Relationship between days of the luteolytic period and locations of the preovulatory follicle and CL in interovulatory intervals with two or three follicular waves in heifers

Diameter of follicles was determined every 12 hours and progesterone (P4), FSH, and LH concentrations were determined every 6 hours from Day 12 (Day 0 = ovulation) to the ovulation at the end of the interovulatory interval (IOI). Groups were assigned on the basis of an ipsilateral (Ipsi) versus contralateral (Contra) relationship between the preovulatory follicle and CL and two follicular waves (2W) versus three waves (3W) per IOI. Numbers of IOIs were Ipsi-2W (n = 6), Ipsi-3W (n = 6), and Contra-3W (n = 8). Normalization to the end of luteolysis (day that P4 was closest to 1.0 ng/mL) indicated for the first time that concentrations of P4 and FSH were greater (P < 0.05) in 3W IOIs than in 2W IOIs for the 3 days before the beginning of a P4 decrease. The beginning of a P4 decrease occurred about 5 days and 6 hours after emergence of the preovulatory wave at 6 mm in 2W and 3W IOIs, respectively. On the day of diameter deviation between the future dominant and largest subordinate follicles in wave 3 of 3W IOIs, the future dominant follicle had the following characteristics: (1) distribution of diameters differed (P < 0.01) from unimodality; (2) diameter was greater (P < 0.05) in the Contra-3W group (9.8 ± 0.4 mm) than in the Ipsi-3W group (8.8 ± 0.3 mm); (3) diameter was similar to the diameter at the beginning of the P4 decrease (9.6 ± 0.9 mm); and (4) diameter was as small or smaller than diameter of the largest subordinate in seven of 14 heifers compared with zero of seven heifers in wave 2 of 2W IOIs. The differences involving deviation may be related to a reported greater frequency of the Contra-3W group than Ipsi-3W group. Results supported the hypothesis that emergence of the ovulatory wave occurs well before the beginning of luteolysis in 2W IOIs and near the beginning of luteolysis in 3W IOIs.     

Complex interrelationships among CL,preovulatory follicle,number of follicular waves,and right or left ovaries in heifers

The diameter of the dominant follicle (DF) of wave 1 was studied on Days 9 to 17 (Day 0 = ovulation) in a survey of the ipsilateral and contralateral relationships between the location of the DF and CL, and number of follicular waves per interovulatory interval (IOI). For contralateral relationships, regardless of number of waves the diameter of the DF of wave 1 decreased (P < 0.03) between Days 11 and 13 when referenced to the follicle–CL relationship of wave 1 and decreased (P < 0.008) between Days 9 and 11 when referenced to the preovulatory follicle (PF)–CL relationship. For wave 2 in two-wave IOIs, the CL ovary of ipsilateral relationships had more (P < 0.05) follicles that reached at least 6 mm than the non-CL ovary. In three-wave IOIs, frequency of IOIs with the DF in the CL ovary was greater (P < 0.02) for wave 2 than for wave 3. In wave 3, the preovulatory and the largest subordinate follicles were located more frequently (P < 0.005) in the contralateral ovary. Ovulation in two-wave IOIs occurred more frequently (P < 0.0009) from the right ovary. In three-wave IOIs with a contralateral relationship ovulation occurred more frequently (P < 0.003) from the left ovary; a negative intraovarian effect of the CL on location of the PF may account for more ovulations from the left ovary and a reported greater frequency of the contralateral relationship. The hypothesis was supported that the ipsilateral versus contralateral relationship between the PF and CL is affected by the DF–CL relationship during the previous follicular waves and by the number and identity of waves per IOI.     

