Factors that affect ovarian follicular dynamics in cattle.

Studies of ovarian follicular dynamics in cattle may lead to methods for improving fertility, for synchronizing estrus with more precision, and for enhancing superovulatory responses. Within an estrous cycle, two or three large (> 10 mm) follicles develop during consecutive waves of follicular growth. The last wave provides the ovulatory follicle, whereas preceding wave(s) provide follicles that undergo atresia. The life span of large follicles seems to depend on the pulsatile secretion of LH; decreased frequency of LH pulses results in atresia of large follicles. Aromatase activity in the walls of the largest follicles is greatest during the first 8 d of the estrous cycle and decreases by d 12. Steroidogenesis of the largest and second-largest ovarian follicles differs on d 5, 8, and 12 of the estrous cycle. Follicular dynamics are altered by negative energy balance and lactation. The number of large follicles and concentration of estradiol during the preovulatory period differs between postpartum lactating and nonlactating cows. Dietary fats stimulate follicular growth when they are fed to increase energy balance. Administration of bovine somatotropin decreases energy balance and has a differential effect on ovarian follicular responses; growth of the largest follicle does not change, but growth of the second-largest follicle is stimulated by somatotropin. Studies of follicular dynamics in lactating cows demonstrate changes in ovarian function associated with energy balance that may be related to inefficient reproductive performance of cows producing high yields of milk.     

Biochemical analysis of bovine follicular fluid: albumin, total protein, lysosomal enzymes, ions, steroids and ascorbic acid content in relation to follicular size, rank, atresia classification and day of estrous cycle

To investigate some biochemical changes during bovine follicle development, ovaries were obtained from cyclic heifers (7 to 11 heifers/d on each day of the 21-d estrous cycle; N = 152). Follicular fluid from the two largest follicles from both ovaries and a pool from small follicles (N = 30/cow) were collected from each animal and analyzed for ionic, enzymatic and endocrine changes in relation to day of the estrous cycle, follicle size, rank and atretic or growing status. Follicular fluid alkaline phosphatase activity and ascorbate concentrations were highest in all follicular sizes during the earlier portion of the estrous cycle (d 1 to 12; P less than .05), then decreased to the lowest levels (d 13 to 21). As follicular size (diameter) increased lactate dehydrogenase (LDH), acid and alkaline phosphatase activity was reduced in follicular fluid (P less than .05). Alkaline phosphatase and LDH activity tended to be increased in atretic follicles (P less than .10), and was correlated with increased progesterone and androgen concentrations of follicular fluid (r = .4, P less than .05). Both albumin and total protein concentrations decreased as follicular diameter increased (P less than .05). Sodium concentrations in follicular fluid were greater in growing-antral than atretic follicles, and increased with follicular enlargement (P less than .05). Follicular potassium concentrations increased as the estrous cycle progressed (P less than .05), and tended to be elevated in atretic follicles (nonsignificant). Both Ca and Mg concentrations increased with follicular enlargement (P less than .05). Dehydroepiandrosterone and testosterone were the predominant androgens in follicular fluid (androstenedione, the lowest concentration); their concentration decreased with follicle development (P less than .05), but were quite variable. Estradiol was increased in growing follicles (P less than .01). Estrone and estradiol concentrations increased as ovulation approached, particularly in small follicles (less than or equal to 4 mm diameter). Changes of biochemical components found in follicular fluid that relate to the growth and atresia process may provide a more sensitive and accurate method to classify follicle status, and thus aid in understanding the complexity of events associated with maturation of the bovine follicle and oocyte.     

