Ovarian follicular development in Holstein cows following synchronisation of oestrus with oestradiol benzoate and an intravaginal progesterone releasing insert for 5-9 days and duration of the oestrous cycle and concentrations of progesterone following ovulation

The aim of this study was to determine if the duration of treatment with an intravaginal progesterone releasing insert (IVP(4)) after treatment with oestradiol benzoate (ODB) at the time of insertion and 24 h after removal would affect selected variables including: size of ovarian follicles at the time of removal of inserts, diameter of ovulatory follicles, plasma concentrations of progesterone following ovulation, and duration of the following oestrous cycle. Characteristics of oestrus at a synchronised and spontaneous oestrus were also monitored. Non-lactating Holstein cows were synchronised with an IVP(4) for 5 (n = 10), 7 (n = 10), 8 (n = 9) or 9 (n = 9) days together with injections of ODB at device insertion (2 mg) and 24 h after removal (1 mg). Ultrasonography showed no significant effect of treatment on the day of emergence of preovulatory follicles relative to the day of removal of inserts (overall mean = -4.22 +/- 0.58; P = 0.15) for cows that ovulated within 120 h insert removal (n = 36). Treatment with ODB and an IVP(4) for 5 days reduced the diameter of preovulatory follicles at the time of removal of inserts and for the following 2 days compared to cows treated for 7-9 days (mean difference 2.56 +/- 1.15 mm; P = 0.033) but did not reduce the diameter of the ovulatory follicle (P = 0.21). Day of emergence relative to removal of inserts was associated with the diameter of the ovulatory follicle (R2 = 0.69; P < 0.001). Concentrations of progesterone and the diameter of the corpus luteum following ovulation were not affected by treatment (P > 0.20), but were affected by the diameter of the ovulatory follicle (P < 0.01). Diameter of the ovulatory follicle did not affect interoestrous and interovulatory intervals (P > 0.40). We conclude that treatment with an IVP(4) for 5 compared to 7-9 days with ODB administered at device insertion, and 24 h after removal reduced the diameter of preovulatory follicles at the time of removal of the insert but did not reduce the diameter of the ovulatory follicle or concentrations of progesterone in plasma. Emergence of preovulatory follicles closer to the time of removal of inserts reduced the diameter of the ovulatory follicle when oestrus was induced with ODB. Ovulation of smaller follicles reduced concentrations of progesterone in plasma following ovulation but did not affect oestrous cycle duration.     

Effects of weaning on concentrations of inhibin in follicular fluid and plasma of sows.

Changes in plasma and follicular fluid concentrations of inhibin were examined in sows after weaning at 28-32 days post partum. From 0 to 48 h after weaning, inhibin concentrations were 200-300 times higher in follicular fluid from small (less than 4 mm) and medium-large (greater than or equal to 4 mm) follicles than in ovarian venous plasma. Inhibin concentrations increased in follicular fluid from medium-large follicles at 24 and 48 h after weaning; concentrations in ovarian venous plasma were positively correlated with the number of medium-large follicles (r = 0.40) and with ovarian venous plasma concentrations of oestradiol (r = 0.61). Blood samples were collected for 30 days from sows (n = 6) that exhibited oestrus within 5 days after weaning and from sows (n = 5) that remained anoestrous for 11 days after weaning. Plasma inhibin concentrations rose in oestrous and anoestrous sows by 12 h and continued to rise for 60 h after weaning. Plasma inhibin concentrations rose further and were higher at 3.5-4.5 days after weaning in oestrous sows than in sows that remained anoestrous. After oestrus, plasma inhibin concentrations declined. At weaning, plasma concentrations of follicle-stimulating hormone (FSH) were higher in sows that subsequently exhibited oestrus than in sows that remained anoestrous. After weaning, plasma concentrations of FSH declined in both groups, reached a nadir at 2.5 days, and increased gradually in anoestrous sows; oestrous sows exhibited an FSH surge at oestrus. Plasma FSH returned to preweaning concentrations in both groups of sows at Days 7-8.(ABSTRACT TRUNCATED AT 250 WORDS)     

