Regulation of the follicular hierarchy and ovulation

Studies are discussed which investigate the regulation of follicular maturation and the ovulation sequence of the domestic hen. The number of FSH receptors of ovarian granulosa cells decreases as the follicle matures, and this decrease in receptor number is paralleled by a gradual loss of FSH-stimulable adenylyl cyclase (AC) activity. By contrast, LH-stimulable AC activity increases as the follicle progresses through the hierarchy. In addition, FSH stimulates progesterone secretion by granulosa cells of the smaller preovulatory follicles, whereas these cells are only minimally responsive to LH. These data suggest that the maturation of less mature (smaller) follicles is primarily controlled by FSH, while LH may serve primarily as the ovulation-inducing hormone. The ability of LH to stimulate progesterone release and induce premature ovulation is dependent upon the stage of the sequence. Injection of ovine LH 12 hr prior to ovulation of the first (C1) egg of the sequence induces fully potentiated preovulatory plasma progesterone surges and 100% premature ovulation, whereas injection prior to the second (C2) ovulation of the sequence fails to stimulate prolonged progesterone release and induces premature ovulation in less than 50% of injected hens. These results are consistent with data obtained in vitro which suggest that granulosa cells obtained 12 hr prior to a C1 ovulation secrete more progesterone in response to chicken LH compared to those obtained 12 hr prior to the C2 ovulation. These data are discussed in terms of the ovary's ability to act as a regulator of the ovulatory cycle.     

Effects of follicular status at treatment on follicular development and ovulation in response to FSH in Spanish merino ewes

Cyclic Spanish Merino ewes were treated on Day 13 of the estrous cycle with 12 mg, i.m., FSH-P in saline (n = 9) or propylene glycol (n = 24), currently with 100 micrograms, i.m., Cloprostenol (Day 0). From Day-6 to Day 0, the ewes were observed daily by transrectal ultrasonography, after Day 0, ultrasonography was performed every 12 h for 72 h. Sizes and locations of > or = 2 mm follicles were recorded at each observation. The ovulation rate was determined by laparoscopy on Day 7 after estrus. The number of ovulations ranged from 0 to 6 in ewes treated with FSH-P in saline and from 0 to 16 in ewes receiving FSH-P in propylene glycol (P < 0.05). In the latter group, the response was bimodally distributed; about half of the females had 1 ovulation, whereas the remainder had > 4 with a mean of 7 ovulations. The ovulation rate was associated with 2 characteristics of the largest follicle present at treatment (Day 0). First, if the largest follicle on Day 0 had not changed in diameter from Day-1 to Day 0, then 7 of 9 ewes had > 3 ovulations; if the largest follicle had either increased or decreased, only 8 of 24 ewes had > 3 ovulations (P < 0.05). Second, there was a linear trend (P < 0.07) for ovulation rate to decrease as the persistence of the largest follicle at treatment increased; no ewe in which the largest follicle on Day 0 remained present for more than 36 h ovulated more than 6 follicles. As with the ovulation rate, the numbers of large follicles on Days 1.5, 2 and 2.5 varied with the interaction of change in diameter of the largest follicle on Day 0 from Day-1 to Day 0 and with vehicle. In summary, the superovulatory response was affected by the change in diameter from Day-1 to Day 0 of the largest follicle on Day 0 and the period required for that follicle to regress after treatment with FSH-P and cloprostenol.     

Structure and function of the amphibian follicular epithelium during ovulation

Summary Low concentrations of cytochalasin B (CCB) are known to inhibit ovulation in the frog, Hyla regilla. Examination of amphibian thecal cell ultrastructure reveals filaments (average diameter 71 Å) arranged in bundles parallel to the surface of the oocyte. These filaments are often associated with hemidesmosome-like plaques on the basal plasmalemma. While individual filaments appear unaltered morphologically by CCB (1–5 g/ml), their organization into bundles, apparent relationship to the hemidesmosomes, and the highly contorted configuration of the thecal cells after oocyte expulsion, suggest that a nonmuscular contractile system residing within the follicle plays a fundamental role in ovulation.Our data suggest that the flattened epithelioid thecal cells shorten all axes that run parallel to the oocyte surface via filament bundle contractions, while they remain tightly bound together by macular attachment plaques. These cells thus increase in height to become cuboidal-low columnar in shape; the area covered by the base of each is greatly reduced. As this thecal sac decreases in size, the compression generated by the contractile mechanism forces the oocyte through the enzymatically weakened apex of the follicle and ovulation results.This investigation was supported by grant HD-07194 from the National Institutes of Health. The authors are grateful to Dr. Arthur L. Cohen, Director of the Electron Microscope Center for use of the Center's scanning electron microscope. We are also indebted to Mrs. Gail M. McDole and Mr. James D. Huber for able technical assistance     