Ovarian follicular development in cattle selected for twin ovulations and births

Comparisons of numbers of antral ovarian follicles and corpora lutea (CL), of blood hormone concentrations, and of follicular fluid steroid concentrations and IGFBP activity were conducted between cows selected (twinner) and unselected (control) for twin births to elucidate genetic differences in the regulation of ovarian follicular development. Ovarian follicular development was synchronized among cows by a single i.m. injection of PGF2alpha on d 18 of the estrous cycle; six cows per population were slaughtered at 0, 24, 48, and 72 h after PGF2alpha. Jugular vein blood was collected from each animal at PGF2alpha injection and at 24-h intervals until slaughter. Ovaries of twinner cows contained more small (< or = 5 mm in diameter, P < 0.05), medium (5.1 to 9.9 mm, P < 0.05), and large (> or = 10.0 mm, P < 0.01) follicles and more (P < 0.01) CL than ovaries of controls. Follicular fluid concentrations of estradiol, androstenedione, testosterone, and progesterone reflected the stage of follicular development and were similar for twinner and control follicles at the same stage. Earlier initiation of follicular development and/or selection of twin-dominant follicles in some twinner cows resulted in greater concentrations of estradiol in plasma at 0, 24, and 48 h and of estradiol, androstenedione, and testosterone in follicular fluid of large follicles at 0 h after PGF2alpha for twinner vs. control cows (follicular status x time x population, P < 0.01). Binding activities of IGFBP-5 and -4 were absent or reduced (P < 0.01) in follicular fluid of developing medium and large estro-gen-active (estradiol:progesterone ratio > 1) follicles but increased with atresia. Only preovulatory Graafian follicles lacked IGFBP-2 binding, suggesting a possible role for IGFBP-2 in selection of the dominant follicle. Concentrations of IGF-I were twofold greater (P < 0.01), but GH (P = 0.10) and cholesterol (P < 0.05) were less in blood of twinners. Three generations of selection of cattle for twin ovulations and births enhanced ovarian follicular development as manifested by increased numbers of follicles within a follicular wave and subsequent selection of twin dominant follicles. Because gonadotropin secretion and ovarian steroidogenesis were similar for control and twinner cattle, enhanced follicular development in twinners may result from decreased inhibition by the dominant follicle(s), increased ovarian sensitivity to gonadotropins, and/or increased intragonadal stimulation, possibly by increased IGF-I.     

Ovarian follicular dynamics and hormones throughout the estrous cycle in Thai native (Bos indicus) heifers

Relatively few studies have been reported regarding the reproductive physiology of female Thai native cattle. Therefore, the objective of the present study was to evaluate the follicular dynamics and concentrations of follicle stimulating hormone (FSH), estradiol (E2) and progesterone (P4) during the estrous cycle in Thai native heifers (TNH) and to compare obtained results with those of European and Indian cattle breeds previously reported. For the detection of estrus, ovaries of all 20 heifers were examined twice daily (12 h intervals) by ultrasonography for three consecutive estrous cycles. From data of 60 estrous cycles (n = 60 estrous cycles from 20 heifers), it was found that 14 (70%) and 6 heifers (30%) had two (42 estrous cycles collected from 14 heifers) and three follicular waves (18 estrous cycles collected from 6 heifers), respectively. The days when estrus was detected, interovulatory intervals, life‐spans of corpus lutea (CL), and days for growing and regression of CLs were shorter in the two follicular waves than those in the three follicular waves (P < 0.05). In both two and thre follicular waves, larger maximum diameters and higher growth rates of the dominant follicle (DF) in an ovulatory wave were observed than those of the preceding waves without ovulation (P < 0.05). There was a progressive increase in follicular size and FSH and E2 production during follicular growth in each follicular wave. In addition, the FSH and E2 peak concentrations during the ovulatory wave were higher than those of the anovulation waves (P < 0.05). Moreover, although the ovarian follicular dynamic patterns in Thai native heifers were similar to those previously reported for European and Indian cattle breeds, the diameter of the largest preovulatory follicle (OF), subordinate follicles (SF) and CLs were smaller than those in European and Indian cattle breeds. In conclusion, when compared with European and some breeds of Indian cattle, the length of interovulatory intervals was shorter, and the sizes of dominant SF and CLs were smaller in Thai native heifers.     

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.     

