Regulation of ovarian follicular dynamics in farm animals. Implications for manipulation of reproduction

In this review, the main features of folliculogenesis are summarized and compared among species. In the past few years, ultrasonography has clarified follicle growth patterns, and our understanding of follicle maturation has improved considerably. As the follicles develop towards the ovulatory stage, three features appear to be highly conserved across all species: 1) the sequence of events (recruitment, selection and dominance); 2) the sequential need for gonadotropins (FSH for recruitment, LH for dominance) and 3) the large variability of numerical parameters (number of waves per cycle, number of follicles per wave) as well as temporal requirements (time of selection, duration of dominance). In addition, specific follicles may also have variable gonadotropin requirements (thresholds). When patterns of follicle development at different physiological states are compared across species, follicular waves were detected in cattle, sheep and horses and during the prepubertal period in swine, suggesting that ovaries of all species operate on a wave basis unless they are prevented from doing so. Efficient estrus control treatments should have the ability to affect 1) the wave pattern by preventing the development of persistent dominant follicles containing aging oocytes, and 2) the recruitment of the future ovulatory follicle whatever the stage of the wave at the time of treatment. This would allow synchronous ovulation of a growing dominant follicle. Manipulation of the luteal phase follicular waves after mating or AI may also optimize fertility. Superovulation is still an efficient technique to obtain progeny from genetically valuable females. Administration of exogenous gonadotropins acts to reveal the underlying ovarian variability. Ovarian response of each female depends on the number of gonado-sensitive follicles present at the time when treatment is initiated. Identification of the number of such follicles for each female would improve efficacy of superovulation, by allocating potential nonresponders to other techniques (OPU/FIV). One of the main components of the within female response to superovulation is the stage of the wave when gonadotropins are injected. Treatment in the absence of a dominant follicle ensures a response close to the female's specific maximum. The development of practical approaches to achieve this still requires further research.     

Short- and long-term effects of unilateral ovariectomy in sheep: causative mechanisms

The mechanisms of ovulatory compensation following unilateral ovariectomy (ULO) are still not understood. In the present study, we investigated the short- and long-term effects of ULO in sheep using transrectal ovarian ultrasonography and hormone estimations made during the estrous cycle in which surgery was done, the estrous cycle 2 mo after surgery, and the 17-day period during the subsequent anestrus. The ULOs were done when a follicle in the first follicular wave of the cycle reached a diameter > or =5 mm, leaving at least one corpus luteum and one ovulatory-sized follicle in the remaining ovary. Ovulation rate per ewe was 50% higher in the ULO ewes compared with the control ewes at the end of the cycle during which surgery was performed, but it did not differ between groups at the end of the cycle, 2 mo later. This compensation of ovulation rate in ULO ewes was due to ovulation of follicles from the penultimate follicular wave in addition to those from the final wave of the cycle. Ovulation from multiple follicular waves appeared to be due to a prolongation of the static phase of the largest follicle of the penultimate wave of the cycle. Interestingly, the length of the static phase of waves was prolonged in ULO ewes compared with control ewes in every instance where the length of the static phase could be determined. Changes in follicular dynamics due to ULO were not associated with alterations in FSH and LH secretion. In conclusion, ovulatory compensation in ULO sheep involves ovulation from multiple follicular waves due to the lengthened static phase of ovulatory-sized follicles. These altered antral follicular dynamics do not appear to be FSH or LH dependent. Further studies are required to examine the potential role of the nervous system in the enhancement of the life span of the ovulatory-sized follicles leading to ovulatory compensation by the unpaired ovary in ULO sheep.     

The temporal relationship between patterns of LH and FSH secretion, and development of ovulatory-sized follicles during the mid- to late-luteal phase of sheep

