Waves Of Follicular Development Research Articles

The use of hormonal treatments to improve the reproductive performance of lactating dairy cows in feedlot or pasture-based management systems

Hormonal interventions have been used to increase the probability of estrous detection and insemination, and to increase pregnancy rates of dairy cattle under a variety of management systems. The present review addresses the basic principles of hormonal intervention and presents typical examples that illustrate the methodology. The hormones used to control the estrous cycle mimic the reproductive hormones found within the normal cow. Most estrous synchronization systems employ a method for controlling follicular wave development, promoting ovulation in anestrous cows, regressing the corpus luteum in cyclic cows, and synchronizing estrus and (or) ovulation at the end of treatment. A wide range of reproductive systems are in place on dairy farms. In most herds, a non-intervention period is practiced where postpartum cows are observed estrus estrus. Cows not observed in estrus are then treated. A number of studies in pasture-based and confinement systems have demonstrated net benefits of whole-herd synchronization. Despite the advantages of whole-herd reproductive programs, their uptake has been inconsistent globally. The benefits of a timed artificial insemination (AI) system increase under conditions of poor estrous detection rate and poor conception rate. The unpopular nature of timed AI programs in pasture-fed cows relates to high rates of estrous detection and conception for pasture-based dairying. Regardless of production system, some cows must be re-inseminated because they are not pregnant after first insemination. The presence of “phantom cows” (non-pregnant cows that do not return to estrus) creates a serious reproductive challenge for both pasture-based and confinement-style operations. Early pregnancy diagnosis and second insemination timed AI may reduce the effects of phantom cows on dairy herds. Fundamental research into anestrous, the hormonal control of the estrous cycle, and early pregnancy detection should elucidate new methods that can be used to strengthen reproductive programs on dairy farms.

The influence of the corpus luteum on ovarian follicular dynamics during estrous synchronization in goats

Ovarian follicular dynamics and fertility are unaffected by the presence or absence of a corpus luteum during synchronization of estrus with progestins in goats. On day 5 of the estrous cycle (estrus=day 0), a gestagen-containing sponge was inserted in the vagina for 11 days. To remove corpora lutea, one group of goats (CL−, n=41) received 7.5 mg of luprostiol on days 7 and 8 of the estrous cycle. The second group of goats retained the CL (CL+, n=38). Growth and development of follicles ≥4 mm in diameter were measured daily from onset of estrus to 2 days after subsequent ovulation in seven goats from each group, using rectal ultrasonography. Estrus was detected by the use of a reproductively sterilized buck and estrous does were subsequently mated. The number of waves of follicular development (CL −=3.57±0.2 versus CL+=3.14±0.14; P>0.05) did not differ between groups. The second wave of follicular development was present at the time of progesterone decline in the CL− group and neither its duration (CL−=4.8±0.4 versus CL+=5.6±0.7 days; P>0.05) nor the day of commencement of the third wave of follicular development (CL−=11.6±0.7 versus CL+=11.8±0.6; P>0.05) were altered by the concentration of endogenous progesterone. The pregnancy rate was similar between the two groups. (CL−=68.29% versus CL+=65.79%; P>0.05). Thus, in goats, ovarian follicular dynamics and fertility were not altered by the presence or absence of a corpus luteum during estrous synchronization.

Effects of medroxyprogesterone acetate (MAP) on ovarian antral follicle development, gonadotrophin secretion and response to ovulation induction with gonadotrophin-releasing hormone (GnRH) in seasonally anoestrous ewes