Use of bovine follicular fluid to increase ovulation rate or prevent ovulation in sheep

Romney ewes were injected intramuscularly once or twice daily for 3 days with 0, 0.1, 0.5, 1 or 5 ml of bovine follicular fluid (bFF) treated with dextran-coated charcoal, starting immediately after injection of cloprostenol to initiate luteolysis on Day 10 of the oestrous cycle. There was a dose-related suppression of plasma concentrations of FSH, but not LH, during the treatment period. On stopping the bFF treatment, plasma FSH concentrations 'rebounded' to levels up to 3-fold higher than pretreatment values. The mean time to the onset of oestrus was also increased in a dose-related manner by up to 11 days. The mean ovulation rates of ewes receiving 1.0 ml bFF twice daily (1.9 +/- 0.2 ovulations/ewe, mean +/- s.e.m. for N = 34) or 5.0 ml once daily (2.0 +/- 0.2 ovulations/ewe, N = 25) were significantly higher than that of control ewes (1.4 +/- 0.1 ovulations/ewe, N = 35). Comparison of the ovaries of ewes treated with bFF for 24 or 48 h with the ovaries of control ewes revealed no differences in the number or size distribution of antral follicles. However, the large follicles (greater than or equal to 5 mm diam.) of bFF-treated ewes had lower concentrations of oestradiol-17 beta in follicular fluid, contained fewer granulosa cells and the granulosa cells had a reduced capacity to aromatize testosterone to oestradiol-17 beta and produce cyclic AMP when challenged with FSH or LH. No significant effects of bFF treatment were observed in small (1-2.5 mm diam.) or medium (3-4.5 mm diam.) sized follicles. Ewes receiving 5 ml bFF once daily for 27 days, from the onset of luteolysis, were rendered infertile during this treatment period. Oestrus was not observed and ovulation did not occur. Median concentrations of plasma FSH fell to 20% of pretreatment values within 2 days. Thereafter they gradually rose over the next 8 days to reach 60% of pretreatment values where they remained for the rest of the 27-day treatment period. Median concentrations of plasma LH increased during the treatment period to levels up to 6-fold higher than pretreatment values. When bFF treatment was stopped, plasma concentrations of FSH and LH quickly returned to control levels, and oestrus was observed within 2 weeks. The ewes were mated at this first oestrus and each subsequently delivered a single lamb.     

A model of follicular development and ovulation in sheep and cattle

A dynamic model to describe ovarian follicular development following commitment has been developed. It identifies follicular growth with oestradiol production and assumes that this growth is the result of intra-ovarian stimulation, gonadotrophin stimulation, and inhibitory interactions among the follicles, where larger follicles suppress the growth of the smaller follicles. The variables of the model are the levels of oestradiol in each follicle at commitment, the rate of change of oestradiol production by individual follicles during follicular development, and the level of oestradiol that will induce luteinizing hormone (LH) surge. Changes in the variables of the model could be associated with both genetic and environmental effects. The behaviour of the model is consistent with experimental observations. The model can be expanded to include exogenous follicle-stimulating hormone (FSH) administration assuming that FSH is associated with advancing the maturation of gonadotrophin-dependent follicles without affecting the number of committed follicles. The use of the model to explore FSH administration strategies is demonstrated. The model confirms that the response to FSH administration depends on both the amount of FSH and the time of administration. The largest number of double ovulations occurred when FSH was given at the time of the deviation of the dominant and subordinate follicles.     

