- 0.6 vs 2.6 +/- 0.4) levels before and after unilateral ovariectomy. Of the 6 heifers, 5 had 2 waves and 1 heifer had 3 waves of follicular growth during the control cycle, and this pattern did not change after the procedure. Mean cycle length (20.7 +/- 0.9 vs 21 +/- 0.9) did not differ before and after unilateral ovariectomy, and 4 of the 6 heifers ovulated twin follicles following ovariectomy. The mean diameter of the largest follicle was 14.5 +/- 0.7 mm and second largest measured 12.1 +/- 0.8 mm. No significant (P > 0.05) differences were observed in FSH (0.16 +/- 0.09 vs 0.21 +/- 0.07), LH (0.11 +/- 0.1 vs 0.15 +/- 0.07) and P4 levels (3.6 +/- 0.26 vs 3.8 +/- 0.29) before and after unilateral ovariectomy. Based on these results, we conclude that unilateral ovariectomy is an ideal method for obtaining twin ovulations in cows and heifers.

Animals↗

Co-expression of messenger ribonucleic acids encoding IGF-I, IGF-II, type I and II IGF receptors and IGF-binding proteins (IGFBP-1 to -6) during follicular development in the ovary of seasonally anoestrous ewes.

In recent years, it has become apparent that components of the insulin-like growth factor (IGF) system are involved in the regulation of ovarian follicular development in sheep. The majority of previous studies have concentrated on investigating only a select few components and not the whole system. The aim of the present study was to use five seasonally anoestrous ewes to investigate the expression of mRNA encoding all 10 components of the sheep IGF system among various-sized follicles within the ovary, using sheep-specific ribonucleotide probes and in situ hybridisation. IGF-I mRNA expression was low and did not vary with follicle size. IGF-II mRNA expression was significantly higher (P < 0.05) in small follicles compared to large follicles. Both IGF receptors had significantly higher (P < 0.05) levels of mRNA expression in small follicles, with the type I receptor being expressed to a slightly greater extent than the type II receptor. IGFBP-2, -3, -4 and -5 gene expression followed a similar pattern to IGF-II and the IGF receptors, whereby expression decreased with increasing follicle size. Similar to IGF-I, IGFBP-6 mRNA expression showed little variation with follicle size. IGFBP-1 mRNA expression was observed at low and constant levels, albeit in small and medium-sized follicles only. These data demonstrate that all of the components of the IGF system are produced in the ovine follicle, and for some of the components, their gene expression varied with stage of follicle development. This study further emphasises the importance of IGF-II as the major IGF in the autocrine and paracrine regulation of follicle development in sheep.

Anestrus↗

Gonadotrophin secretion in the second half of the menstrual cycle: a comparison of women with normal cycles, luteal phase defects and disturbed follicular development.

In order to study the relationship between episodic gonadotrophin secretion and alterations of ovarian hormone secretion, we examined women with normal menstrual cycles (n = 26), luteal phase defects (n = 10) or disturbed follicular oestradiol secretion (n = 8) as established by daily (except weekends) determinations of oestradiol and progesterone. Pulsatile gonadotrophin secretion was studied during the luteal phase or the second half of the menstrual cycle sampling at 15 min intervals for 12 h. LH and FSH mean concentrations and LH pulse frequency were significantly (P less than 0.01) increased in the group with disturbed follicular development in the presence of decreased oestradiol (E2) and progesterone (P4) levels. In women with luteal phase defects mean LH and FSH concentrations and pulsatile LH secretion showed a nonsignificant trend to lower values in the presence of significantly decreased P4 concentrations during the luteal phase.

Estradiol↗

Quantitative investigation of the annual pattern of follicular development in the nine-banded armadillo (Dasypus novemcinctus).

Mature, non-pregnant, female armadillos (Dasypus novemcinctus) were killed throughout the year (3-6/month, N = 54) and both ovaries were examined for follicular development. All normal and atretic follicles > 358 micrometers were counte. Total number of normal follicles remained constant from January to June but decreased by 50% in the remaining months except in October and November. Follicles > 978 micrometer (those likely to be ovulated) were found during each month but more (> 3/ovary) were present during April, May, June and October. Atretic follicles remained constant (4-6) except in July and August when the number (10-12) was doubled. A single CL was present in 2/4, 6/6, 1/6, 1/3 and 2/3 animals in July, August, October, November and December, respectively. These data confirm that armadillos ovulate in July and suggest that ovulation may also occur in some animals in late October or November.