The aim of the present study was to investigate the temporal relationship between the secretory pattern of serum LH and FSH concentrations and waves of ovarian antral follicles during the luteal phase of the estrous cycle in sheep. The growth pattern of ovarian antral follicles and CL were monitored by transrectal ultrasonography and gonadotropin concentrations were measured in blood samples collected every 12 min for 6 h/d from 7 to 14 d after ovulation. There were two follicular waves (penultimate and final waves of the cycle) emerging and growing during the period of intensive blood sampling. Mean and basal LH concentrations and LH pulse frequency increased (P < 0.001) with decreasing progesterone concentration at the end of the cycle. Mean and basal FSH concentrations reached a peak (P < 0.01) on the day of follicular wave emergence before declining to a nadir by 2 d after emergence. None of the parameters of pulsatile LH secretion varied significantly with either the emergence of the final follicular wave or with the end of the growth phase of the largest follicle of the penultimate wave of the cycle. However, mean and basal LH concentrations did increase (P < 0.05) after the end of the growth phase of the largest follicle of the final follicular wave of the cycle. Furthermore, the end of the growth phase of the largest follicle of the final wave coincided with functional luteolysis. In summary, there was no abrupt or short-term change in pulsatile LH secretion in association with the emergence or growth of the largest follicle of a wave. We concluded that the emergence and growth of ovarian antral follicles in follicular waves do not require changes in LH secretion, but may involve changes in sensitivity of ovarian follicles to serum LH concentrations.     

Ovarian follicular dynamics during the estrous cycle in heifers monitored by real-time ultrasonography

It is not clear whether the turnover of ovarian follicles during the estrous cycle in cattle is continuous and independent of the phase of the cycle, or whether waves of follicular growth occur at specific times of the cycle. To clarify this controversy, the pattern of growth and regression of ovarian follicles was characterized during a complete estrous cycle in ten heifers by daily ultrasonographic examinations. Follicles greater than or equal to 5 mm were measured and their relative locations within the ovary were determined in order to follow the sequential development of each individual follicle. Results indicated the presence of either two (n = 2 heifers), three (n = 7), or four (n = 1) waves of follicular growth per cycle. Each wave was characterized by the development of one large (dominant) follicle and a variable number of smaller (non-dominant) follicles. In the most common pattern observed (three waves/cycle), the first, second, and third waves started on Days 1.9 +/- 0.3, 9.4 +/- 0.5, and 16.1 +/- 0.7 (X +/- SEM), respectively. The dominant follicle in the third wave was the ovulatory follicle. The maximal size and the growth rate of the dominant follicle in the second wave were significantly lower than in the other waves, but no significant difference was observed between the first and third waves. For the two heifers that had two follicular waves/cycle, the waves started on Days 2 and 11, whereas in the remaining heifer (four waves/cycle), the waves began on Days 2, 8, 14, and 17, respectively. At 0, 1, 2, 3, and 4 days before estrus, the ovulatory follicle was the largest follicle in the ovaries in 100%, 95%, 74%, 35%, and 25% of follicular phases monitored, respectively. The relative size of the preovulatory follicle at the completion of luteolysis (progesterone less than 1 ng/ml) was negatively correlated (r = -0.90; p less than 0.0001) with the interval of time between the end of luteolysis and the luteinizing hormone surge, suggesting that the length of proestrus is determined by the size of the pre-ovulatory follicle at the beginning of proestrus. In conclusion, this study shows that the development of ovarian follicles greater than or equal to 5 mm in heifers occurs in waves and that the most common pattern is three waves per estrous cycle.     

Pattern of follicular development in high fecundity Olkuska ewes during the estrous cycle

Folliculogenesis was studied daily in the 18 oestrous cycles in six prolific Olkuska ewes from October to December using transrectal ultrasonography to record the number and size of all ovarian follicles > or =2 mm in diameter. Blood samples were taken once a day and were analyzed for concentrations of FSH, LH, estradiol and progesterone. Follicular and hormonal data were analyzed for associations between different stages of development of the follicular waves and concentrations of FSH and estradiol. The first wave during which at least one follicle reached maximum diameter of > or =4 mm after ovulation, was defined as a wave 1, and the following waves were numbered sequentially. Waves 1, 2, 3, 4 and the ovulatory one emerged on days: -2 to 4, 4 to 8, 6 to 11, 10 to 12 and 11 to 15, respectively. The mean number of follicles per wave that reached diameter of > or =4 mm was 4.15 +/- 1.1 and 16.62 +/- 8.6 follicles per estrous cycle of a total 299 follicles were observed. Significantly more follicles (p> or =0.05) emerged on days 2, 8 and 13 than in other days. Serum FSH concentrations fluctuated from 0.11 ngml(-1) on day 2 to preovulatory maximum 1.81 ngml(-1) on day 17 of the estrous cycle. The emergence of follicular waves was associated with elevations of FSH concentrations in blood serum. The mean increase in FSH concentration was followed by the recruitment of follicles of the next wave. The mean daily FSH concentration and the mean number of follicles emerging each day were negatively correlated. The length of the interwave interval (4.4 +/- 1.6 days) did not differ significantly from the interval between pulses of FSH (4.8 +/- 0.3 days). The mean serum estradiol concentrations showed fluctuations until day 14 and then gradually increased from 5.47 +/- 0.3 pgml(-1) to reach a peak 13.14 +/- 0.2 pgml(-1) on the day before ovulation. To summarize, the growth of ovarian follicles during the estrous cycle in high fecundity Olkuska sheep exhibited a distinct wave-like pattern. Ovarian follicles emerged from the pool of 2 mm follicles. The preovulatory follicles originated from the large follicle population were present in the ovary at the time of luteal regression. The initial stages of the growth of the largest follicles appears to be controlled primarily by increases in FSH secretion.     