Delayed follicular development and ovulation following inhibition of FSH with equine follicular fluid in the mare

In two experiments, PGF(2alpha) was given to all mares on Day 10 (ovulation = Day 0). In experiment 1, mares received either whole follicular fluid or saline i.v. every 12 hours on Days 10 to 14. Experiment 2 was similar to experiment 1, except the follicular fluid was extracted with charcoal to remove steroids. Analysis of the FSH data for Days 10 to 21 indicated an effect of treatment (P<0.08) with whole follicular fluid, but not with charcoal-extracted follicular fluid. However, there was an effect of day (P<0.05) and an interaction (P<0.01) of treatment with day for both experiments. The interaction of treatment with day seemed primarily due to a marked post-treatment increase in FSH concentrations between Days 15 and 17 for mares treated with either whole follicular fluid or charcoal-extracted follicular fluid. Analysis of the diameter of the largest follicle for Days 10 to 18 indicated a main effect of treatment (P<0.05) and day (P<0.05) and an interaction (P<0.05) of treatment with day for both experiments. The interaction of treatment with day was attributable to the inhibition of follicular growth by Day 14 for mares treated with whole follicular fluid and by Day 15 for mares treated with charcoal-extracted follicular fluid. The length of the interovulatory interval was longer (P<0.05) in the treated group than in controls for both experiments. Results indicated that equine follicular fluid contained a proteinaceous substance that suppressed circulating concentration of FSH. The inhibited follicular growth and the delay in ovulation were attributed to the reduced concentrations of circulating FSH.     

The morphology of follicular development and ovulation in non-human primates.

A colony of Macaca fascicularis have been under continuous observation for the past 4 years to ascertain the follicular morphological changes that occur prior to ovulation and during the development of the corpus luteum. Of 609 experimental cycles, laparoscopy was performed at least once in 44-5% of the cycles. Of the 104 cycles where the ovulatory status was definitely known, 89-4% were deemed ovulatory and 10-6% anovulatory. The presence or absence of ovulation in the previous cycle did not have an effect on the cycle length either for the total cycles or when analysing only cycles over 28 days. Similarly, the occurrence of two consecutive ovulations (in consecutive cycles) on the same vs. opposite ovaries did not have a significant effect on the cycle length. Neither laparoscopic stress or anaesthesia effected the normal cyclicity of the animals. The characteristic changes in follicular morphology are most clearly defined in M. fascicularis. In this species the 24 of 36 hr prior to ovulation are accompanied by discrete changes which occur in a fixed sequence, allowing one to predict the time of ovulation with reasonable accuracy. In S. sciureus ovulation is preceded by extensive bulging at the follicular apex and haemorrhaging at the base of the follicle. Due to this haemorrhaging post-ovulatory follicles in S. sciureus are generally more easily discernible than in fascicularis. The formation of clear areas (stigma) is not as evident in either S. sciureus or G. senegalensis as in the macaque. Actual ovulation has been observed four times in M. fascicularis and twice in S. sciureus.     

A model for follicular selection and ovulation: lessons from superovulation

A model for selection of the preovulatory follicle during the normal ovarian cycle is proposed. During menstruation the concentration of FSH rises to a level high enough to "activate" a single small antral follicle (2-4 mm dia.) so that it can produce large amounts of oestradiol. As the follicle develops, the concentration of FSH is suppressed below this threshold level by the secretion of oestradiol and inhibin. The dominant follicle becomes increasingly sensitive to FSH so that it continues to develop in an environment which inhibits development of other follicles. Multiple ovulation can be achieved by extending the period during which the level of FSH remains above this threshold level (e.g. during treatment with clomiphene or gonadotrophins). Although multiple ovulation occurs when the gate is widened in this way, the follicles are never completely synchronous as they continue to grow at approximately the same rate. Current evidence suggests that ovulation occurs at random between the two ovaries in successive cycles and that the corpus luteum exerts an inhibitory effect on folliculogenesis by suppressing the secretion of FSH and LH. These observations are compatible with the hypothesis that while small antral follicles are recruited continuously, at all stages of reproductive life, selection of the dominant follicle requires the unique gonadotrophic environment which is only present in the early follicular phase. The follicle of the month is, therefore, selected by chance because it is at the right place at the right time.     