Minor FSH surge,minor follicular wave,and resurgence of preovulatory follicle several days before ovulation in heifers

Blood samples were collected and follicle diameters were determined daily beginning on Day 12 (Day 0 = ovulation) in 35 interovulatory intervals (IOIs) in heifers. A minor follicular wave with maximal diameter (6.0 ± 0.3 mm) on Day −4 was detected in six of seven IOIs that were scanned for follicles 4 mm or greater. The number of IOIs with a CV-identified minor FSH surge toward the end of the IOI was greater (P < 0.03) in two-wave IOIs (10/17) than in three-wave IOIs (4/18). The 17 two-wave IOIs were used for study of the temporal relationships among preovulatory follicle, FSH, LH, and estradiol. Daily growth rate of the preovulatory follicle was maximum on Days −11 to −7, minimum (P < 0.05) on Days −7 to −4, and increased (resurged, P < 0.05) on Days −4 to −3. A transient increase in FSH was maximum on mean Day −4, and the peak of a minor FSH surge occurred on Day −4.5 ± 0.2. Concentration of LH and estradiol increased between Days −5 and −4. Results demonstrated resurgence of the preovulatory follicle apparently for the first time in any species. Resurgence seemed more related temporally to the minor FSH surge than to the LH increase, but further study is needed. Results supported the novel hypotheses that a minor FSH surge near the end of the IOI is temporally associated with (1) the emergence of a minor follicular wave and (2) the resurgence in growth rate of the preovulatory follicle.     

Hormone concentrations temporally associated with contralateral and ipsilateral relationships between the CL and preovulatory follicle during the third follicular wave in heifers

Concentrations of circulating hormones after Day 14 (Day 0 = ovulation) were determined daily in 87 interovulatory intervals (IOIs) in heifers. The IOIs were grouped into four permutations according to an ipsilateral (Ipsi) or contralateral (Contra) relationship between the CL and the preovulatory follicle and two (2W) or three (3W) follicular waves per IOI. The number of IOIs per group differed (P < 0.005) from equality among the Ipsi-2W (n = 27), Contra-2W (n = 31), Ipsi-3W (n = 9), and Contra-3W (n = 20) groups. A continuous decrease in progesterone (luteolysis) began later (P < 0.05) in the Contra-3W group (Day 18.0 ± 0.4) than in each of the Ipsi-2W (15.4 ± 0.2), Contra-2W (15.6 ± 0.2), and Ipsi-3W (16.2 ± 0.5) groups. Concentrations of LH and estradiol began to increase near the beginning of luteolysis in each group. A minor FSH surge that did not stimulate a major follicular wave developed in about 50% of the IOIs in each group, except that none were detected in the Ipsi-3W group. The minor FSH surge reached a peak about 4 days before ovulation and several days after wave 3 had emerged. The hypothesis that luteolysis begins earliest in two-wave IOIs, intermediate in three-wave IOIs with an ipsilateral CL/follicle relationship, and latest in three-wave IOIs with a contralateral relationship was supported. The hypothesis that a minor FSH surge occurs most frequently in association with three follicular waves was not supported.     

Evaluation of the ultrasound image attributes of developing ovarian follicles in the four follicular waves of the interovulatory interval in ewes

Computer-assisted quantitative echotextural analysis was applied to ultrasound images of antral follicles in the follicular waves of an interovulatory interval in sheep. The ewe has three or four waves per cycle. Seven healthy, cyclic Western White Face ewes (Ovis aris) underwent daily, transrectal, ovarian ultrasonography for an interovulatory interval. Follicles in the third wave of the ovulatory interval had a longer static phase than that of those in Waves 1 and 2 (P < 0.05). The numeric pixel value for the wall of anovulatory follicles emerging in the third wave of the cycle was significantly higher than that for Waves 1 and 2 at the time of emergence (156.7 ± 8.09, 101.6 ± 3.72, and 116.5 ± 13.93, respectively), and it decreased as follicles in Wave 3 reached maximum follicular diameter (P < 0.05). The numeric pixel value of the antrum in the ovulatory follicles decreased as follicular diameter increased to ≥5 mm in diameter (P < 0.05). The pixel heterogeneity of the follicular antrum in Wave 1 increased from the end of the growth phase to the end of the regression phase for follicles in that wave (P < 0.05). The total area for the wall and antrum of the follicles studied were correlated with follicular diameter in all follicular waves (r = 0.938, P < 0.01 and r = 0.941, P < 0.01 for the wall and antrum, respectively). Changes in image attributes of the follicular wall and antrum indicate potential morphologic and functional differences among antral follicles emerging at different stages of the interovulatory interval in cyclic ewes.     