Characterization of ovarian follicular wave dynamics in women

A wave phenomenon of ovarian follicular development in women has recently been documented in our laboratory. The objective of the present study was to characterize follicular waves to determine whether women exhibit major and minor wave patterns of follicle development during the interovulatory interval (IOI). The ovaries of 50 women with clinically normal menstrual cycles were examined daily using transvaginal ultrasonography for one IOI. Profiles of the diameters of all follicles >or=4 mm and the numbers of follicles >or=5 mm were graphed during the IOI. Major waves were defined as those in which one follicle grew to >or=10 mm and exceeded all other follicles by >or=2 mm. Minor waves were defined as those in which follicles developed to a diameter of <10 mm and follicle dominance was not manifest. Blood samples were drawn to measure serum concentrations of estradiol-17beta, LH, and FSH. Women exhibited major and minor patterns of follicular wave dynamics during the IOI. Of the 50 women evaluated, 29/34 women with two follicle waves (85.3%) exhibited a minor-major wave pattern of follicle development and 5 women (14.7%) exhibited a major-major wave pattern. Ten of the 16 women with three follicle waves (62.5%) exhibited a minor-minor-major wave pattern, 3 women (18.8%) exhibited a minor-major-major wave pattern, and 3 women (18.8%) exhibited a major-major-major wave pattern. Documentation of major and minor follicular waves during the menstrual cycle challenges the traditional theory that a single cohort of antral follicles grows only during the follicular phase of the menstrual cycle.     

The ability of subordinate follicles of the second follicular wave to become dominant is lost by day 15 of the estrous cycle in cattle

Generally, unilateral ovariectomy before a critical period in the latter part of the estrous cycle induces a transitory increase in plasma FSH, which causes subordinate follicles to develop and maintain ovulation rates characteristic of the species. A limiting period for subordinate follicles to assume dominance and from which ovulation occurs has not been shown for cattle. Growth and/or regression of subordinate follicles were characterized following removal of the dominant follicle at different days of the luteal phase of the estrous cycle in cattle in this study. In the mid-luteal phase (Day 13 or 15), the ovary with the dominant follicle of the second wave was ablated via unilateral ovariectomy; the corpus luteum also was removed. In the late luteal phase (Day 17 or 19), the dominant follicle was ablated with an ultrasonically guided 20 gauge needle. When the dominant follicle was removed on Day 13, the largest subordinate follicle of the second wave of follicular development became dominant and ovulation occurred from this follicle in 4 of 4 animals. However, when the dominant follicle was removed on Day 15, 17 or 19, a new wave of follicular development was induced in 14 of 15 animals. Moreover, the recovered subordinate follicle of the second wave of follicular development had similar growth characteristics to naturally occurring dominant follicles. In conclusion, the subordinate follicle in the second follicular wave in cattle retained the ability to become dominant, but this ability was lost by Day 15 of the estrous cycle. However, cattle then were able to maintain ovulation by developing a new wave of follicular growth.     