Regulation of follicular activity and ovulation in ewes by exogenous progestagen

The success of estrus synchronization programs using progestagen sponges, particularly for fixed-time AI, varies considerably. In view of the recent evidence in cattle that exogenous progestins alter follicular dynamics, it may be that the stage of the estrous cycle at which the synchronization protocol is begun affects the synchrony of ovulation. The goal of this study was to evaluate the effect of medroxyprogesterone acetate (MAP) intravaginal sponges on follicular dynamics, luteal function and interval to ovulation when inserted at 3 stages of the estrous cycle. Sponges were inserted for 12 d beginning on either Day 0, 6 or 12 (n = 5) following ovulation. Ovarian activity was monitored using real-time ultrasound imaging during the treatment and the post-treatment estrous cycles. Information from the post-treatment cycle was used as a baseline to compare with the treatment cycle. Most ewes (79%) in the post-treatment cycle exhibited 3 follicular waves in an estrous cycle of 16 d, with the second wave follicles having smaller diameter (P < 0.001). Treatment with MAP increased the number of follicular waves from 3 to 4 or 5 when sponges were inserted on Days 6 and 12, respectively. Size of the largest follicle was smaller (P > 0.01) in waves in the early and middle of the 12-d MAP treatment period when compared with the last 4 days. This effect was most pronounced when endogenous progesterone concentrations were elevated concurrently with the presence of the sponge. Persistence of the ovulatory follicle was increased (P < 0.001) when sponges were inserted on Day 12, the only treatment where these follicles were under the influence of MAP in the absence of functional corpora lutea. Follicles were regressing at sponge removal in the Day 6 treatment, which resulted in a delay in emergence of ovulatory follicles, the LH surge and ovulation (P < 0.08) in relation to Day 0 and Day 12. Treatment with MAP sponges does not adequately synchronize estrus and ovulation among cyclic ewes due to the different follicular patterns that result depending on the stage of cycle at the time of sponge insertion.     

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.     

Total follicular populations in ewes of high and low ovulation rates.

The total ovarian follicular populations were studied in two breeds of ewes which differed greatly in their ovulation rates. Thus 8 Romanov (mean ovulation rate 3.1) and 12 Ile-de-France ewes (mean ovulation rate 1.4) were ovariectomized at oestrus during the breeding season. Each right ovary and 3 left ovaries were sectioned at 7 micron and examined microscopically. The number of small follicles, i.e. with 2 or less layers of granulosa cells, was estimated by a tested sampling procedure whilst all larger follicles were measured and arranged into classes. There were half as many small follicles but 1.5--2 times more large follicles in the ovaries of the Romanov ewes compared to those of the Ile-de-France ewes. The number of atretic follicles was approximately the same in both breeds and does not explain the difference observed in ovulation rate. It is concluded that the higher ovulation rate in the Romanov ewe is due to the greater number of large follicles available to be stimulated for ovulation.     

Gonadotropin-releasing hormone treatment induces follicular growth and ovulation in seasonally anestrous mares

A study was conducted to evaluate the effectiveness of gonadotropin-releasing hormone (GnRH) pulse infusion to stimulate follicular development and induce ovulation in seasonally anestrous standardbred mares. Seventeen mares were selected for use in this experiment, on the basis of a previous normal reproductive history, and were housed under a photoperiod of 8L:16D beginning one week prior to the start of the experiment (second week in January). Mares were infused with 20 micrograms (n = 7) or 2 micrograms (n = 6) GnRH/h, or were subjected to photoperiod treatment only (controls, n = 4). Serum concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH), and progesterone did not vary, and neither significant follicular development nor ovulation was observed in any control mare throughout the experimental period (greater than 60 days). By contrast, both groups of GnRH-treated mares showed elevated serum concentrations of LH and FSH within one day after the start of infusion. Mares infused with 20 micrograms GnRH/h had at least one follicle greater than or equal to 25 mm in 7.4 +/- 1.3 (mean +/- SEM) days following the start of infusion, and ovulated in 12.0 +/- 0.7 days. In the 2-microgram-GnRH/h treatment group, a 25-mm follicle was detected in 5.7 +/- 0.7 days, and ovulation occurred after 10.0 +/- 0.3 days of infusion. Ovulation in every instance was followed by a functional luteal phase, as indicated by the profiles of progesterone secretion.(ABSTRACT TRUNCATED AT 250 WORDS)     

Ovarian follicular wave synchronization and induction of ovulation in postpartum beef cows