Characteristics of peaks in serum concentrations of follicle-stimulating hormone and estradiol, and follicular wave dynamics during the interovulatory interval in cyclic ewes

There are three or four ovarian follicular waves in the interovulatory interval of cyclic ewes. Each follicular wave is preceded by a transient peak in serum follicle-stimulating hormone (FSH) concentrations. Serum concentrations of estradiol also increase concurrent with the growth of follicle(s) in each wave. In the current study, we investigated the patterns of follicular wave development and characteristics of FSH and estradiol peaks in all follicular waves of the interovulatory interval and after induction of a supraphysiologic FSH peak in cyclic ewes (Ovis aris). In Experiment 1, 19 ewes underwent daily ovarian ultrasonography and blood sampling for a complete interovulatory interval. In Experiment 2, seven ewes received two administrations of ovine FSH (oFSH), 8 h apart (1 μg/kg; sc), at the expected time of the endogenous FSH peak preceding the second follicular wave of the interovulatory interval. In Experiment 1, the amplitude of the FSH peaks decreased (up to 50%), whereas basal serum FSH concentrations increased across the interovulatory interval (P < 0.05). Maximum follicular diameter was greater (P < 0.05) for Wave 1 and the Ovulatory wave (6.0 ± 0.3 and 6.1 ± 0.2 mm, respectively) than for Waves 2 and 3 (5.3 ± 0.1 and 5.4 ± 0.3 mm, respectively). Life span was greater for follicles in Wave 1 compared with other waves (P < 0.05). Treatment with oFSH increased the amplitude of an FSH peak by 5- to 6-fold. This treatment increased estradiol production (P < 0.05) but had little effect on other characteristics of the subsequent follicular wave. We concluded that changes in the amplitude and duration of the peaks in serum concentrations of FSH that precede follicular waves across the interovulatory interval do not influence the characteristics of the follicular waves that follow.     

Influences of diet during gestation on potential postpartum reproductive performance and milk production of beef heifers

The influences of nutritional protein and energy during early and mid pregnancy on milk production and postpartum reproductive parameters were determined in 70 beef heifers of two composite breeds (Bos indicus X Bos taurus). At artificial insemination (AI), heifers were divided into four dietary treatment groups identified by the level of protein, and to a lesser extent energy, fed during the first and second trimesters: high/high (HH), high/low (HL), low/high (LH), and low/low (LL). Milk production was lower in the heifers receiving high treatment in first trimester than that in heifers receiving the low treatment (P = 0.01). Milk production was negatively associated with dam body condition score (BCS; P = 0.01), nonesterified fatty acids (P = 0.001), and leptin (P = 0.02) and positively associated with urea (P < 0.001) concentrations during lactation. Increased dietary protein in the first trimester increased or decreased concentrations of colostral protein dependent upon genotype (P = 0.03). Colostral protein was positively associated with bovine pregnancy associated glycoprotein from late gestation (P = 0.007). Milk fat was negatively associated with BCS (P = 0.007) and influenced by genotype (P = 0.003). Dietary treatment did not affect the postpartum reproductive performance of beef heifers. Gestation length (P < 0.001) and the postpartum interval to first estrus (PPI; P = 0.02) were positively associated with calf size. Placental size was negatively associated with placental expulsion time (P < 0.01). Prepartum BCS of the heifers was negatively associated with PPI (P = 0.01). Overall, high levels of nutrition during early gestation are detrimental to milk production in beef heifers.     