Reproductive cycles in sheep

During the last three decades, there has been remarkable progress in many aspects of ovarian biology due to advances in real-time ultrasonography, which permits non-invasive, repeated monitoring of ovarian structures in conscious and non-anaesthetised animals. This review is primarily concerned with ovarian activity, as determined by transrectal ultrasonography, and measurements of circulating concentrations of gonadotrophins and ovarian steroids during reproductive cycles in sheep. The growth of antral follicles reaching ostensibly ovulatory sizes occurs in a wave-like pattern throughout the breeding season in both prolific and non-prolific breeds of sheep. There are typically 3 or 4 waves of follicle development during the interovulatory interval. Follicular wave emergence is primarily controlled by changes in circulating concentrations of follicle-stimulating hormone (FSH) but diminished ovarian responsiveness to gonadotrophic signals may result in reduced numbers of follicular waves. In cyclic ewes, the largest ovarian follicles acquire the ability to secrete oestradiol from the day of emergence with peak oestradiol secretion occurring about the time they reach maximum diameter. The high ovulation rate in some prolific breeds may be achieved by the ovulation of follicles from the last two waves of the interovulatory interval. Prolific ewes tend to produce more but smaller corpora lutea (CL) and have lower serum concentrations of progesterone during the luteal phase of the oestrous cycle as compared to less prolific genotypes. Lastly, recent studies of the endocrine influences on ovarian function have brought into question the existence of strong follicular dominance, as seen in cattle, and provided new insights into the effects of luteal progesterone on antral follicular development in ewes.     

An ultrasound-aided study of temporal relationships between the patterns of LH/FSH secretion, development of ovulatory-sized antral follicles and formation of corpora lutea in ewes

To characterize the pulsatile secretion of LH and FSH and their relationships with various stages of follicular wave development (follicles growing from 3 to > or =5 mm) and formation of corpora lutea (CL), 6 Western white-faced ewes underwent ovarian ultrasonography and intensive blood sampling (every 12 min for 6 h) each day, for 10 and 8 consecutive days, commencing 1 and 2 d after estrus, respectively. Basal serum concentrations of LH and LH pulse frequency declined, whereas LH pulse duration and FSH pulse frequency increased by Day 7 after ovulation (P<0.05). LH pulse amplitude increased (P<0.05) at the end of the growth phase of the largest ovarian follicles in the first follicular wave of the cycle. The amplitude and duration of LH pulses rose (P<0.05) 1 d after CL detection. Mean and basal serum FSH concentrations increased (P<0.05) on the day of emergence of the second follicular wave, and also at the beginning of the static phase of the largest ovarian follicles in the first follicular wave of the cycle. FSH pulse frequency increased (P<0.05) during the growth phase of emergent follicles in the second follicle wave. The detection of CL was associated with a transient decrease in mean and basal serum concentrations of FSH (P<0.05), and it was followed by a transient decline in FSH pulse frequency (P<0.05). These results indicate that LH secretion during the luteal phase of the sheep estrous cycle reflects primarily the stage of development of the CL, and only a rise in LH pulse amplitude may be linked to the end of the growth phase of the largest follicles of waves. Increases in mean and basal serum concentrations of FSH are tightly coupled with the days of follicular wave emergence, and they also coincide with the end of the growth phase of the largest follicles in a previous wave, but FSH pulse frequency increases during the follicle growth phase, especially at mid-cycle.     

Plasma concentrations of inhibin a and follicle-stimulating hormone differ between cows with two or three waves of ovarian follicular development in a single estrous cycle

Patterns of ovarian follicle development were monitored daily in Holstein-Friesian cows that had two (n = 4) or three (n = 4) waves of ovarian follicle development during a single estrous cycle. The plasma from daily blood samples was used in assays for inhibin A, FSH, progesterone, and estradiol-17beta. Mean cycle lengths for cows with two and three waves were 21.8 and 25.3 days, respectively (P < 0.02). Although the average number of follicles >3-mm diameter on each pair of ovaries was similar for two- and three-wave cows on Days 2, 3, and 4 (Day 0 = day of ovulation; 8.6 vs. 9.6 follicles), there were more follicles >6-mm diameter on the ovaries of cows with two waves on Days 3 and 4. This difference was associated with a shorter interval from wave emergence to peak concentrations of inhibin A during the first wave in two-wave cows (2.0 vs. 3.8 days; P = 0.03) and with higher peak concentrations (474 vs. 332 pg/ml; P = 0.03). Differences in peak FSH concentrations were not significant (1.7 vs. 1.3 ng/ml; P = 0.10) and were inversely related to inhibin A concentrations. The peak concentrations of inhibin A and FSH in the second nonovulatory wave in the three-wave cows were similar to the low concentrations measured in the first wave (292 vs. 332 pg/ml of inhibin A, 1.3 vs. 1.3 ng/ml of FSH; P > 0.20). Average peak concentrations of inhibin A and FSH were similar during the ovulatory wave for cows with either two or three waves in a cycle (432 vs. 464 pg/ml of inhibin A, 2.3 vs. 2.1 ng/ml of FSH; P > 0.3). The lower concentrations of FSH during the emergence of the first follicular wave in cows with three-wave cycles may have reduced the rate of development of some of the follicles and reduced the concentrations of inhibin A. This pattern of lower concentrations of FSH and inhibin A was repeated in the second nonovulatory wave but not in the ovulatory wave. Subtle differences in the concentrations of these two hormones may underlie the mechanism that influences the number of waves of ovarian follicle development that occur during the bovine estrous cycle.     