Inhibin production in vitro by granulosa cells from Booroola ewes which were either homozygous or non-carriers of a fecundity gene influencing their ovulation rate

The production of inhibin by granulosa cells was studied in vitro using cells from follicles of various sizes and health. Follicles were recovered on Days 10-13 of the oestrous cycle, from Booroola x Romney ewes which were homozygous (FF) carriers or non-carriers (++) of the fecundity (F) gene. Inhibin was measured using a bioassay based on the suppression of follicle-stimulating hormone (FSH) output by cultured pituitary cells from ovariectomized Romney ewes and, in some instances, for comparative purposes, by radioimmunoassay also. Geometric mean inhibin production by granulosa cells from nonatretic follicles increased with increasing follicle diameter, during the first 24 h of culture, for both genotypes. The geometric mean production of inhibin by cells from nonatretic 3-4.5 mm diameter FF follicles (the largest follicles found in FF ewes), was significantly higher (P less than 0.05) than that by cells from non-atretic 3-4.5 mm diameter ++ follicles, but similar to that of cells from non-atretic greater than or equal to 5 mm diameter ++ follicles. The production of oestradiol-17 beta by cells cultured in the presence of testosterone (1 microgram/ml) followed a pattern similar to cellular inhibin production. There was a positive linear correlation between inhibin and oestradiol-17 beta production during the first 24 h of culture, for both genotypes. In addition to acting as a substrate for oestradiol-17 beta synthesis, testosterone generally had a slight, stimulatory effect on inhibin production.(ABSTRACT TRUNCATED AT 250 WORDS)     

Concentrations of inhibin,progesterone and oestradiol in fluid from dominant and subordinate follicles from mares during spring transition and the breeding season

Dominant and subordinate follicles were collected from mares on the day after the dominant follicle reached 30 mm in diameter, to investigate regulation of folliculogenesis during spring transition and the breeding season. Concentrations of oestradiol-17beta, progesterone and inhibin A, but not inhibin isoforms with pro- and alpha C-immunoreactivity, were significantly higher in preovulatory follicles than in dominant anovulatory transitional follicles. Steroidogenic activity was regained gradually in the dominant follicles of successive anovulatory waves through spring transition. The dominant follicles, during both spring transition and cyclicity, contained higher concentrations of oestradiol, progesterone and inhibin A, but not inhibin pro- and alpha C-isoforms, than subordinate follicles. The results indicate that high follicular levels of oestradiol, progesterone and inhibin A are associated with continued follicle growth and ovulation. The low concentrations of oestradiol and progesterone in transitional follicles indicate that the deficiency in steroidogenesis exists early in the steroidogenic pathway. The similarity in patterns of follicular hormones in spring transition and during cyclicity strongly suggests that the mechanism of dominance is the same in both types of follicle.     

Hormonal changes during luteal regression in farmed fallow deer, Dama dama

Concentrations of progesterone, oxytocin and PGFM (pulmonary metabolite of PGF-2 alpha) were measured in plasma from peripheral blood samples collected from 5 fallow does every hour or 2 h for 12-h periods on Days 15-20 inclusive of the oestrous cycle (i.e. luteolysis). For 3 does that exhibited oestrus on Day 21, plasma progesterone concentrations fluctuated between 3 and 10 ng/ml on Days 15-18 inclusive. Thereafter, values declined progressively to attain minimum concentrations of less than 0.05 ng/ml on Day 20. Basal concentrations of plasma oxytocin and PGFM fluctuated between 5 and 20 pg/ml and 10 and 100 pg/ml respectively. Episodic pulses of plasma oxytocin (greater than 300 pg/ml) occurred on Days 15 and 16, whereas pulses of plasma PGFM (greater than 400 pg/ml) occurred on Days 19 and 20. There was little apparent correlation between episodic pulses of the two hormones. For 2 does that exhibited oestrus on Day 22, plasma progesterone concentrations declined to minimum values of 1.0-1.5 ng/ml by Day 20. One of these does showed very high levels of oxytocin secretion throughout the sampling period while the other showed an apparent paucity of oxytocin secretory periods. Two does hysterectomized on Day 13 of their second oestrous cycle failed to exhibit further oestrous cycles. Continual elevation of plasma progesterone concentrations (2-6 ng/ml) for an 8-month period indicated persistence of the corpus luteum after hysterectomy. It is concluded that luteolysis in fallow deer involves episodic secretion of both oxytocin and PGF-2 alpha.     