When ovulation is induced with gonadotrophin-releasing hormone (GnRH) in anoestrous ewes, a proportion of animals fail to form normal (full-lifespan) corpora lutea (CL). Progesterone treatment before GnRH prevents luteal inadequacy. It remains uncertain whether a similar effect, achieved with medroxyprogesterone acetate (MAP) from intravaginal sponges, is mediated by influences on growing ovarian follicles and/or secretion of gonadotrophic hormones, before and after GnRH treatment. Two experiments were performed, on 13 and 11 anoestrous Western white-faced ewes, respectively. Seven and six ewes, respectively, received MAP-containing sponges (60 mg) for 14 days; the remaining ewes served as untreated controls. To test the effect of timing of GnRH administration after pre-treatment with MAP-releasing sponges, GnRH injections (250 ng every 2 h for 24 h followed by a bolus injection of 125 μg of GnRH i.v.) were given either immediately (Experiment 1) or 24 h after sponge removal in the treated ewes (Experiment 2). Ovarian follicular dynamics (follicles reaching ≥5 mm in size) and development of luteal structures were monitored using transrectal ultrasonography. In Experiment 1, the mean ovulation rate (0.7±0.3 and 1.0±0.4) and proportion of ovulating ewes (57 and 67%, respectively) did not vary ( P>0.05) between MAP-treated and control ewes. Normal (full-lifespan) CL were detected in 29% of treated and 67% of control ewes ( P>0.05). In Experiment 2, the mean ovulation rate (2.3±0.2 and 1.2±0.6; P<0.05) and percentage of ewes with normal (full-lifespan) CL (100 and 40%, respectively; P<0.10) were greater in the treated compared to control ewes. In Experiment 1, the mean peak concentration of the GnRH-induced LH surge was lower ( P<0.05) in MAP-treated than in control ewes. There were no significant differences between MAP-treated and control ewes in the characteristics of follicular waves, mean daily serum FSH concentrations, and secretory parameters of LH/FSH, based on intensive blood sampling conducted 1 day before sponging and 1 day before sponge removal. It is concluded that treatment with MAP has no effect on the tonic secretion of LH/FSH or follicular wave development in anoestrous ewes. However, the GnRH-stimulated LH discharge was attenuated in the ewes that received MAP-impregnated sponges for 14 days and were treated with GnRH immediately after sponge withdrawal. Ovulatory response and CL formation were increased when GnRH was administered 24 h after sponge removal.

Ovulatory Cycle Incorporates Two to Three Waves of Follicular Development

Ovulatory Cycle Incorporates Two to Three Waves of Follicular Development

Ovulatory Cycle Incorporates Two to Three Waves of Follicular Development

Ovulatory Cycle Incorporates Two to Three Waves of Follicular Development

Proteolysis of insulin-like growth factor binding proteins -4 and -5 in bovine follicular fluid: implications for ovarian follicular selection and dominance.

Dominant follicles are characterized by low levels of low molecular weight IGF binding proteins (IGFBPs) and by proteolytic activity against IGFBP-4 and -5. To examine the hypothesis that proteolysis of IGFBP-4 and -5 plays a critical role in selection of the dominant follicle, we isolated follicles at various stages during the first wave of follicular development during the bovine estrous cycle, using ultrasonography to follow follicular growth. Ovariectomies were performed before divergence in follicular size (group 1; largest follicle, approximately 7 mm in diameter), at about the expected time of size divergence (group 2; largest follicle, approximately 8 mm) or after a dominant follicle was clearly present (group 3; largest follicle, > or =9 mm). The four largest follicles (F1-F4) were dissected and concentrations of steroids, IGFBPs and free IGF-I and levels of proteolytic activity for IGFBP-4 and -5 in the follicular fluid were determined. Follicles in group 1 did not differ significantly in size or estradiol concentrations, but levels of proteolytic activity against IGFBP-4 and -5 were higher in F1-F2 than in F3-F4. However, in group 2 the largest follicle (F1) had higher estradiol, free IGF-I, and IGFBP-4 and -5 proteolytic activity than F2-F4, whereas only slight (dissected) or no (ultrasound) differences in diameters could be detected. Differences between F1 and F2-F4 in diameter, estradiol, free IGF-I, and IGFBP-4 and -5 proteolytic activity were even greater in group 3. In addition, the hormonal regulation of IGFBP-4 and -5 proteolysis was evaluated in vivo by injecting heifers with small doses of recombinant bovine FSH to induce codominant follicles (group 4). The induced codominant follicles were larger and had higher IGFBP-4 and -5 proteolytic activity than subordinate follicles. The results suggest that follicular selection is a progression of changes starting with acquisition of an FSH-inducible IGFBP-4/-5 proteolytic activity, leading to an increase in intrafollicular concentration of free IGF-I that, in turn, synergizes with FSH to promote greater estradiol production by the follicle destined for dominance.