Animals↗

Follicular development and ovulation in macrophage colony-stimulating factor-deficient mice homozygous for the osteopetrosis (op) mutation.

To clarify the role of macrophages and macrophage colony-stimulating factor (M-CSF) in follicular development and ovulation, the processes of folliculogenesis and ovulation, numerical changes in macrophages, and proliferative capacity of granulosa cells were examined in op/op mice before or after daily M-CSF administration. The natural estrous cycle was determined daily by means of vaginal smears. The number of ovulated ova in both fallopian tubes was significantly smaller in op/op mice than in normal littermates. Such ova markedly increased in number after daily M-CSF administration. The numbers of both antral and mature follicles in the proestrous ovary were markedly lower in op/op mice than in the controls and increased after daily M-CSF administration. Flash-labeling with [3H]thymidine showed that the proliferative capacity of granulosa cells in antral follicles was reduced in op/op mice but elevated after daily M-CSF administration. Numbers of granulosa cells and macrophages in the antral follicles were significantly decreased in op/op mice but were increased after M-CSF treatment. All these data provide evidence that macrophages are implicated in the process of folliculogenesis and ovulation.

Animals↗

The effect of feeding diets high in quickly degradable nitrogen on follicular development and embryo growth in lactating Holstein dairy cows.

This study investigated the effect on follicular and embryo development of increasing quickly degradable nitrogen (QDN) intake in lactating Holstein dairy cows. Forty mature post-partum cows were fed one of two diets for a minimum of 10 weeks, starting 10 days before first insemination. The Control diet was a high production dairy ration. The High QDN diet comprised the Control ration plus 250 g urea/head/day. Both diets were formulated to ensure that the energy requirements of the cows were satisfied. The High QDN treatment resulted in a significant increase in milk urea, plasma urea and plasma ammonia concentrations. The highest plasma urea (8.2 mmol/l) and ammonia concentrations (120 micromol/l) were recorded within 7 days of the urea supplement being added to the diet. There was no effect of diet on plasma progesterone or glucose concentration. There was also no significant effect of treatment on follicular development or embryo growth. The results from this study suggest that the lactating cow can adapt to increased intakes of QDN if the increase starts at least 10 days before insemination.

Ammonia↗

Significance of oestradiol for follicular development in hypogonadotrophic immature rats treated with FSH and hCG.

Administration of hCG (0.5 i.u.) to immature female rats that were made hypogonadotrophic by injections of an LHRH-antagonist at 22, 24, 26 and 28 days of age and in which follicular development was induced by FSH (10 i.u., given from day 26 until day 29) stimulates follicles to grow to their preovulatory size with a low amount of atresia. To investigate whether this effect of hCG on follicular growth is due to stimulation of oestradiol production, intraovarian concentrations of oestradiol were suppressed by an aromatase inhibitor and oestradiol action was blocked by an oestrogen antagonist; both of these were administered from day 25 until day 29. By day 30 this treatment had resulted in an increase in the percentage of atretic follicles. In turn, oestradiol benzoate (100 micrograms or 1000 micrograms day-1), given from day 26 until day 29 in rats treated with 10 i.u. FSH, resulted in (1) an increase in the total number of antral follicles, (2) an increase in follicular size and (3) a decrease in the percentage of atretic follicles. Although the administration of 1000 micrograms oestradiol benzoate can mimic the effects of 0.5 i.u. hCG on follicular growth to a large extent, the number of follicles with a diameter > 575 microns in rats treated with FSH plus oestrogen was smaller than that in rats treated with FSH plus hCG (10 +/- 2 and 24 +/- 2, respectively). Furthermore, a single injection of 10 i.u. hCG given on day 29 was used to induce ovulation: fewer ova were found in rats treated with FSH plus 1000 micrograms oestrogen than in rats treated with FSH plus hCG (15.2 +/- 2.1 and 46.5 +/- 1.6, respectively). From these studies, it is concluded that the effects of hCG on follicular growth and atresia can be largely attributed to the mitotic and anti-atretic effect of oestradiol. However, part of the effect of hCG cannot be explained on the basis of oestradiol action.

Animals↗

Follicular development and reproductive endocrinology during and after superovulation in heifers and mature cows displaying contrasting superovulatory responses.