Endocrine changes and ultrasonography of ovaries in suckled beef cows during resumption of postpartum estrous cycles

Changes in follicular and luteal structures were assessed and concentrations of estradiol and progesterone were measured in 13 Hereford X Angus suckled beef cows during resumption of estrous cycles. Transrectal ultrasonography was used to monitor follicular size, ovulation, and formation and regression of the corpus luteum (CL). The interval from parturition to first postpartum ovulation (FO) was 82 +/- 4.7 d. Serum progesterone remained low before FO. One cow exhibited standing estrus, two cows showed other signs of estrus, and 10 displayed no signs of behavioral estrus preceding FO. All cows exhibited standing estrus before the second postpartum ovulation (SO). All cows had a short luteal phase after FO, with an average interval of 8.5 +/- .2 d between FO and SO. Concentrations of estradiol in serum during the 8 d preceding ovulation were similar before FO and SO. Maximal diameter of the preovulatory follicle was similar before FO and SO. However, the ovulatory follicle was larger in diameter at 2 d (P = .02) and 3 to 8 d (P less than .005) before FO than before SO. The time from detection until ovulation was less (P = .005) for the ovulatory follicle preceding SO than for the follicle associated with FO (8.5 vs 10.2 d, respectively, SE = .4). The second-largest follicle was larger (P less than .005) in diameter during the 8 d preceding the FO than before the SO. The difference in size between the ovulatory follicle and the second-largest follicle on the day before ovulation was greater (P less than .005) preceding SO than preceding FO (8.7 vs 6.6 mm, respectively, SE = .4).(ABSTRACT TRUNCATED AT 250 WORDS)     

Histological Characteristics and Steroid Concentration of Ovarian Follicles at Different Stages of Development in Pregnant and Non-pregnant Dairy Cows

The aim of the study was to investigate the histological characteristics and steroid concentrations in follicular fluid of different populations of follicles at different stages of development, during pregnancy and the oestrous cycle in cows. Follicles from ovaries collected at a slaughterhouse were allocated into three size categories (small, 2–5.9 mm; medium, 6–13.9 mm; and large, 14–20 mm) in pregnant and non-pregnant cows. Slices were stained with HE and PAS for histological analysis. Follicular fluid was pooled according to size and pregnancy status and estradiol, testosterone and progesterone concentrations in follicular fluid were determined by RIA. Characteristics of healthy follicles did not differ, regardless of follicle size or pregnancy status. Total histological atresia was significantly higher in pregnant cows than in non-pregnant cows (p < 0.05). Estradiol increased and testosterone decreased significantly, while follicles increased in size, in both non-pregnant and pregnant cows (p < 0.05). Nonpregnant cows had the highest estradiol values in follicles of all sizes. Medium and large follicles from pregnant cows showed the lowest testosterone concentration (p < 0.05). Progesterone levels increased with follicle size only in non-pregnant animals. In large follicles, progesterone concentration was significantly higher in non-pregnant cows than in pregnant cows (p < 0.05). Considering steroid concentration and histological findings, most large follicles might be atretic during pregnancy in cattle.     

Dynamics of ovarian follicular development in cattle following hysterectomy and during early pregnancy