Pattern and manipulation of follicular development in Bos indicus cattle

Bos indicus cattle are widespread in tropical regions due to their adaptation to these environments. Although data on reproductive performance have indicated both inferior and superior results for B. indicus cattle, there is little doubt that B. indicus cattle are superior than Bos taurus cattle when they are both kept in tropical or subtropical environments, where stressors like hot temperatures, humidity, ectoparasites and low quality forages are greater. Reproductive endocrinology and oestrus behaviour of the B. indicus cattle have been studied for over 30 years; however, the application of technologies such as real time ultrasonography and Heat-Watch systems has expanded our knowledge on the ovarian follicular-wave dynamics during the oestrous cycle and the time of ovulation. Ovarian follicular dynamics in B. indicus cattle is characterised by the occurrence of two, three or sometimes four waves of follicular development. While dominance is similar to that in B. taurus cattle, maximum diameters of the dominant follicle and CL are smaller than those reported in B. taurus and are probably due to a lower capacity for LH secretion than in B. taurus. Duration of oestrus is approximately 10 h and the interval from oestrus to ovulation is about 27 h. However, the variability in response to prostaglandin F2α (PGF) treatments and the difficulty for oestrus detection in B. indicus cattle have limited the widespread application of artificial insemination (AI) and emphasizes the need for treatments that control follicular development and ovulation. Follicular-wave development in B. indicus cattle can be controlled mechanically by ultrasound-guided follicle ablation, or hormonally by treatments with GnRH or oestradiol and progestogen/progesterone in combination. Treatments with GnRH plus PGF and a second GnRH (synchronization protocol known as Ovsynch) or oestradiol benzoate (known as GPE) have resulted in acceptable pregnancy rates after fixed-time AI (FTAI) in cycling cows, but results were lower in heifers and cows in postpartum anoestrus. Alternatively, treatments with oestradiol and progestogen/progesterone releasing devices resulted in synchronous emergence of a new follicular wave, and a second oestradiol or GnRH treatment after device removal resulted in synchronous ovulation and acceptable pregnancy rates to FTAI. Furthermore, oestradiol and progesterone treatments combined with eCG (given at the time of device removal) increased pregnancy rates in suckled B. indicus cows and may be useful for the treatment of cows in postpartum anoestrus. In summary, exogenous control of luteal and follicular development facilitates the application of assisted reproductive technologies in B. indicus cattle by offering the possibility of planning AI programs without the necessity of oestrus detection and without sacrificing the overall results.

Estradiol benzoate delays new follicular wave emergence in a dose-dependent manner after ablation of the dominant ovarian follicle in cattle.

Administration of estradiol benzoate (EB) induces atresia of the dominant follicle (DF) in the ovaries of cattle within 36 h but emergence of a new wave of follicular development is delayed by 3-5 days. The present study investigated the role of EB in determining timing of emergence of a new follicular wave after removing the influence of the DF. At 6.4+/-0.2 days after ovulation in Angus and Angus/Simmental cattle (n=26), aged 4.9+/-0.6 years and weighing 634+/-20 kg, all ovarian follicles > or =5mm in diameter were aspirated with a 17-gauge needle using an ultrasound-guided transvaginal approach (Day 0 or Hour 0) and animals immediately received 0 (0EB), 1 (1EB), 2 (2EB) or 4 (4EB) mg EB i.m./500 kg body weight (n=6 or 7 per treatment). Ovarian structures were monitored by ultrasonography on a daily basis until emergence of a new wave of follicular development. Concentrations of estradiol (E2) were different among all treatments between Hours 24 and 72, increasing (P<0.01) with greater doses of EB administered. Hour of peak follicle-stimulating hormone (FSH) was 29.3+/-4.0, 53.3+/-4.5, 81.1+/-15.5, and 91.4+/-8.2 for the 0EB, 1EB, 2EB, and 4EB treatments, respectively, and emergence of a new wave of follicular development occurred on Days 1.5+/-0.2, 3.3+/-0.3, 4.0+/-0.6 and 4.4+/-0.4, respectively. Timing of peak FSH and emergence of a new wave of follicular development was earliest (P<0.05) in the 0EB treatment, similar (P>0.1) among the 1EB and 2EB treatments, and most delayed (P<0.05) in the 4EB treatment when compared to the 0EB or 1EB treatments. The overall mean interval from peak FSH to emergence of a new wave of follicular development was 15.7+/-3.3 h and was not affected by treatment. Concentrations of E2 at 24 h before new emergence were not different among EB-treated animals (20.2+/-5.5 pg/ml), but lower (P<0.01) in the 0EB treatment (1.6+/-0.2 pg/ml). In a dose-dependent manner, EB delayed the pre-emergence surge in FSH that stimulates new follicular development after the DF has ceased to be functional. The importance of using an 'optimal' dose of EB in hormonal regimens using this agent to strategically regulate follicular development is emphasized by the outcomes of this study.