Ovarian follicular dynamics during the estrous cycle in buffalo (Bubalus bubalis)

The growth, selection, regression and ovulation of ovarian follicles was ultrasonically monitored in 30 Murrah buffalo throughout a spontaneous estrous cycle during the breeding season (autumn). Examinations revealed that follicular growth during the estrous cycle occurs in waves; the buffalo showed 1-wave (3.3%, n = 1), 2-wave (63.3%, n = 19) or 3-wave (33.3%, n = 10) follicular growth. The first wave began at 1.00, 1.16 +/-0.50 and 1.10 +/- 0.32 d in buffalo with 1, 2 and 3 waves, respectively (ovulation = Day 0). The second wave appeared at 10.83 +/- 1.09 and 9.30 +/- 1.25 d (P < 0.01) for the 2 and 3 wave cycle animals, respectively. The third wave started at 16.80 +/- 1.22 d. Structural persistence of the first dominant follicle was longer in the 2- than 3-wave cycles (20.67 +/- 1.18 vs 17.90 +/- 3.47 d ; P < 0.05). The duration of the growth and static phases of the first dominant follicle differed between the 2 and 3 wave cycles (P < 0.05), whereas there were no differences in linear growth rates (cm/d). Two and three wave cycles differed (P < 0.05) with respect to the maximum diameter of both the first dominant follicle (1.51 +/- 0.24 vs 1.33 +/- 0.18 cm) and the ovulatory follicles (1.55 +/- 0.16 vs 1.34 +/- 0.13 cm). No relationship was found between dominant follicle development and the presence of either a CL or a previous dominant follicle in either ovary. Two and three wave cycles also differed with respect to the mean length of intervals between ovulation (22.27 +/- 0.89 vs 24.50 +/- 1.88 d; P < 0.01) and the mean length of luteal phases (10.40 +/- 2.11 vs 12.66 +/- 2.91 d; P < 0.05). These results demonstrate that buffalo have estrous cycles with 1, 2 or 3 follicular waves; that 2-wave cycles are the most common; and that the number of waves in a cycle is associated with the luteal phase and with estrous cycle length.     

A delay differential equation model of follicle waves in women

This article presents a mathematical model for hormonal regulation of the menstrual cycle which predicts the occurrence of follicle waves in normally cycling women. Several follicles of ovulatory size that develop sequentially during one menstrual cycle are referred to as follicle waves. The model consists of 13 nonlinear, delay differential equations with 51 parameters. Model simulations exhibit a unique stable periodic cycle and this menstrual cycle accurately approximates blood levels of ovarian and pituitary hormones found in the biological literature. Numerical experiments illustrate that the number of follicle waves corresponds to the number of rises in pituitary follicle stimulating hormone. Modifications of the model equations result in simulations which predict the possibility of two ovulations at different times during the same menstrual cycle and, hence, the occurrence of dizygotic twins via a phenomenon referred to as superfecundation. Sensitive parameters are identified and bifurcations in model behaviour with respect to parameter changes are discussed. Studying follicle waves may be helpful for improving female fertility and for understanding some aspects of female reproductive ageing.     