Secretion of gonadotrophins change during the luteal phase of the bovine oestrous cycle in the absence of corresponding changes in progesterone or 17β-oestradiol

The hypothesis in the present study was that changes in circulating luteinizing hormone (LH) and follicle stimulating hormone (FSH) would occur during the luteal phase of the oestrous cycle (Days 4–19; Day 0, day of behaviourial oestrus) that were not related to corresponding changes in concentrations of progesterone and 17β-oestradiol. The stage of the oestrous cycle of cows (n = 18) was synchronised to obtain cows that were on alternate days of the cycle. Blood samples were collected every other day at 15 min intervals for 12 h from all cows: Days 4, 6, 8, 10, 12, 14, 16, 18 (n = 9) and Days 5, 7, 9, 11, 13, 15, 17, 19 (n = 9). Concentrations of LH, FSH, 17β-oestradiol and progesterone were determined in these samples. Data were compared across days to determine when significant changes occurred in concentrations or patterns of secretion of the gonadotrophins and ovarian steroid hormones during the oestrous cycle. There were significant changes in mean concentrations of FSH in circulation between Days 6 and 12. The most striking changes in secretion of gonadotrophins that could not be explained by changes in gonadal steroids were the fluctuations in amplitude of LH pulses between Days 7 and 12. Amplitude of LH pulses increased between Days 7 and 11 and subsequently decreased between Days 11 and 12 of the oestrous cycle. Some changes in gonadotrophin secretion that occurred in the present study can be explained by fluctuations in concentrations of progesterone and 17β-oestradiol in circulation. Other changes cannot be explained by fluctuations in circulating concentrations of these steroids. We accept our hypothesis because the concomitant changes in mean concentration of FSH between Days 6 and 11 and amplitude of LH pulses between Days 7 and 12 of the bovine oestrous cycle cannot be explained by changes in circulating concentrations of progesterone and 17β-oestradiol.     

Growth and regression of ovarian follicles during the follicular phase of the oestrous cycle in heifers undergoing spontaneous and PGF-2 alpha-induced luteolysis

Ultrasonography was used to monitor the growth, ovulation and regression of individual ovarian follicles greater than or equal to 5 mm during the late luteal and follicular phases of the oestrous cycle in heifers treated with injections of PGF-2 alpha to induce luteolysis and in heifers undergoing spontaneous luteolysis. Six heifers were given a single injection of PGF-2 alpha between Day 12 and 15 of the oestrous cycle and their ovaries were examined daily by transrectal ultrasonography until ovulation occurred. Another group of 5 heifers was examined daily by ultrasound from Day 14 or 15 of the cycle through spontaneous luteolysis and ovulation. Blood samples were taken twice daily from this group and analysed for progesterone to determine when luteolysis occurred. All heifers were checked for oestrous behaviour twice daily. Mean diameters of ovulatory follicles on each of the 3 days before oestrus were not different between PGF-2 alpha-treated and untreated heifers. In both groups there was large variation among heifers in the sizes and growth rates of the ovulatory follicles. At 3 days before oestrus the diameters of ovulatory follicles were between 7.5 and 11 mm in PGF-2 alpha-treated heifers and between 6 and 11.5 mm in untreated heifers. Non-ovulatory follicles decreased in size during the 3 days before oestrus and the number of non-ovulatory follicles within the size ranges of ovulatory follicles decreased. The ovulatory follicle was not consistently the largest follicle on the ovaries until the day of oestrus but was always one of the 2 largest follicles during the 3 days before oestrus.(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of perioestrous suprabasal progesterone levels on cycle length,oestrous behaviour and ovulation in heifers