Ovarian Dynamics and Their Associations with Peripheral Concentrations of Gonadotropins, Ovarian Steroids, and Inhibin During the Estrous Cycle in Goats1

Ovarian changes determined by daily transrectal ultrasound and its relationship with FSH, LH, estradiol-17beta, progesterone, and inhibin were investigated in six goats for three consecutive interovulatory intervals. Estrous cycles were synchronized using two injections of prostaglandin F2alpha analogue 11 days apart. All follicles 3 mm or greater in diameter and corpora lutea were measured daily. A follicular wave was defined as one or more follicles growing to 5 mm or greater in diameter. The day that the follicles reached 3 mm in diameter was defined as the day of wave emergence, and the first wave after ovulation was defined as wave 1. During the interovulatory interval (mean +/- SEM, 21.3 +/- 0.4 days; n = 18), follicular waves emerged at 0.3 +/- 0.5, 6.5 +/- 0.2, and 12.1 +/- 0.4 days for wave 1, wave 2, and wave 3, respectively, in goats with three waves of follicular development and at -0.6 +/- 0.3, 4.7 +/- 0.2, 9.4 +/- 0.5, and 13.4 +/- 0.5 days for wave 1, wave 2, wave 3, and wave 4, respectively, in goats with four waves of follicular development (Day 0 = the day of ovulation). The mean diameter of the largest follicle of the ovulatory wave was significantly larger than those of the largest follicles of the other waves. Corpora lutea could be identified ultrasonically at Day 3 postovulation and attained 12.1 +/- 0.3 mm in diameter on Day 8. Transient increases in plasma concentrations of FSH were detected around the day of follicular wave emergence. The level of FSH was negatively correlated with that of inhibin. These results demonstrated that follicular waves occurred in goats and that the predominant follicular wave pattern was four waves with ovulation from wave 4. These results also suggested that the emergence of follicular waves was closely associated with increased secretion of FSH.

Influence of GnRH analogue (fertirelin acetate) doses on synchronization of ovulation and fixed-time artificial insemination in lactating dairy cows

The effect of using a dose of 50 μg rather than 100 μg fertirelin in an ovulation/fixed-time insemination protocol for Holstein–Friesian dairy cows was investigated in three experiments. In each experiment, fertirelin was administered at the beginning of the protocol followed 7 days later by 500 μg cloprosterol. Two days later, a second dose of fertirelin was given and AI performed 16–19 h later regardless of the incidence of behavioral oestrus. The effect on conception rate was studied in experiment 1 using 114 postpartum anoestrus cows. There was no significant difference in the age, parity or number of days after parturition in each treatment groups. The conception rate did not differ between the 50 μg fertirelin group (61.1%; n=72) and the 100 μg group (59.5%; n=42; NS). In experiment 2, a further 12 cows at 40–60 days postpartum were treated with 100 or 50 μg fertirelin ( n=6 per dose) with treatment commencing in the follicular or luteal phase of the oestrous cycle. The plasma concentration of luteinizing hormone (LH) reached similar peaks of over 5 ng/ml 120 min after the intramuscular administration of fertirelin in both groups. There were no significant differences in LH levels between treatments or phase of the oestrous cycle when treatment commenced. Doses of 50 and 100 μg fertirelin were compared in experiment 3 using 17 cows to study follicular wave development and synchronization by transrectal ultrasonography, conception rate and corpus luteum function. There were no significant differences between treatments for these factors. It was concluded that using a dose of 50 μg fertirelin enabled the drug costs to be reduced without affecting the efficiency of a synchronization of ovulation/fixed-time AI protocol for dairy cows.