To understand the causes for poor response to superovulation in mature cows of high genetic potential, endocrine and follicular events during and after superovulation were compared in heifers (<2 yr old) yielding large numbers of embryos and cows (9 to 13 yr old) known to be poor embryo donors. Follicular development was monitored by daily ultrasonography. Blood samples were taken 2 to 3 times a day for the measurements of P4, E2, FSH and LH by RIA. Intensive blood collections at 15-min intervals for 6 h were also performed during preovulatory and luteal phases. The number of embryos produced in the heifers (15.2 +/- 2; mean +/- SEM) and the cows (0.6 +/- 0.4), was similar to the number of ovulatory follicles derived from ultrasonographic observations in the heifers (16.2 +/- 3.7), but not in the cows (7.8 +/- 2.8). Contrary to that observations in heifers, there was no increase in the number of 4- to 5-mm follicles in cows during superovulation. The number of larger follicles (>5 mm) increased during superovulation in both cattle groups, but it was significantly lower in cows than in heifers. During superovulation, the maximal E2 concentration was greater (P < 0.0001) in heifers than in cows. One cow showed delayed luteolysis during superovulation, while another had abnormally high FSH (>10 ng/ml) and LH (>3 ng/ml) concentrations following superovulation. All the cows had a postovulatory FSH rise which was not detected in the heifers. The results showed that attempts to improve superovulatory response in mature genetically valuable cows are hampered by a number of reproductive disorders that are not predictable from the study of the unstimulated cycle.

Journal Article↗

Induction of multiple follicular development as a therapy for unexplained or male-related infertility.

Most invasive techniques (i.e., in-vitro fertilization, gamete intrafallopian transfer) used for the therapy of unexplained or male-related infertility involve the induction of multiple follicular development (MFD). The efficacy of MFD in conjunction with a non-invasive technique (artificial insemination with husband's semen) was evaluated in a group of 41 couples with long-lasting infertility. MFD was obtained by means of clomiphene citrate and exogenous gonadotropins. Within 6 months of treatment, pregnancy was achieved by 52.2% of couples with male-related infertility and by 54.5% of couples with unexplained infertility. Sperm parameters, age of the female partner and preovulatory estradiol peak did not show any significant difference between couples who conceived and those who did not. It is concluded that induction of MFD represents an effective therapy and might replace invasive techniques for the treatment of the above conditions.

Adult↗

Follicular development during late human pregnancy.

In contrast to the accepted view of ovarian quiescence during pregnancy, the ovaries of preparturient women are covered with a dense population of small superficial follicles. In this study we have measured the follicular size and status of the oocytes and cumulus oophorus in 298 follicles from the ovaries of 30 women. The samples were taken at cesarean section and were examined with Nomarski differential interference microscopy. No oocyte was recovered from 50% of the follicles; 79% of the recovered oocytes and 78% of their cumuli were degenerative. Degeneration was correlated with appearance of phagocytes in the follicular fluid. These findings suggest that the endocrine status of gestation does not prevent early follicular development, but induces premature atresia.

Female↗

Previous cycle cancellation due to poor follicular development as a predictor of ovarian response in cycles stimulated with gonadotrophin-releasing hormone agonist-gonadotrophin treatment.

BACKGROUND: There is scanty information analysing the predictive value of a poor response, in terms of cancellation of the IVF cycle because of poor follicular development, as a predictor of ovarian response in a subsequent treatment cycle. This study, where logistic regression analysis was used, was undertaken to investigate the relative power of the woman's age, basal FSH, and previous cycle cancellation both as single and combined predictors of ovarian response in an IVF program where pituitary desensitization is routinely used. METHODS: One hundred and twenty-nine consecutive patients having their first cycle of IVF/ICSI treatment cancelled because of poor follicular response and undergoing a second attempt within 6 months after the failed treatment cycle were initially selected (group 1). Group 2 comprised 129 patients undergoing the first cycle of IVF/ICSI treatment and who were randomly selected from our assisted reproductive treatment program matching by BMI and indication for IVF/ICSI to those in group 1. RESULTS: Cancellation rate was significantly higher but ovarian response significantly lower in group 1 as compared with group 2. As indicated by the AUC(ROC) determined with ROC analysis, such a poor outcome in patients having a previous IVF/ICSI cycle cancelled due to poor response was observed whatever the level of basal FSH. In a logistic regression analysis and according to the odds ratio values, the predictive capacity of a previous poor response was 9 and 7.6 times higher than the predictive capacity of age and basal FSH, respectively. Any two or all three variables studied did not improve the predictive value of previous cycle cancellation alone. CONCLUSIONS: The history of an IVF/ICSI cancelled cycle due to poor follicular response in a standard stimulation protocol is a better predictor of cancellation in subsequent treatment cycles than age or FSH. The poor ovarian response associated with previous cycle cancellation occurs whatever the level of basal FSH.

Adult↗