An experiment was designed to evaluate a) the effect of a progesterone-estradiol combined treatment on ovarian follicular dynamics in postpartum beef cows, and b) ovulation and the subsequent luteal activity after short-term calf removal and GnRH agonist treatment. Multiparous Angus cows (25 to 40 d after calving) were assigned to the following treatments: untreated (Control, n = 9); short term calf removal (CR, n = 8); progesterone (CIDR, n = 9) and progesterone plus estradiol-17 beta (CIDR + E-17 beta, n = 9). Progesterone treatment (CIDR) lasted 8 d and the day of device insertion was considered as Day 0. Cows in the CIDR + E-17 beta group also received an i.m. injection of 5 mg of E-17 beta on Day 1. On Day 8, calves were removed for 48 h (CR, CIDR and CIDR + E-17 beta groups) and 6 h before the end of calf removal these cows also received an i.m. injection of 8 micrograms of Busereline (GnRH). Anestrus was confirmed in all cows by the absence of luteal tissue and progesterone concentrations below 1 ng ml-1 at the beginning of the experiment. Although mean (+/- SEM) interval from the beginning of the experiment (Day 0) to wave emergence did not differ (P > 0.05) among treatment groups (Control, 1.9 +/- 1.0, range -2 to 7 d; CR, 3.9 +/- 0.7, range 0 to 6 d; CIDR, 2.8 +/- 0.5, range 0 to 4 d and CIDR + E-17 beta, 4.1 +/- 0.2, range 3 to 5), the variability was less (P < 0.05) in the CIDR + E-17 beta group. The proportion of cows ovulating 24 to 48 h after GnRH administration tended (P = 0.08) to be higher in cows from CIDR + E-17 beta group (8/9) than in those of CR (5/8) or CIDR (6/9) groups, respectively and was associated with a higher proportion (P < 0.05) of CIDR + E-17 beta treated cows (9/9) that had a dominant follicle in the growing/early static phase at the time of GnRH treatment compared to the other GnRH treated groups (5/8, and 4/9 for CR and CIDR groups, respectively). Two CR cows ovulated 0-24 h after GnRH and only one Control cow ovulated the day before the time of GnRH administration. Cows pretreated with progesterone had longer (P < 0.05) luteal lifespan (CIDR, 14.5 +/- 0.7, CIDR + E-17 beta, 13.9 +/- 0.6 d) than those not treated with CIDR (Control, 5, CR, 4.0 +/- 0.4). We conclude that progesterone plus estradiol treatment results in tightly synchronized wave emergence and high GnRH-induced ovulation rate with normal luteal activity in postpartum beef cattle.     

Lacker's model: control of follicular growth and ovulation in domestic species

Lacker (1981) and Lacker & Akin (1988) developed a mathematical model of follicular maturation and ovulation; this model of only four parameters accounts for a large number of results obtained over the past decade or more on the control of follicular growth and ovulation in mammals. It establishes a single law of maturation for each follicle which describes the interactions between growing follicles. The function put forward is sufficient to explain the constancy of the number of ovulations or large follicles in a female as well as the variability of this number among strains or species and for either induced or spontaneous ovulators. According to the model, the number of ovulations or large follicles lies between two limits that are themselves simple functions of two parameters of the model. Moreover, Lacker's model exhibits interesting characteristics in agreement with results obtained by physiologists: in particular, it predicts that the number of ovulating or large follicles is independent of:

1. the total number of maturing follicles,
2. the process of recruitment of newly maturing follicles towards the terminal maturation (Poisson or other),
3. the form of the LH or FSH secretion curves as functions of the systemic level of oestradiol. The model further predicts that
4. selection and dominance of follicles result from the feedback between the ovary and the hypophysis through the interactions between follicles; these interactions are expressed by the maturation function of the model.
5. recovery from atresia is possible for a follicle: from decreasing, the rate of secretion of oestradiol may increase.
6. the revised model suggests a renewal of follicles during the sexual cycle, as “waves of follicular growth”.

Lacker's model is a model of strict dominance; it maintains a hierarchy of the follicles as soon as they start their final maturation to the ovulations as that is observed in bird or reptile ovary. Such a strict hierarchy is possible but it is probably not a general situation in all mammals. We therefore modified the maturing function of the follicle in order to make it compatible with the observations of physiologists: follicles always interact as in the initial model but they individually become old, the hierarchy of follicles can be modified with time and the largest follicles do not indefinitely grow as in the initial model.