Ovarian follicular dynamics and endocrine profiles in Polwarth ewes with high and low body condition

AbstractIn this investigation we tested the hypothesis that static body condition (BC) of the ewe affects oestradiol and FSH with implications for subsequent follicular growth and turn-over. Sixteen Polwarth ewes were selected from a flock according to their BC score (scale: 1 emaciated; 5 obese). High BC (HBC) ewes (no. = 8) had a BC score of 4·1 (s.e. 0·1) and low BC (LBC) ewes (no. = 8) had a BC score of 1·9 (s.e. 0·1). Daily ultrasound examinations were performed and blood samples for progesterone, oestradiol and follicle-stimulating hormone (FSH) determination were collected. All HBC ewes (8/8) exhibited three waves of follicular development, while four LBC ewes (4/8) had two waves and the other four (4/8) had three waves of follicular development (P ≤ 0·05) during the interovulatory period. Overall, the emergences of 33 out of 44 follicular waves were preceded by significant increases in FSH concentrations. Maximum FSH concentrations were detected 0·9 ± 0·2 days before wave emergence. Oestradiol concentrations increased significantly during the growing phase in 38 out of 44 large follicles. A negative correlation between oestradiol and FSH was observed in HBC ewes. A similar inhibitory effect of oestradiol on FSH was observed in LBC ewes, irrespective of whether they developed two or three follicular waves. However, a longer period with high FSH was needed to promote the emergence of the second follicular wave in two-wave LBC ewes. Four HBC ewes had twin ovulations but no LBC ewes did (P ≤ 0·05). In HBC ewes, the follicular phase was characterized by lower oestradiol (6·5 (s.e. 1·0) pmol/l) but higher mean FSH concentrations (2·4 (s.e.0·4) μg/l) than in LBC ewes (8·9 (s.e. 1·2) pmol/l and 2·0 (s.e. 0·3) μg/l, respectively; P ≤ 0·05). The present results suggest that BC influences the pattern of follicular dynamics through changes in the endocrine milieu. Higher FSH concentrations during the follicular phase in HBC ewes, which allowed an extended period of follicular recruitment from a significantly larger pool of small antral follicles could explain the higher ovulation rate observed in this group.

The control of follicular wave development for self-appointed embryo transfer programs in cattle

Our expanding knowledge of the control of follicular wave dynamics during the bovine estrous cycle has resulted in renewed enthusiasm for the prospects of precisely controlling the follicular and luteal dynamics and finely controlling the time of ovulation. Follicular wave development can be controlled mechanically by ultrasound-guided follicle ablation or hormonally by treatments with GnRH or estradiol and progestogen/progesterone in combination. Treatment of cattle with GnRH in combination with prostaglandin F 2α (PGF) 7 d later and a second GnRH 48 h after PGF (known as Ovsynch) has resulted in acceptable pregnancy rates after fixed-time AI in lactating dairy cows and in recipients in which embryos were transferred without estrus detection. Alternatively, treatments with estradiol and progestogen/progesterone-releasing devices resulted in synchronous emergence of a new follicular wave and, when a second estradiol treatment was given 24 h after device removal, synchronous ovulation and high pregnancy rates to fixe-time AI. Self-appointed embryo transfer (without estrus detection) using estradiol and progesterone treatments have resulted in pregnancy rates comparable with those obtained with recipients transferred 7 d after estrus. Furthermore, estradiol and progesterone treatments combined with PGF and eCG (given 1 d after the expected time of wave emergence) have resulted in high rates of recipients selected for transfer (84.6%) and an overall pregnancy rate of 48.7% (recipients pregnant/recipients treated). Estradiol and progestogen/progesterone treatments have also been widely used for self-appointed superstimulation protocols with equivalent embryo production to that of donor cows superstimulated using the traditional approach beginning 8 to 12 d after estrus. In summary, exogenous control of luteal and follicular development facilitates the application of assisted reproductive technologies in cattle by offering the possibility of planning the superstimulation of donors and synchronization of recipients at a self-appointed time, without the necessity of estrus detection and without sacrificing results.