In order to induce suprabasal plasma concentrations of progesterone after luteolysis and to determine their effect on oestrous behaviour and ovulation, heifers subcutaneously received silicone implants containing 2.5 (n = 4), 5 (n = 4), 6 (n = 3), 7.5 (n = 3) or 10 (n = 4) g of progesterone, or an empty implant (controls, n = 5) between days 8 and 25 of the cycle (ovulation designated Day 0). Growth of dominant follicles and time of ovulation were determined by ultrasound, and signs of oestrus were recorded and scored. Blood was collected at 2–4 h intervals from Days 15 to 27 and assayed for progesterone concentration. In all heifers, plasma concentrations of progesterone sharply decreased during Days 16–18. Control heifers had their lowest progesterone levels on Days 20.5 and 21, standing oestrus on Day 19.5 ± 0.4 (mean ± SEM), and ovulated on Day 20.7 ± 0.4. A similar pattern was observed in heifers treated with 2.5 and 5 g progesterone. Heifers treated with 6, 7.5 and 10 g of progesterone showed an extended (P < 0.05) interovulatory interval. Onset of prooestrus and time of maximum expression of signs of oestrus were not significantly different from those in controls. However, there was an absence of standing oestrus in most of the cases, signs of oestrus lasted longer (P < 0.05) and were weaker in intensity when doses increased. In these groups, the lowest progesterone concentrations were attained shortly after implant removal. Some heifers treated with 6 and 7.5 g of progesterone had standing oestrus and post oestrous bleeding as seen in the controls but ovulation occurred from Days 24.5 to 27. When plasma progesterone concentrations were over 1 nmol 1⁻¹, disturbed oestrus and delayed ovulation occurred. The extended period of prooestrus and oestrus and delayed ovulation were similar to that described in cases of repeat breeding. It is suggested that suprabasal plasma concentrations of progesterone, after luteolysis, may lead to asynchrony between onset of oestrus and ovulation and consequently be a cause of repeat breeding in cattle.     

Pattern of follicular growth and resumption of ovarian activity in post-partum beef suckler cows

The ovaries of 18 post-partum beef suckler cows were examined daily, using ultrasound, from Day 5 post partum until a normal oestrous cycle was completed. Periods of growth and regression of medium-sized (5-9 mm) follicles were identified before one medium follicle became dominant (single large follicle greater than or equal to 10 mm). The mean (+/- s.e.m.) number of days from parturition to detection of the first post-partum dominant follicle was 10.2 +/- 0.5. The first post-partum dominant follicle ovulated in 2/18 (11%) cows. The interval from calving to first ovulation (mean +/- s.e.m. = 35.9 +/- 3.3 days) was characterized by the growth and regression of a variable number (mean = 3.2 +/- 0.2; range 1-6) of dominant follicles. The maximum diameter of the dominant follicle increased as the cows approached first ovulation (P less than 0.05). Behavioural oestrus was not detected in 16/18 (89%) cows at first ovulation. Following first ovulation, the length of the subsequent cycle was short (mean = 9.7 +/- 0.5 days; range 8-15 days) in 14/18 (78%) cows and was characterized by the development and ovulation of a single dominant follicle. During oestrous cycles of normal length (mean = 20.6 +/- 0.5 days; range 18-23 days) one (N = 2), two (N = 7) or three (N = 8) dominant follicles were identified. The growth rate, maximum diameter or persistence of non-ovulatory dominant follicles before first ovulation or during oestrous cycles were not different (P greater than 0.05). These data show that, in beef suckler cows, follicular development and formation of a dominant follicle occur early after parturition and the incidence of ovulation of the first dominant follicle is low. The number of dominant follicles that develop before first ovulation is variable; first ovulation is rarely associated with oestrus and short cycles are common after first ovulation. It is concluded that prolonged anoestrus in post-partum beef suckler cows is due to lack of ovulation of a dominant follicle rather than delayed development of dominant follicles.     