Patterns of expression of steroidogenic enzymes during the first wave of the ovine estrous cycle as compared to the preovulatory follicle

The expression patterns of steroidogenic enzymes in ovarian antral follicles at various stages of growth in a follicular wave have not been reported for sheep. Ovaries were collected from ewes (n=4-5 per group) when the largest follicle(s) of the first wave of the cycle, as determined by ultrasonography, reached (i) 3 mm, (ii) 4 mm, (iii) > or =5 mm in diameter or when there was a single (iv) preovulatory follicle in the last wave of the cycle, 12h after estrus detection. The expression pattern of steroidogenic enzymes was quantified using immunohistochemistry and grey-scale densitometry. The expression of CYP19 in the granulosa and 3beta-HSD and CYP17 in the theca increased (P<0.01) progressively from 3 to > or =5 mm follicles in the first wave of the cycle and was lower (P<0.01) in the preovulatory follicle compared to > or =5 mm follicles. However, the expression of 3beta-HSD in the granulosa increased (P<0.05) from 3 to > or =5 mm follicles and was maintained (P<0.05) at a high level in the preovulatory follicles. The amount of CYP19 in the granulosa of the growing follicles correlated positively (r=0.5; P<0.03) with the concurrent serum estradiol concentrations. We concluded that the expression pattern of steroidogenic enzymes in theca and granulosa of follicles growing in each wave in the ewe, paralleled with serum estradiol concentrations, with the exception that concentrations of 3beta-HSD in granulosa increased continuously from follicles 3mm in diameter to the preovulatory follicle.     

Ovarian antral follicular dynamics and their relationships with endocrine variables throughout the oestrous cycle in breeds of sheep differing in prolificacy.

Transrectal ultrasonography of ovaries was performed each day in non-prolific Western white-faced (n = 12) and prolific Finn ewes (n = 7), during one oestrous cycle in the middle portion of the breeding season (October-December), to record the number and size of all follicles > or = 3 mm in diameter. Blood samples collected once a day were analysed by radioimmunoassay for concentrations of LH, FSH and oestradiol. A cycle-detection computer program was used to identify transient increases in concentrations of FSH and oestradiol in individual ewes. Follicular and hormonal data were then analysed for associations between different stages of the lifespan of the largest follicles of follicular waves, and detected fluctuations in serum concentrations of FSH and oestradiol. A follicular wave was defined as a follicle or a group of follicles that began to grow from 3 to > or = 5 mm in diameter within a 48 h period. An average of four follicular waves per ewe emerged during the interovulatory interval in both breeds of sheep studied. The last follicular wave of the oestrous cycle contained ovulatory follicles in all ewes, and the penultimate wave contained ovulatory follicles in 10% of white-faced ewes but in 57% of Finn ewes. Transient increases in serum concentrations of FSH were detected in all animals and concentrations reached peak values on days that approximated to follicle wave emergence. Follicular wave emergence was associated with the onset of transient increases in serum concentrations of oestradiol, and the end of the growth phase of the largest follicles (> or = 5 mm in diameter) was associated with peak serum concentrations of oestradiol. Serum FSH concentrations were higher in Finn than in Western white-faced ewes during the follicular phase of the cycle (P < 0.05). There were no significant differences in serum concentrations of LH between Western white-faced and Finn ewes (P > 0.05). Mean serum concentrations of oestradiol were higher in Finn compared with Western white-faced ewes (P < 0.01). It was concluded that follicular waves (follicles growing from 3 to > or = 5 mm in diameter) occurred in both prolific and non-prolific genotypes of ewes and were closely associated with increased secretion of FSH and oestradiol. The increased ovulation rate in prolific Finn ewes appeared to be due primarily to an extended period of ovulatory follicle recruitment.     

Ovarian follicular populations and preovulatory enlargement in Booroola and control Merino ewes

To investigate the factors contributing to the different ovulation rates observed in two strains of sheep (Booroola 5.2, Merino 1.2), in-vivo monitoring of follicular kinetics followed by histological examination of both ovaries was performed during the late luteal and follicular phases. Ewes of both strains were either ovariectomized at Day 13, or had the 3 largest follicles of each ovary ink-labelled at Day 13 and were ovariectomized at Day 15, or had the 3 largest follicles of each ovary ink-labelled at Days 13 and 15 and were ovariectomized 16 h after the beginning of oestrus (N = 6 per time per strain). In another experiment, the age effects on the follicular populations of these two strains were also studied. There were 2-4 times more primordial follicles and 1.5-2 times more preantral follicles in the ovaries of Booroola than in control Merino ewes, although the number of antral follicles was the same. The percentage of normal follicles in this population was higher in Merino than Booroola ovaries. In Booroola ewes, there was no correlation between the number of antral follicles per ovary and the ovulation rate at the previous cycle (r = 0.22). This suggests that follicle numbers do not play a key role in the high ovulation rate of the Booroola strain. The number of follicles initiating growth from the primordial pool, the number of growing follicles disappearing at the preantral stage, the pattern of antrum development, granulosa cell multiplication and appearance of atresia differed between strains. The reasons for the high ovulation rate of the Booroola strain became clear when preovulatory enlargement was followed by ink labelling. An extended period of time during which recruitment of ovulatory follicles takes place, together with a low incidence of selection and the ability of the follicles to wait for ovulation are the features involved in this high ovulation rate.     