Effects of monensin and forage:concentrate ratio on feed intake, endocrine, and ovarian function in beef heifers

The objective of this study was to determine the effects of monensin (M) and altered forage:concentrate ratio (F:C) on feed intake, hormonal profiles, and ovarian function in beef heifers. Four ruminally cannulated heifers were maintained in metabolism crates and fed twice daily. Diets contained 0 or 200 mg per day M in diets of 70:30 (high forage, HF) or 30:70 (high concentrate, HC) F:C. Diets were limit fed to achieve isocaloric intake of calculated ME. Monensin supplementation had no effect on feed intake, but heifers fed HF had greater refusals compared with heifers fed HC ( P<0.005). Serum insulin, progesterone, and estradiol concentrations did not differ between diets or treatments ( P>0.10). Diet and/or treatment tended to alter the size or number of follicles on either or both ovaries. Monensin resulted in more waves of follicular development per estrous cycle with HF diets, but fewer with HC diets ( P<0.07). The interovulatory interval, lifespan of the corpus luteum, and the size of the dominant follicle in the first two follicular waves were not different due to diet or treatment. However, the HC diet tended to produce a larger ovulatory follicle compared with the HF diet (HF=12.25, HC=13.5; P<0.08). We conclude that feeding monensin affects estradiol secretion and a higher proportion of concentrates alters ovarian dynamics, which could affect fertility in beef heifers.

Ovarian follicular dynamics of five-eighths Girolando cows.

The objective of this study was to examine the follicular dynamics of five-eighths Girolando cows by observing the number of follicular development waves, days of emergence of those waves, diameters of the dominant and largest subordinate follicles and the processes of follicular selection and dominance. Ovarian follicle dynamics were monitored for 24 oestrous cycles in 12 cows, of 4 to 10 years of age, presenting regular oestrous cycles and with body scores between 3 and 4. Ovaries were observed daily for two consecutive oestrous cycles and follicles were measured with ultrasonographic equipment. The dominant follicle was considered to be that which presented a diameter > or = 10 mm for three consecutive days and on the day of wave emergence, when a group of follicles measuring 3-5 mm in diameter would appear. Of the 24 cycles monitored, 62.5% presented two waves of follicular development and 37.5% presented three waves. The cycles presenting two waves had an average duration of 20 days, with the emergence of the waves on days 1 and 9, whereas the cycles presenting three waves had an average duration of 22 days, with emergences on days 1, 10 and 16. For cycles with two waves, both the first and second dominant follicles reached an average size of 13.8 mm, with the first regressing on day 10 and the second ovulating around day 20. For cycles with three waves, the dominant follicles of the first and second waves reached maximum diameters of 11.8 and 12 mm, respectively, with the first regressing on day 11 and the second regressing on day 17. The third dominant follicle reached a maximum diameter of 12.4 mm on day 20, and ovulated on day 22. These results lead to the conclusion that the follicular dynamics of five-eighths Girolando cows are characterized by the presence of two to three waves of follicular growth.

Alterations in intrafollicular regulatory factors and apoptosis during selection of follicles in the first follicular wave of the bovine estrous cycle.