Episodic secretion of gonadotrophins and ovarian steroids in jugular and utero-ovarian vein plasma during the follicular phase of the oestrous cycle in gilts

Blood samples were collected simultaneously from the jugular and utero-ovarian veins of 13 gilts from Days 11 through 16 of the oestrous cycle. A luteolytic dose (10 mg) of PGF-2 alpha was given on Day 12 to facilitate the natural occurrence of luteolysis and standardize the associated decrease in concentrations of progesterone. The mean interval from PGF to oestrus was 5.5 +/- 0.7 days (mean oestrous cycle length = 17.5 +/- 0.7 days). Mean concentrations, pulse amplitudes and pulse frequencies of oestradiol and progesterone were greater (P less than 0.05) in the utero-ovarian than jugular vein. Secretory profiles of LH and FSH were similar (P greater than 0.05) in plasma collected simultaneously from both veins. Based on these data, temporal relationships among hormonal patterns of FSH and LH in the jugular vein and oestradiol and progesterone in the utero-ovarian vein were examined. Concentrations of progesterone declined (P less than 0.05) between Days 12 and 14, while all secretory variables for oestradiol increased (P less than 0.05) from Day 12 through 16 of the oestrous cycle. The pulsatile secretion of FSH remained relatively constant during the experiment. However, both pulse amplitude and mean concentration tended (P less than 0.2) to be lower on Day 16 compared with Day 12. The episodic secretion of LH shifted from a pattern characterized by high-amplitude, low-frequency pulses to one dominated by numerous pulses of diminishing magnitude between Days 13 and 14. From Days 14 to 16 of the oestrous cycle, 91% of all oestradiol pulses were temporally associated with gonadotrophin pulses composed of both FSH and LH episodes. However, pulses of oestradiol (52%) not associated with an episode of LH and/or FSH were observed on Days 12 and 13. These data demonstrate that during the follicular phase of the pig oestrous cycle substantial oestradiol production occurred coincident with luteolysis and before the shift in the episodic secretion of LH. The pool of follicles which ovulated was probably the source of this early increase in the secretion of oestradiol. Therefore, we propose that factors in addition to FSH and LH are involved in the initial selection of follicles destined to ovulate during the early stages of the follicular phase of the pig oestrous cycle. In contrast, high-frequency, low-amplitude pulses composed of LH and FSH were the predominant endocrine signal associated with oestradiol secretion during the second half of the oestrous cycle.(ABSTRACT TRUNCATED AT 400 WORDS)     

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.     

In situ analysis of the changes in expression of ovarian inhibin subunit mRNAs during follicle recruitment after ovulation in pigs

In situ hybridization was used on frozen tissue sections with digoxigenin-labelled antisense riboprobes to inhibin/activin alpha and beta(A) subunits to determine whether inhibin/activin subunit mRNA expression was associated with development of growing, steroidogenically active follicles during follicle recruitment after ovulation. Cell proliferation-associated nuclear antigen Ki-67 protein and cytochrome P450 aromatase expression in granulosa cells were determined immunohistochemically and used as markers for granulosa cell proliferation and steroidogenesis, respectively, on days 3, 5 and 7 after the onset of oestrus. The amounts of inhibin/activin alpha and beta(A) subunit mRNA and P450 aromatase protein were greater (102, 93, and 238%, respectively; P < 0.05) in medium than in small non-atretic follicles and were positively correlated with Ki-67 and with each other. Inhibin/activin alpha and beta(A) mRNA, P450 aromatase, and Ki-67 in granulosa cells were reduced by 66-83% (P < 0.001) in atretic follicles compared with non-atretic follicles. In addition, inhibin/activin alpha and beta(A) mRNA and P450 aromatase in small (1-2 mm) non-atretic follicles decreased (P < 0.05) between day 3 and day 7 independently of morphological or biochemical signs of atresia. The pattern of inhibin/activin subunit mRNA expression supports the notion that activin and inhibin have roles in growth and steroidogenesis in follicle recruitment during the early luteal phase of the oestrous cycle.     