Embryo loss in cattle between Days 7 and 16 of pregnancy

Embryo loss between embryonic Days 7 and 16 (Day 0 = day of IVF) in nonlactating cattle, Bos taurus, was analyzed using transfer of 2449 (in groups of 3 to 30) in vitro-produced (IVP) blastocysts. In 152 transfers, pregnancy losses attributable solely to recipient failings amounted to between 6% (beef heifers) and 16% (parous dairy cows), of which 3% were caused by uterine infections. Neither season, year, nor the age of the embryos on retrieval affected pregnancy rates. The latter observation indicated that the reason that a recipient failed to retain embryos was already present at the time of transfer. Notably, the proportion of embryos recovered decreased (P = 0.03) as more embryos were transferred, particularly at later stages (Day 14, P < 0.01). The average length of embryos decreased by approximately 5% for every additional embryo transferred (P < 0.0001). These effects may be linked to embryonic migration. Embryo mortality inherent to the embryo during the second week of pregnancy was 24%. Additionally, 9% of Day 14 embryos were of inferior quality, as they did not contain an epiblast. Combining embryo and recipient causes but excluding infection effects, embryonic loss of IVP embryos during the second week of pregnancy amounted to 26% (heifers) or 34% (parous dairy cows). The length of embryos doubled every day between Days 9 and 16, with a 4.4-fold range in sizes representing two thirds of the variation in length. Embryos retrieved from heifers were twice the size of those incubated in parous cows (P < 0.0001), indicating faster embryonic development/trophoblast proliferation in heifers. Whereas season did not affect embryo recoveries, length was lower (50%) in winter (winter-autumn, P < 0.05; winter-spring, P < 0.001). Lastly, transuterine migration in cattle, when transferring multiple embryos, commenced at Day 14 (4%) and had occurred in all recipients by Day 16 (38% of embryos found contralaterally).     

Suppression of dominant and subordinate ovarian follicles by a proteinaceous fraction of follicular fluid in heifers

Nulliparous Holstein heifers were examined ultrasonically once daily during an interovulatory interval (ovulation = Day 0). Follicles with a diameter >/=4 mm were sequentially identified. Heifers were randomized into four groups (n = 4 heifers per group): untreated control heifers and those treated on Days 0 to 3, Days 3 to 6, or Days 6 to 11. Heifers designated for treatment were given an intravenous injection, twice daily, of a proteinaceous fraction of follicular fluid (PFFF; 16 ml) prepared by extracting bovine follicular fluid with activated charcoal. Mean cessation of growth of the dominant follicle of Wave 1 was later (P<0.005) in control heifers (Day 5.5) than in heifers treated on Days 0 to 3 (Day 1.5) or Days 3 to 6 (Day 3.5). Mean onset of regression of the dominant follicle of Wave 1 was later (P<0.005) in control heifers (Day 12.0) than in heifers treated on Days 0 to 3 (Day 5.0) or Days 3 to 6 (Day 7.5). Mean cessation of growth of the largest subordinate follicle of Wave 1 was later (P<0.05) in control heifers (Day 3.0) than in heifers treated on Days 0 to 3 (Day 1.2). Mean onset of regression of the largest subordinate follicle of Wave 1 was later (P<0.05) in control heifers (Day 7.0) than in heifers treated on Days 0 to 3 (Day 4.8). In heifers treated on Days 6 to 11, cessation of growth and onset of regression of the dominant follicle (means, Days 5.2 and 12.0, respectively) were not significantly different from those of the controls. The hypothesis that PFFF treatment on Days 0 to 3 would cause suppression of all follicles of Wave 1 was supported. The hypothesis that PFFF treatment on Days 3 to 6 would not alter growth of the dominant follicle of Wave 1 was not supported. The mean day of detection of the dominant follicle of Wave 2 was different (P<0.005) in control heifers (Day 8.5) than in heifers treated on Day 0 to 3 (Day 5.5) or Days 6 to 11 (Day 14.2). The mean length of the interovulatory interval was shorter (P<0.05) in control heifers (20.5 d) than in heifers treated on Days 6 to 11 (23.2 d). The hypothesis that PFFF treatment on Days 6 to 11 would delay the emergence of Wave 2 was supported. The proportion of heifers with 2-wave interovulatory intervals was 3 4 for control heifers and 0 4 , 1 4 , and 4 4 for heifers treated on Days 0 to 3, Days 3 to 6, and Days 6 to 11, respectively (3 4 vs 0 4 , P<0.05); the remaining heifers had 3-wave interovulatory intervals. On average, in PFFF-treated heifers, follicles stopped growing 1 d after treatment was started, and Wave 2 was detected 3 d after treatment was stopped.     