Three experiments were conducted to evaluate ovarian follicular dynamics and functional activity during pregnancy in cattle. In 11 pregnant Charolais cows of Experiment I, size of largest follicle, number of follicles and accumulated follicle size were reduced by day 27 of pregnancy on the ovary bearing the corpus luteum (CL) but not on the non-CL bearing ovary. In experiment II, local attenuation of ovarian follicular development on the CL bearing ovary of seven pregnant heifers was evident compared to the contralateral ovary without the CL. However, in four hysterectomized heifers, follicular development was sustained on both the CL- and non-CL bearing ovaries when CL maintenance was achieved without presence of the uterus or conceptus. In Experiment III, steroidogenic characteristics of the largest and second largest follicles at 17 d postestrus were evaluated for seven pregnant and six cyclic cattle. Follicle by physiological status interactions were detected for both aromatase activity of the follicle and follicular fluid concentrations of estradiol and progesterone. In cyclic cows, the largest follicle had appreciably more aromatase activity than did the second largest follicle; whereas, aromatase activity of the largest follicle from pregnant cows was less than that of cyclic cows. However, in pregnant cows the second largest follicle became the estrogen-active follicle, and this follicle occurred with a higher frequency on the ovary contralateral to the CL-bearing ovary. These changes in aromatase activity were reflected by parallel changes in estrogen concentrations of follicular fluid. The higher progesterone concentration in follicular fluid of the largest follicle in pregnant cows provided further confirmation of their atretic status. In conclusion, during early pregnancy the conceptus and/or uterus ipsilateral to the conceptus appear to secrete compounds which alter local follicular steroidogenic activity and attenuate subsequent follicular growth between 17 to 34 d of pregnancy on the CL-bearing ovary. This local mechanism acting within the ovary may contribute to the antiluteolytic effects of early pregnancy in cattle.     

Follicular development and maturation in gilts selected for an index of high ovulation rate and high prenatal survival

Seventy-one 10th-generation gilts from White Line-1 (WL-1 = randomly selected control line) and White Line-2 (WL-2 = selected for an index of ovulation rate and prenatal survival rate) were used to compare the pattern of follicular development and atresia during the follicular phase of the estrous cycle. Gilts were treated with PGF(2alpha)on d 13 of the estrous cycle (d 0 of induced follicular development) to induce luteolysis and assigned randomly within line and sire for ovary recovery on d 0, 2, 3, 4, 5, and the day after estrus. Ovaries were evaluated for numbers of corpora albicantia and small (2 to 2.9 mm), medium (M1 = 3 to 4.9 mm; M2 = 5 to 6.9 mm), and large (>or=7 mm) follicles. The concentration of estradiol-17beta in follicular fluid was used to classify individual M2 and large follicles as estrogen-active (>or=100 ng of estradiol-17beta/mL) or inactive (<100 ng of estradiol-17beta/mL). The WL-2 gilts had a greater ovulation rate than WL-1 gilts at their pre-treatment estrus (20.4 vs. 13.8 corpora albicantia; P < 0.001). The small and M1 follicle populations decreased rapidly in both lines over time (P < 0.001). The M2 follicle population increased in both lines between d 0 to 4 and then decreased. Mean estradiol concentration of M2 follicles increased in both genetic lines over time (P < 0.02). All large follicles were estrogen-active in both lines; the number of large follicles increased with day (P < 0.001) and was similar in both lines. The number of estrogen-active M2 follicles was similar in both lines, increasing to d 3 and 4 and then decreasing (P < 0.01) thereafter. However, the total number of estrogen-active follicles (sum of estrogen-active M2 and large follicles) was greater in WL-2 than in WL-1 gilts (P < 0.04), increasing to the ovulatory potential by d 3 in WL-1 gilts, but continuing to increase through d 4 in WL-2 gilts. Selection of an additional six ovulatory follicles from the estrogen-active M2 follicle pool after d 5 was required in both lines to achieve the projected ovulation rate, and after estrus, the number of large follicles remained insufficient to attain the ovulatory potential of each line.     

Incidence, Endocrinology, Vascularity, and Morphology of Hemorrhagic Anovulatory Follicles in Mares

The incidence of hemorrhagic anovulatory follicles (HAFs) is approximately 5% and 20% of estrous cycles during the early and late ovulatory season, respectively. The structures are more common in old mares (eg, >20 years), tend to occur repeatedly in individuals, and occur most frequently during the late follicular phase. In a recent study, the day of ovulation in controls and the first day of HAF formation, as indicated by cloudiness of follicular fluid, were defined as day 0. On day -1, future ovulating and HAF groups did not differ in follicle diameter or in the frequency of discrete gray-scale ultrasonic indicators of impending ovulation; however, in future HAFs, a greater percentage of the circumference of the follicle exhibited color-Doppler signals of blood flow. No differences were found between the two groups in systemic concentrations of progesterone, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) on days -4 to 2, but estradiol was elevated in the HAF group on day -3. The wall of the HAFs developed well-vascularized luteal tissue as indicated by echotexture and color Doppler signals and by the production of near normal levels of progesterone. In conclusion, HAFs formed from viable preovulatory follicles that did not differ from ovulatory follicles in diameter or gray-scale echotexture. Estradiol concentrations were elevated a few days before the failure of ovulation, and the wall of the follicle was more extensively vascularized on day -1.     