Relationships between FSH surges and follicular waves during the estrous cycle in mares

Individual follicles >/=15 mm were monitored daily by ultrasonography in 12 mares during the estrous cycle. Follicular waves were designated as major waves (primary and secondary) and minor waves based on maximum diameter of the largest follicle of a wave (major waves, 34 to 47 mm; minor waves, 18 to 25 mm). Dominance of the largest follicle of major waves was indicated by a wide difference (mean, 18 mm) in maximum diameter relative to the second largest follicle. Dominant follicles of primary waves (n=12) emerged (attained 15 mm) at a mean of Day 12 and resulted in the ovulations associated with estrus (ovulation=Day 0). The dominant follicle of a secondary wave (n=1) emerged on Day 2 and subsequently ovulated in synchrony with the dominant follicle of the primary wave, which emerged on Day 9. The largest follicles of minor waves (n=4) emerged at a mean of Day 5, reached a mean maximum diameter 3 days later, and subsequently regressed. There was a significant increase in mean daily FSH concentrations either 6 days (primary wave) or 4 days (minor waves) before the emergence of a wave. Mean concentrations of FSH decreased significantly 2 days after emergence of the primary wave. Divergence between diameter of the dominant and largest subordinate follicle of the primary wave was indicated by a significantly greater mean diameter of the dominant follicle than of the largest subordinate follicle 3 days after wave emergence and by the cessation of growth of the largest subordinate follicle beginning 4 days after the emergence of a wave. Surges of FSH were identified in individual mares by a cycle-detection program; surges occurred every 3 to 7 days. Elevated mean FSH concentrations over the 6 days prior to emergence of the primary wave was attributable to a significantly greater frequency of individual FSH surges before wave emergence than after emergence and to an increase in magnitude of peak concentrations of FSH associated with individual surges.     

Selection of a dominant follicle and suppression of follicular growth in heifers

The following aspects of follicle-stimulating hormone (FSH)-follicular relationships were studied in heifers: (1) the role of the decline in circulating levels of FSH in selection of a dominant follicle of a follicular wave; (2) the relationship of an FSH nadir (low levels between surges) to the absence of development of new follicles of a detectable diameter during the interim between the emergence of successive waves. A recombinant DNA-derived bovine FSH was used. Administration of bovine follicle-stimulating hormone (bFSH) for two days before the time of selection of the dominant follicle of the first post-ovulatory follicular wave delayed the time of divergence of the follicles into dominant and subordinates (first significant divergence: bFSH treatment before selection, Day 4.0; bFSH treatment after selection, Day 2.5; controls, Day 2.5: ovulation, Day 0). Significantly greater growth of the first and second largest subordinates occurred in the pre-selection group. A superovulatory dose of bFSH for 4 days with PGF₂-induction of luteolysis resulted in multiple ovulations when begun on Day 1 (before the expected time of follicle divergence; mean 2.8 ovulations per heifer) than when begun on Day 5 (after divergence; mean 1.0 ovulation per heifer). Administration of bFSH during the expected time (Days 5 and 6) of an FSH nadir did not alter the day of detectable emergence of the next follicular wave. Results supported the following hypotheses: (1) a decline in the wave-stimulating FSH surge is an integral component of the selection mechanism that results in the divergence into dominant and subordinate follicles; (2) the nadir between FSH surges does not account directly for the absence of the development of new follicles between the emergence of waves.     