Morphological and biochemical characteristics during ovarian follicular development in the pig

Ovaries were recovered from groups of naturally cyclic pigs (N = 5) on each of Days 16, 18, 20 and 21 of the oestrous cycle. Follicular diameter, follicular fluid volume and concentrations of oestradiol, testosterone and progesterone, and granulosa cell number were determined in all follicles greater than or equal to 2 mm in diameter (n = 511). In alternate follicles either granulosa cell aromatase activity and theca testosterone content or 125I-labelled hCG binding to granulosa and theca were determined. The mean total number of follicles recovered per animal decreased as the follicular phase progressed and a strong positive relationship (P less than 0.001) existed between follicular diameter and volume on all days. The number of granulosa cells recovered per follicle was variable, and not related to oestrogenic activity of the follicles. Mean follicular fluid oestradiol, testosterone and 125I-labelled hCG binding all increased until Day 20 and decreased on Day 21, whereas mean theca testosterone content, 125I-labelled hCG binding to theca tissue and aromatase were all maximal on Day 21. On Days 20 and 21 a subset of 14-16 large follicles was readily distinguishable from the remaining smaller, less oestrogenically active population in each animal. Yet, consistently within these subsets there was a difference in follicular diameter of approximately 2.0 mm and also a considerable range of biochemical development even among follicles of equal size. These results indicate asynchrony at the time of recruitment and selection among follicles destined to ovulate and suggest that heterogeneity continues into the immediate preovulatory period.     

Decreased superovulatory responses in heifers superovulated in the presence of a dominant follicle

Dairy heifers were superovulated in the presence (dominant group, N = 8) or absence (non-dominant group, N = 6) of a dominant follicle at the start of a a superovulatory treatment on Days 7-12 of the oestrous cycle (Day 0 = oestrus). Daily ultrasonographic observations of ovaries (recorded on videotape) starting on Day 3 were used to assess the presence or absence of a dominant follicle (diameter greater than 9 mm, in a growing phase or at a stable diameter for less than 4 days) and to monitor follicular development before and during treatment. The number of CL estimated by ultrasonography (7.1 +/- 1.8 vs 13.5 +/- 1.4) or by rectal palpation (6.9 +/- 2.0 vs 16.3 +/- 1.6) and mean progesterone concentrations (32.5 +/- 19 vs 80.7 +/- 16 ng/ml) after treatment were lower (P less than 0.01) in the dominant than in the non-dominant group. Based on number of CL, two populations of heifers were identified in the dominant group, i.e. those that had a high (dominant-high, N = 4; greater than 7 CL) or a low (dominant-low, N = 4; less than 7 CL) response to treatment. During treatment, the increases in number of follicles 7-10 mm and greater than 10 mm in diameter occurred sooner and were of higher magnitude in the non-dominant than in the dominant-high or dominant-low groups (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of mouse epidermal growth factor on plasma concentrations of FSH, LH and progesterone and on oestrus, ovulation and ovulation rate in merino ewes

The 24 h i.v. infusion of Merino ewes with 60 or 100 microgram mouse epidermal growth factor (EGF)/kg body weight on Days 4, 9 or 14 of the oestrous cycle decreased the strength of wool attachment and caused marked changes in subsequent reproductive performance. In ovaries removed 2 days after EGF treatment all follicles greater than or equal to 0.6 mm diameter were atretic. After 7 days either a normal pattern of atresia or no atresia was evident while after 12 days the pattern of follicular atresia was similar to that in controls. Irrespective of stage of cycle EGF caused dose-dependent increases in plasma FSH concentrations that persisted for up to 14 days. Changes in plasma LH concentrations were generally similar after infusion on Days 4 and 14, but were smaller and shorter-lived after infusion on Day 9. Irrespective of dose, the infusion of EGF on Days 4 and 14 caused immediate luteolysis then the formation of a luteinized follicle in many ewes. Most ewes treated on Day 4 returned to oestrus between Days 17 and 21 with the same ovulation rate (1.3) as the controls. Of those infused on Day 14 oestrus occurred about a cycle length later than expected and their ovulation rate then (1.9) was also similar to that of the controls (1.7). Luteal function was not affected in ewes infused on Day 9, and most returned to oestrus between Days 17 and 20 with an ovulation rate of 3.2. Fertile rams were not placed with the ewes until after the differences in ovulation rate had been observed. Mating occurred generally 2-4 weeks after treatment, and there were no differences between EGF-treated and control ewes in fertility or fecundity. The results are interpreted as indicating that mouse EGF induces ovarian follicular atresia but has differential effects on luteal function according to the stage of the oestrous cycle at which it is given. As a consequence of these two effects, which lead to differential changes in gonadotrophin secretion, ovarian function may be temporarily impaired, little affected or improved.     