Follicular Fluid Prolactin and the Periovulatory Prolactin Surge in the Mare

Prolactin may play multiple roles in equine reproduction. Prolactin appears to be associated with seasonal reproduction, and fluctuating prolactin levels during the estrous cycle suggest that it may play a role in estrous cyclicity as well. The purpose of this research was to investigate the activity of prolactin during the follicular phase of the estrous cycle. In experiment 1, prolactin concentrations were determined from plasma samples collected at least every other day throughout the estrous cycle. Periovulatory (ovulation ± 1 day) prolactin concentrations were compared with concentrations during early diestrus (days 2−10 postovulation). In experiment 2, prolactin concentrations were measured in follicular fluid collected from 74 follicles of various sizes. Follicles were grouped into small (≤20 mm), medium (21−35 mm), and large (>35 mm) size categories. Prolactin concentrations increased during the periovulatory period in cycling mares. This periovulatory surge was superimposed on baseline prolactin concentrations that varied with season. Prolactin was present in significant quantities in the follicular fluid. Follicular fluid prolactin concentrations were lowest in small follicles and increased in medium and large follicles. Concentrations did not differ between medium and large follicles. Follicular fluid prolactin concentrations were lower in autumnal follicles compared with summer follicles of comparable size. It is possible that the short-term surge in circulating prolactin around ovulation could be linked to the significant levels of prolactin in follicular fluid. Ovulation releases a relatively large volume of fluid into the peritoneum. The prolactin in this fluid could be a contributor to the periovulatory prolactin surge.     

Follicular development,oocyte viability and recovery in relation to follicular steroids,prolactin and glycosaminoglycans throughout the estrous period in superovulated heifers with a normal LH surge,no detectable LH surge,and progestin inhibition of LH surge

Estrous cycles of heifers (n = 137) were synchronized with prostaglandin (PGF_2α) and follicular development stimulated with follicle stimulating hormone. Twenty-eight animals were administered Norgestomet implants 12 hr prior to the initial PGF2α injection to suppress the LH surge that initiates ovulation. Animals were ovariectomized every 12 hr after the initial PGF2α (7–9/time, 12–108 hr and at 192 and 240 hr post PGF2α) and divided into three treatment groups to consist of: 1) animals exhibiting a normal luteinizing hormone (LH) surge (n = 86), 2) animals in which no LH surge was detected (n = 23), and 3) suppression of the LH surge via Norgestomet implants (72–108 hr, n = 28). Follicular diameter was measured and follicular fluid was collected for analysis of prolactin, estradiol, progesterone and glycosaminoglycan concentrations. Progesterone concentrations were increased in animals exhibiting an LH surge as compared to animals in which no LH surge was detected; primarily in large follicles (> 8 mm diameter) after the LH surge. Animals not exhibiting an LH surge also had increased follicular progesterone concentrations compared to Norgestomet-implanted animals (242.3 ± 36.3 vs 86.7 ± 6.4 ng/ml, respectively, P < .01), indicating some LH stimulation. Follicular estradiol in animals exhibiting an LH surge increased up to the time of LH surge detection and then declined whereas animals with no LH surge detected had follicular estradiol concentrations that declined after the PGF_2α injection. No differences were noted between those that did not exhibit an LH surge or in which the LH surge was suppressed with Norgestomet in relation to follicular estradiol concentrations. Follicular estradiol concentrations increased with follicular size in all treatment groups (P < .01). Follicular concentrations of prolactin were increased in small follicles (P < .05; ≤ 4 mm diameter) and follicular prolactin increased from 12 to 36 hr post PGF2α injection, then declined after the LH surge. Follicular glycosaminoglycan concentrations decreased with increases in follicular size (P < .01) and were higher in animals that did not exhibit an LH surge (P < .01). No differences in follicular glycosaminoglycans were noted between Norgestomet-implanted animals and those not exhibiting an LH surge. In the animals representing days 4 and 6 of the subsequent estrous cycle (192 and 240 hr post PGF2α), numbers of small-sized follicles were increased. Follicular progesterone and estradiol concentrations were related to atretic large follicles unovulated from the prior estrus and a wave of growth in small and medium follicles. Follicular prolactin and glycosaminoglycans increased with time of the new estrous cycle and were increased in smaller follicles (P < .01). Suppression of LH with progestin implants (Norgestomet) may relate to early effects of progesterone, which may not be totally eliminated at target tissues and subsequently alters the LH surge, steroidogenesis of the follicle, and ovulation. Oocytes were predominantly found in the follicular fluid from animals in which an LH surge was detected and in the buffer wash of follicles in which no LH surge was detected. Oocyte viability was higher in animals exhibiting an LH surge (75% viable) whereas the oocytes of Norgestomet-implanted animals were 75% degenerate.     