Changes in follicular fluid (FF) concentrations of estradiol, inhibin forms, and insulin-like growth factor binding proteins (IGFBPs), percentage of apoptotic granulosa cells (%A), and follicular size for individual follicles in a growing cohort were determined throughout the first wave of follicular development during the bovine estrous cycle and related to FSH decline. Four groups of heifers (n = 31) were ovariectomized between Days 1.5 and 4.5 of the estrous cycle at 5 +/- 1, 33 +/- 2, 53 +/- 1, and 84 +/- 2 h after the periovulatory peak in FSH concentrations. Follicles > or = 2.5 mm were dissected, measured, and FF aspirated. The five largest follicles were ranked based on their diameter (F1 to F5). Diameters of F1 to F5 were positively correlated with interval from FSH peak (r > or = 0.6, P < 0.05). Five hours after the FSH peak, follicular diameter and FF concentrations of estradiol, inhibins, and IGFBPs were similar for F1 to F5. From 5 to 33 h, amounts of the six precursor inhibin forms (> or = 48 kDa) increased (P < 0.05) in F1 follicles. The IGFBPs in F1 follicles remained low at all time periods. At 33 h, amounts of IGFBP-4 and -5 were higher (P < 0.05) in F4 and F5 compared with F1 follicles. At 84 h, IGFBP-2, -4, and -5 were increased (P < 0.05) in F3, F4, and F5 compared with F1. At 5, 33, or 53 h, %A was not different between follicles in any size class. At 84 h %A was increased (P < 0.05) in follicles <6 mm in diameter. However, at that time, %A did not differ between the selected DF and the largest subordinate follicle. For individual heifers, the selected DF at 84 h was largest in size, highest in estradiol, and lowest in IGFBP-2 and -4. The F1 follicle had highest estradiol in 23 of 27 heifers irrespective of stage of the wave and lowest IGFBP-4 in 19 of 21 heifers from 33 h. We concluded that the earliest intrafollicular changes that differentiate a dominant-like follicle from the growing cohort are enhanced capacity to produce estradiol and maintenance of low levels of IGFBPs.

Kinase pathways in dominant and subordinate ovarian follicles during the first wave of follicular development in sheep

The mechanism by which one or more dominant ovarian follicles continue development while other subordinate follicles regress is not known. The mitogen activated protein kinases (MAPKs) are a group of kinases that are activated by hormonal factors and form a cascade of processes that regulate cell growth, division and differentiation. The aim of the present experiment was to characterise the presence of the MAPKs, Erk 1/Erk 2 and Akt in healthy dominant follicles and regressing subordinate follicles. Following in vivo monitoring of ovarian follicle development, three ewes were ovariectomised and the follicular fluid and follicle wall (theca and granulosa cells) saved from the dominant and largest subordinate follicle. The dissected diameter and follicular fluid oestradiol concentration of the dominant follicle was larger ( P<0.01) than the largest subordinate follicle (6.5±0.0 mm and 41.3±4.9 ng/ml versus 4.7±0.3 mm and 0.6±0.4 ng/ml). Western blot analyses showed that there was more Akt (202.7±6.4 versus 59.6±32.7 units; P<0.05) and Erk 1/Erk 2 (104.5±10.6 versus 0.3±0.2 units; P<0.01) present in follicle wall samples from the dominant compared to the largest subordinate follicles. Phosphorylated forms of Akt and Erk 1/Erk 2 were detected in samples from dominant but not subordinate follicles. We suggest that signal transduction pathways involving Akt and Erk 1/Erk 2 may play an important role in determining the outcome of ovarian follicle growth and development in sheep.

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 ≥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.

Ovarian antral follicular dynamics and their associations with peripheral concentrations of gonadotropins and ovarian steroids in anoestrous Finnish Landrace ewes.

Daily transrectal ultrasonography of ovaries was done in seven Finn ewes during three 17-day periods from May to July. Blood samples were collected each day for estimation of the serum follicle-stimulating hormone (FSH), oestradiol and progesterone concentrations, and also every 15 min for 6 h, halfway through each period of ultrasonographic examination, to determine the patterns of gonadotropic hormone secretion. Four ewes ceased cycling from March to mid-April (ewes entering anoestrus early) and three in May (ewes entering anoestrus late). In all ewes cyclicity resumed during the period from mid-August to mid-September. The growth of ovarian antral follicles to periovulatory sizes of >/=5 mm in diameter was seen at all stages of anoestrus. An average of four waves of follicular development (follicles growing from 3 to >/=5 mm in diameter before regression) with a periodicity of 4 days were recorded during each of the three scanning periods. There was a close temporal relationship between days of follicular wave emergence and peaks of successive FSH fluctuations. Ewes entering anoestrus late exceeded ewes that became anoestrus early in numbers of large (>/=5 mm in diameter) ovarian antral follicles and maximum follicle diameter. Peak concentrations of transient FSH increases were higher (P<0.05) in ewes entering anoestrus late than in ewes entering anoestrus early. The secretion of luteinising hormone, (LH; mean and basal level, and LH pulse frequency, but not amplitude) was lowest during the month of June in all ewes. Oestradiol production was markedly suppressed throughout anoestrus. Peaks of progesterone secretion appeared to occur at regular intervals and were associated with the end of the growth phase of the largest follicles of sequential waves. In conclusion, the growth of ovarian follicles to ostensibly ovulatory diameters is maintained throughout anoestrus in Finn ewes and periodic emergence of follicular waves is correlated with an endogenous rhythm of FSH secretion. The present study also provides evidence for the inverse relationship between the time of the onset of seasonal anoestrus and the number and size of antral follicles developing throughout anoestrus in Finn ewes, and indicates that differences exist in both the secretion of and ovarian responsiveness to gonadotropic hormones among early and late anoestrous ewes.