The mature mid-cycle follicle in the mare.

Oestrogen and progesterone concentrations in blood and follicular fluid and blood levels of LH were determined in 426 mares at different stages of the oestrous cycle. Mature follicles occur at all stages of the cycle; they ovulate readily in early metoestrus, occasionally in late metoestrus and very rarely in dioestrus. Maturation of a mid-cycle follicle is associated with intermediate levels of LH, which are less than those found during oestrus. This lower level of LH together with a high level of progesterone are probably responsible for the failure of ovulation and regression of most of the mid-cycle mature follicles found in the mare.     

Role of luteinizing hormone in follicle deviation based on manipulating progesterone concentrations in mares

The effects of several doses of progesterone on FSH and LH concentrations were used to study the role of the gonadotropins on deviation in growth rates of the two largest follicles during the establishment of follicle dominance. Progesterone was given to pony mares at a daily dose rate of 0 mg (controls), 30 mg (low dose), 100 mg (intermediate dose), and 300 mg (high dose). All follicles > or = 6 mm were ablated at Day 10 (Day 0 = ovulation) to initiate a new follicular wave; prostaglandin F(2alpha) was given to induce luteolysis, and progesterone was given from Days 10 to 24. The low dose did not significantly alter any of the ovarian or gonadotropin end points. The high dose reduced (P < 0.05) the ablation-induced FSH concentrations on Day 11. Maximum diameter of the largest follicle (17.2 +/- 0.6 mm) and the second-largest follicle (15.5 +/- 0.9 mm) in the high-dose group was less (P < 0.04) than the diameter of the second-largest follicle in the controls (20.0 +/- 1.0 mm) at the beginning of deviation (Day 16.7 +/- 0.4). Thus, the growth of the two largest follicles was reduced by the high dose, presumably through depression of FSH, so that the follicles did not attain a diameter characteristic of deviation in the controls. The intermediate dose did not affect FSH concentrations. However, the LH concentrations increased in the control, low, and intermediate groups, but then decreased (P < 0.05) in the intermediate group to pretreatment levels. The LH decrease in the intermediate group occurred 2 days before deviation in the controls. The maximum diameter of the largest follicle was less (P < 0.0001) in the intermediate group (27.3 +/- 1.8 mm) than in the controls (38.9 +/- 1.5 mm), but the maximum diameter of the second-largest follicle was not different between the two groups (19.0 +/- 1.1 vs. 20.3 +/- 1.0 mm). Thus, the onset of deviation, as assessed by the second-largest follicle, was not delayed by the decrease in LH. Diameter of the largest follicle by Day 18 in the intermediate group (23.1 +/- 1.6 mm) was less (P < 0.05) than in the controls (28.0 +/- 1.0 mm). These results suggest that circulating LH was not involved in the initiation of dominance (inhibition of other follicles by the largest follicle) but was required for the continued growth of the largest follicle after or concurrently with its initial expression of dominance.     

Effect of treatment with progesterone and oestradiol when starting treatment with an intravaginal progesterone releasing insert on ovarian follicular development and hormonal concentrations in Holstein cows