Follicle-stimulating hormone isoforms and plasma concentrations of estradiol and inhibin A in dairy cows with ovulatory and non-ovulatory follicles during the first postpartum follicle wave

Following parturition, all cows display a wave of ovarian follicular growth, but a large proportion fail to generate a preovulatory rise in estradiol, and hence fail to ovulate. Follicle-stimulating hormone (FSH) exists as multiple isoforms in the circulation depending on the type and extent of glycosylation, and this has pronounced effects on its biological properties. This study examined differences in plasma FSH, estradiol, and inhibin A concentrations, and the distribution of FSH isoforms in cows with ovulatory or atretic dominant follicles during the first postpartum follicle wave. Plasma FSH isoform distribution was examined in both groups during the period of final development of the dominant follicle by liquid phase isoelectric focusing. Cows with an ovulatory follicle had higher circulating estradiol and inhibin A concentrations, and lower plasma FSH concentrations. The distribution of FSH isoforms displayed a marked shift toward the less acidic isoforms in cows with ovulatory follicles. A higher proportion of the FSH isoforms had a pI>5.0 in cows with ovulatory follicles compared to those with atretic follicles. In addition, cows with ovulatory follicles had greater dry matter intake, superior energy balance, elevated circulating concentrations of insulin and insulin-like growth factor-I, and lower plasma nonesterified fatty acids. The shift in FSH isoforms toward a greater abundance of the less acidic isoforms appears to be a key component in determining the capability for producing a preovulatory rise in estradiol, and this shift in FSH isoforms was associated with more favorable bioenergetic and metabolic status.     

Follicular growth and estradiol concentrations in foaling and nonparturient mares

Plasma estradiol concentration and follicular development were evaluated daily during the first postpartum estrus and the subsequent cycle of five foaling mares. For comparison, one estrous cycle was monitored in the same fashion for five nonparturient mares. The first postpartum estrous cycles were shorter but similar to non-pregnant cycles in ovarian steroid production and follicular activity. However, estradiol production from postpartum follicles was lower per mm follicular diameter than from follicles in nonpregnant cycles (p<0.05).     