Use of a small dose of estradiol benzoate during diestrus to synchronize development of the ovulatory follicle in cattle.

We tested the hypothesis that a small dose of estradiol benzoate (EB) at the midstage of the estrous cycle in cattle would synchronize the subsequent pattern of ovarian follicular development, estrus, and ovulation. Nonlactating Friesian cows received either 1 mg of EB i.m. on d 13 of the estrous cycle (T; n = 12; estrus = d0) or served as untreated controls (C; n = 12). Their ovaries were examined daily with transrectal ultrasonography from d 7, and blood samples were collected 0, 2, 4, 8, 24, and 48 h after treatment on d 13. Plasma concentrations of estradiol-17beta were elevated to 12 pg/mL during the initial 24 h following treatment, compared with a baseline of 1 pg/mL in untreated controls (P < .001). Progesterone concentrations in cows of the T group declined between 24 and 48 h after treatment (-3.2 +/- .5 ng/mL) compared with little change in concentrations of progesterone in cows of the C group at this time (P < .01). This difference was coincident with an earlier time to regression of the corpus luteum in cows of the T group. Disregarding treatment groups, the second dominant follicle of the estrous cycle (DF2) emerged on d 10.6 +/- .3 and was 9.4 +/- .4 mm in diameter on d 13. Further growth of the DF2 was halted by EB treatment on d 13. Cessation of growth occurred irrespective of whether the DF2 was in the early or late growth phase, and a new follicular wave emerged 4.5 +/- .2 d later. The dominant follicle from this wave (DF3) ovulated 5 d after emergence in most cases. During the estrous cycle of every cow in the T group, there were three waves of follicular development (3-wave), whereas the ratio of 2:3 waves of follicular development in cows of the C group was 1:3. Consequently, the interval from emergence to ovulation of the ovulatory dominant follicle in cows of the C group ranged from 3 to 11 d. The dynamics of ovarian follicular wave development during the estrous cycle can be strategically manipulated by treating with a small dose of EB to synchronize proestrous development of the ovulatory follicle.

Expression and Function of Fas Antigen Vary in Bovine Granulosa and Theca Cells During Ovarian Follicular Development and Atresia1

Fas antigen is a receptor that triggers apoptosis when bound by Fas ligand (FasL). A role for Fas antigen in follicular atresia was studied in follicles obtained during the first wave of follicular development during the bovine estrous cycle (estrus is Day 0). Granulosa and theca cells were isolated from healthy dominant follicles and the two largest atretic subordinate follicles on Day 5, atretic dominant follicles on Days 10-12, and preovulatory follicles on Day 1. Fas antigen mRNA levels were highest in granulosa cells from subordinate as compared to other follicles, and lowest in theca cells from healthy Day 5 dominant as compared to other follicles. FasL alone had no effect on viability of granulosa or theca cells but became cytotoxic in the presence of interferon-gamma (IFN). IFN has been shown to induce responsiveness to Fas antigen-mediated apoptosis in other cell types. In the presence of IFN, killing of granulosa cells by FasL was greater in subordinate compared to healthy dominant follicles on Day 5, did not differ between healthy and atretic dominant follicles, and was similar in theca among all follicles. Granulosa cells from preovulatory follicles, which had been exposed to the LH surge in vivo, were completely resistant to FasL-induced killing. In summary, Fas antigen expression, and responsiveness to Fas antigen-mediated apoptosis, vary during follicular development.