Ovarian follicular development and concentrations of gonadotrophin and steroid hormones were studied in non-lactating Holstein cows following administration of progesterone (P(4)) or oestradiol benzoate (ODB) at the start of treatment with an intravaginal progesterone releasing insert (IVP(4)) in a 2 by 2 factorial experiment. Cows were treated at random stages of the oestrous cycle with an IVP(4) device (Day 0) and either no other treatment (n=8), 200 mg of P(4) IM (n=9), 2.0 mg of ODB IM (n=8) or both P(4) and ODB (n=9). Seven days later devices were removed and PGF(2alpha) was administered. Twenty-four hours later 1.0mg of ODB was administered IM. Oestrus was detected in 97.1% and ovulation in 64.7% (effect of treatment, P=0.41) of cows within 96 h of removing inserts. In the cows that ovulated, day of emergence of the ovulatory follicle was delayed (P<0.01) and more precise (P<0.05) in cows treated with ODB compared to the cows treated with P(4). Interval from wave emergence to ovulation and the diameter of the ovulatory follicle was less (P<0.05) in cows treated with ODB compared to cows treated with P(4). Combined treatment with P(4) and ODB at the time of starting treatment with an IVP(4) device did not significantly change the pattern of ovarian follicular development compared to treatment with ODB alone. Concentrations of LH and FSH in plasma were less in cows treated with ODB between Days 0 and 4 (P<0.05) while treatment with P(4) increased concentrations of FSH in plasma between Days 0 and 4 (P<0.05). When anovulatory cows were compared to ovulatory cows, diameters of follicles (P<0.001) and growth rate of follicles (P<0.01) were less in anovulatory cows between Days 7 and 9, while concentrations of FSH in plasma were greater (P<0.01), concentrations of LH similar (P>0.90) and concentrations of oestradiol were less (P=0.01) in the anovulatory cows between Days 4 and 10. Our findings support a hypothesis that ovarian follicular development following administration of P(4) or ODB at the start of treatment with an IVP(4) device differs. Anovulatory oestrus may have been associated with reduced maturity and/or later emergence of ovarian follicles.     

Oestrous and ovarian responses to exogenous progesterone and oestrogen in prepubertal ewe lambs in relation to the fecundity of their dams

Oestrus was consistently induced by 20, 40 or 80 microgram oestradiol benzoate in progesterone-primed lambs aged 23 weeks and 17 weeks respectively in two experiments. The duration of oestrus increased linearly with increasing log dose of oestrogen, and was negatively correlated with the time of its onset. In the absence of progesterone there was a reduced incidence, later onset and shorter duration of oestrus. Progesterone alone did not induce oestrus. In lambs treated during January (Exp. 2), a later date of injection of oestrogen was associated with earlier onset and longer duration of oestrus. The induced oestrus was anovulatory. Oestrogen reduced the proportion of lambs with follicles greater than or equal to 3 mm in diameter whilst progesterone had no effect. Lambs which were the progeny of low- and high-fecundity dams did not differ in their oestrous or ovarian responses. Correlations between the dam's lamb production index and the time to onset and duration of the induced oestrus were also not significant.     

Regulation of circulating gonadotropins by the negative effects of ovarian hormones in mares

The functional and temporal relationships between circulating gonadotropins and ovarian hormones in mares during Days 7-27 (ovulation = Day 0) was studied using control, follicle ablation, and ovariectomy groups (n = 6 mares/group). In the follicle-ablation group, all follicles > or = 6 mm were ablated on Day 7, and every 2 days thereafter, newly emerging follicles were also ablated. Estradiol concentrations decreased (P < 0.01) similarly in the controls and the follicle-ablation group between Days 7 and 11 and by Day 15 began to increase in the controls and continued to decrease in the follicle-ablation group. Concentrations of progesterone were not affected by follicle ablation, but diameter of the corpus luteum was greater (P < 0.05) by Day 21 in the follicle-ablation group; these results indicated that the follicles were involved in morphologic luteolysis, but not in functional luteolysis. Concentrations of LH were higher (P < 0.05) on Days 15 and 16 in the follicle-ablation group than in the controls, indicating an initial negative effect of follicles on LH. Immunoreactive inhibin and estradiol decreased (P < 0.0001) and FSH and LH increased (P < 0.05) within 1 or 2 days after ovariectomy; these changes occurred more slowly in the follicle-ablation group. The maximum value for an FSH surge in each control mare was below the lower 95% confidence limit in the ovariectomy group. Maximum concentration for the periovulatory LH surge in the controls was not different from the mean maximum LH concentrations in the ovariectomy group. Our interpretation is that the gonadotropin surges resulted from changes in the magnitude of the negative effects of ovarian hormones on the positive effects of extraovarian control. There was no indication of a positive ovarian effect on either FSH or LH.