Pregnancy-associated plasma protein-A and insulin-like growth factor binding protein mRNAs in granulosa cells of dominant and subordinate follicles of preovulatory cattle

To determine if (1) levels of pregnancy-associated plasma protein-A (PAPP-A) mRNA and insulin-like growth factor binding protein (IGFBP) (-2, -3, -4 and -5) mRNAs differ between the dominant and subordinate follicles during the follicular phase of an estrous cycle, and (2) these differences are associated with differences in follicular fluid (FFL) concentrations of steroids (estradiol, androstenedione, and progesterone), total and free IGF-I, or IGFBPs, estrous cycles of non-lactating Holstein dairy cows (n = 16) were synchronized with two injections of prostaglandin (PGF2 alpha) 11 days apart. Granulosa cells and FFL were collected either 24 h or 48 h after the second injection of PGF2 alpha. FFL from dominant follicles had lower concentrations of progesterone (P < 0.08) and higher concentrations of estradiol (P < 0.05), androstenedione (P < 0.0001), estradiol:progesterone ratio (P < 0.0001), free IGF-I (P < 0.0001), and calculated percentage free IGF-I (P < 0.01) than large subordinate follicles. Levels of IGFBP-2, -4, and -5 in FFL were 3.0- (P < 0.05), 2.4- (P < 0.06), and 3.4-fold (P < 0.05) greater, respectively, in subordinate than in dominant follicles. IGFBP-3, IGFBP-4 and PAPP-A mRNA expression and IGF-II concentration did not differ (P > 0.10) between dominant or subordinate follicles. Levels of IGFBP-2 and -5 mRNA were severalfold greater (P < 0.05) in subordinate than dominant follicles. IGFBP-5 mRNA in granulosa cells decreased (P < 0.05) 62% to 92%, between 24h and 48 h post-PGF2 alpha. We conclude that decreased levels of IGFBP-2 and -5 mRNA in granulosa cells may contribute to the decrease in FFL IGFBP-2 and -5 protein levels of preovulatory dominant follicles, and that changes in granulosa cell IGFBP-3 and -4 mRNA and PAPP-A mRNA levels do not occur during final preovulatory follicular development in cattle.     

Acid and alkaline phosphatase in bovine antral follicles

Acid and alkaline phosphatases were measured in the follicular fluid of 766 individual follicles from 96 cows. Follicles were obtained by bilateral ovariectomy or at slaughter from animals at various stages of the estrous cycle and pregnancy. Mean follicle size varied with the physiological state of the cow (P less than .0001). Acid phosphatase activity (U/microliters) varied inversely with follicle size (P less than .001) but not with stage of the estrous cycle or gestation. Total acid phosphatase activity per follicle increased with follicle size (P less than .05). Neither acid phosphatase nor alkaline phosphatase concentration was associated with atresia. Alkaline phosphatase activity (U/microliters) was greater in the smallest follicles (less than 50 microliters) than in other size groups (P less than .0001). Alkaline phosphatase activity (U/microliters) was greater (P less than .05) during the preovulatory phase of the estrous cycle than during other phases. A high concentration of follicular fluid phosphatases cannot be used as a marker for atresia but is characteristic of healthy small antral follicles.     

Effect of short-term calf removal at three stages of a follicular wave on fate of a dominant follicle in postpartum beef cows.