Follicle and systemic hormone interrelationships during spontaneous and ablation-induced ovulatory waves in mares

The characteristics of ovulatory follicular waves were studied for spontaneous waves and waves induced during the next estrous cycle by ovarian follicle ablations and administration of PGF2alpha 10 days after ovulation in 21 mares. In the induced group, both the days of the FSH surge and day of deviation were more synchronized, LH concentrations were greater before and after deviation, estradiol concentrations were greater after deviation, and the ovulatory follicle grew at a faster rate (3.4+/-0.2 compared with 2.7+/-0.1 mm/day). The frequency of two dominant follicles/wave was not different between induced waves (7 of 21) and spontaneous waves (9 of 21), but both dominant follicles ovulated more frequently in induced waves (6 of 7 waves compared with 0 of 9).     

Ovarian function in ewes at the onset of the breeding season

Transrectal ultrasonography of ovaries was performed each day, during the expected transition from anoestrus to the breeding season (mid-August to early October), in six Western white-faced cross-bred ewes, to record ovarian antral follicles > or = 3 mm in size and luteal structures. Jugular blood samples were collected daily for radioimmunoassay (RIA) of follicle-stimulating hormone (FSH), oestradiol and progesterone. The first ovulation of the breeding season was followed by the full-length oestrous cycle in all ewes studied. Prior to the ovulation, all ewes exhibited a distinct increase in circulating concentrations of progesterone, yet no corpora lutea (CL) were detected and luteinized unovulated follicles were detected in only three ewes. Secretion of FSH was not affected by the cessation of anoestrus and peaks of episodic FSH fluctuations were associated with the emergence of ovarian follicular waves (follicles growing from 3 to > or = 5 mm). During the 17 days prior to the first ovulation of the breeding season, there were no apparent changes in the pattern of emergence of follicular waves. Mean daily numbers of small antral follicles (not growing beyond 3 mm in diameter) declined (P < 0.05) after the first ovulation. The ovulation rate, maximal total and mean luteal volumes and maximal serum progesterone concentrations, but not mean diameters of ovulatory follicles, were ostensibly lower during the first oestrous cycle of the breeding season compared with the mid-breeding season of Western white-faced ewes. Oestradiol secretion by ovarian follicles appeared to be fully restored, compared with anoestrous ewes, but it was not synchronized with the growth of the largest antral follicles of waves until after the beginning of the first oestrous cycle. An increase in progesterone secretion preceding the first ovulation of the breeding season does not result, as previously suggested, from the ovulation of immature ovarian follicles and short-lived CL, but progesterone may be produced by luteinized unovulated follicles and/or interstitial tissue of unknown origin. This increase in serum concentrations of progesterone does not alter the pattern of follicular wave development, hence it seems to be important mainly for inducing oestrous behaviour, synchronizing it with the preovulatory surge of luteinizing hormone (LH), and preventing premature luteolysis during the ensuing luteal phase. Progesterone may also enhance ovarian follicular responsiveness to circulating gonadotropins through a local mechanism.     

Characteristics of ovarian follicular dynamics throughout the estrous cycle of Egyptian buffaloes

Data of 56 normal and 9 abnormal estrous cycles were collected from 9 Egyptian buffaloes (Bublus bublis) to describe the follicular growth wave pattern. Heat was checked twice daily while, ovaries were scanned daily to monitor the patterns of follicular waves. Day of ovulation was determined when the largest follicle was replaced by corpus haemorrhgicum (CH). Number of waves/cycle, day of emergence of the follicular wave, characteristics of the dominant follicle and corpus luteum (CL) growth features were monitored. Buffaloes displayed mainly two types of follicular waves; two (46.4%) and three (53.6%). In cycles of three wave pattern, time of emergence of the 1st wave post-heat was longer (P < 0.05) and number of recruited follicles/wave were larger (P < 0.05) compared to the corresponding values of the two wave pattern. Number of recruited follicles in early follicular waves (1st or the 2nd) had larger number (P < 0.05) compared to the subsequent ones. Follicles that reached ovulation in both types of estrous cycle had shorter life-span (P < 0.05) than the previous ones. Life-span of CH, growing and regressed CL were 3.6 ± 0.6, 11.2 ± 0.8 and 4.4 ± 0.5 days, respectively with no difference in both types of follicular wave. Three types of ovarian disorders were observed. Follicular waves and CL growth features showed unique pattern for each individual. These results demonstrate that buffaloes display two main types of follicular waves with dominance of three wave type.