The hypothesis that ovulation in response to short-term (48 h) calf removal (CR) is dependent on the developmental stage of the dominant follicle was tested in two studies. The objective of Exp. 1 was to characterize the fate of a dominant follicle following 48-h CR on d 2, 4, or 8 of a postpartum follicular wave. Ovaries of 61 beef cows were examined daily by transrectal ultrasonography starting at d 20 to 21 postpartum. Treatments were no CR (n = 14) and CR on d 2 (n = 12), 4 (n = 16), or 8 (n = 10) of first detected follicular wave. Percentage of cows that ovulated a dominant follicle following treatment was not different among groups (P = 0.62). Maximum size of dominant follicles was larger in cows that ovulated (P = 0.002) than in cows that did not ovulate. The objectives of Exp. 2 were 1) to determine whether a follicular wave could be synchronized in anestrous cows following injection of 1 mg of estradiol benzoate (EB) and 200 mg of progesterone (P4; EB + P4); 2) to characterize the fate of dominant follicles following 48-h CR at three stages of a synchronized follicular wave; and 3) to determine whether estrous cycles of normal length followed ovulation in cows pretreated with EB + P4. Ovaries of 50 anestrous beef cows were examined daily as in Exp. 1. Treatments were sesame oil (SO) injected (i.m.) on d 25 postpartum and no CR (n = 9); EB + P4 and no CR (n = 9); EB + P4 and CR on 6 (n = 12), 8 (n = 9), or 12 (n = 11) d after injection. The EB and P4 injections were given on d 25 postpartum. Variability in day of emergence of subsequent follicular waves was lower in cows receiving EB + P4 than in SO-injected cows (P < 0.05). The percentage of cows that ovulated was not different (P = 0.16), but CR increased the percentage of cows that ovulated when groups that received EB + P4 were compared to the EB + P4 group that did not have CR (53.1 vs 11.1%, respectively; P < 0.05). Maximum diameter of dominant follicles was larger (P = 0.05) in ovulatory follicles. The luteal phase was longer (P < 0.03) in cows receiving EB + P4 injection (10.6 +/- 1.2 d) than in cows receiving SO (4.4 +/- 2.2 d). In summary, the maximum size of ovulatory follicles was greater than that of nonovulatory follicles, the ovulatory response of postpartum anestrous cows was maintained through d 8 of a follicular wave, synchronization of follicular waves was accomplished in postpartum cows using EB + P4, and the subsequent luteal phase length was increased in animals that were administered EB + P4.     

Effects of intraovarian infusion of insulin-like growth factor-I on ovarian follicular function in cattle

The objective of this study was to determine if increased secretion of intraovarian insulin-like growth factor-I (IGF-I), experimentally induced via minipumps, affects follicular function in cattle. Fourteen cycling Holstein cows were divided equally into two groups: Control, osmotic minipumps (containing vehicle) surgically inserted into each ovary, or IGF-I treated, osmotic minipumps as in Controls but pumping 2.0 microg of recombinant human IGF-I per hr for 7 days. All cows were synchronized with prostaglandin F(2alpha) 0.10) between Control and IGF-I-treated cows during Days 2 to 6 of treatment. IGF-I treatment increased (P<0.05) estradiol concentrations in follicular fluid of small follicles, but had no effect (P<0.10) on estradiol concentrations in follicular fluid of large follicles, or on progesterone, androstenedione, or IGF binding protein concentrations in small or large follicles. We conclude that a 7-day infusion of IGF-I directly into the stroma of the ovary altered follicular growth and follicular fluid estradiol concentrations.     

Characterization of ovine follicles destined to form subfunctional corpora lutea

A study was done to test whether ovulatory follicles destined to form subfunctional corpora lutea differed from normal ovulatory follicles in steroidogenic function. Twenty-five ewes were treated with prostaglandin F2 alpha on d 11 of the estrous cycle, then unilaterally ovariectomized before (n = 13) or after (n = 12) the surge of luteinizing hormone (LH) at the induced estrus to collect "control" follicles, which would have produced normal corpora lutea. In 15 ewes, the second ovary was removed 63 to 84 h later to collect "treated" follicles before (n = 7) or after (n = 8) the second expected surge of LH. Five ewes (control) were allowed to ovulate from the remaining ovary at first estrus and another five (treated) at the second estrus (3 to 4 d later). Treated ewes had lower serum progesterone than control ewes during the ensuing cycle (P less than .05). Treated follicles contained less estradiol in the theca (4.4 +/- .6 vs 10.0 +/- 2.5 ng; P less than .05), less androstenedione (.1 +/- .1 vs 1.0 +/- .2 ng) and estradiol (.5 +/- .1 vs 2.9 +/- 2.2 ng) in the granulosa (P less than .05) and less progesterone in the follicular fluid (.8 +/- .4 vs 3.3 +/- .8 ng; P less than .05) than control follicles, when removed before the surge of LH. Follicles removed after the surge of LH did not differ. In conclusion, ovulatory follicles with low steroidogenic function became corpora lutea that secreted lower-than-normal quantities of progesterone.