Effect of growth and development during the rearing period on the subsequent fertility of nulliparous Holstein-Friesian heifers

This study investigated the effect of growth parameters and metabolic indices during the rearing period on the fertility of nulliparous Holstein-Friesian heifers managed on 17 UK dairy farms. Growth parameters (body weight [BW], heart girth, height, and crown-rump length) and metabolic indices (insulin-like growth factor-I [IGF-I], insulin, glucose, and urea) were measured at approximately 30, 180, and 450 d of age. Fertility data collected included age at first breeding (AFB), number of services per conception, pregnancy rate to first artificial insemination (AI), and age at first calving (AFC). Of the heifers initially bred (n = 428), 4% failed to conceive. The mean pregnancy rate to first AI for heifers that conceived and calved without suffering reproductive loss (n = 392) was 67%, and 6% required >2 inseminations. The mean AFB and AFC was 473 ± 5 d (range, 357 to 936 d) and 791 ± 6 d (range, 636 to 1529 d), respectively. Increased BW, girth, and IGF-I concentration (at 30, 180, and 450 d) and increased skeletal growth (at 180 and 450 d) was associated with a reduced AFB and AFC (P < 0.05 to 0.001). Heifers calving at <775 d had a mean BW gain of 0.82 ± 0.01 kg from 30 to 180 d. Increased glucose concentration at 180 d was associated with a reduced AFB (P < 0.01), but no associations were found between insulin and urea concentrations and any of the fertility traits recorded (P > 0.1). Suboptimal growth associated with an increased AFC could be alleviated by improved monitoring of replacement heifers during the rearing period.     

Changes in the gene expression of adiponectin and adiponectin receptors (AdipoR1 and AdipoR2) in ovarian follicular cells of dairy cow at different stages of development

Adiponectin is one of the most important, recently discovered adipocytokines that acts at various levels to control male and female fertility through central effects on the hypothalamus-pituitary axis or through peripheral effects on the ovary, uterus, and embryo. We studied simultaneous changes in the gene expression pattern of adiponectin and adiponectin receptors 1 and 2 (AdipoR1 and AdipoR2) in granulosa and theca cells, cumulus-oocyte complex, and in corpus luteum in healthy bovine (Bos tarus) follicles at different stages of development. The expression levels of adiponectin, AdipoR1, and AdipoR2 mRNA were lower (P < 0.05) in granulosa and cumulus cells in comparison with that in theca cells and oocyte. In contrast with the oocyte, AdipoR1 in granulosa, theca, and luteal cells was expressed (P < 0.05) more than AdipoR2. Adiponectin expression increased (P < 0.05) in granulosa cells and in cumulus-oocyte complex during follicular development from small to large follicles. Opposite results were observed in theca cells. Expression of adiponectin was highest in the late stages of corpus luteum (CL) regression, whereas lower expression was recorded in active CL (P < 0.05). AdipoR1 and AdipoR2 expression increased during the terminal follicular growth in granulosa and theca cells (P < 0.05) and during the luteal phase progress in CL. There was positive correlation between adiponectin mRNA level in granulosa cells from large follicles and follicular fluid estradiol concentration (r = 0.48, P < 0.05) and negative correlation between adiponectin mRNA abundance in theca cells and follicular fluid progesterone concentration (r = -0.44, P < 0.05). In conclusion, we found that the physiologic status of the ovary has significant effects on the natural expression patterns of adiponectin and its receptors in follicular and luteal cells of bovine ovary.     

Effect of purified llama ovulation-inducing factor (OIF) on ovarian function in cattle

Two experiments were designed to determine the effect of purified ovulation inducing factor (OIF) on ovarian function in cattle. In Experiment 1, prepubertal heifers (n = 11 per group) were treated on Day 5 (Day 0 = day of follicular wave emergence) of the follicular wave with an intramuscular dose of saline (1 mL), GnRH (100 μg), or purified OIF (1 mg/100 kg body weight). Ovulation occurred in 9/11 heifers treated with GnRH, and 1/11 heifers in each of the OIF- and saline-treated groups (P < 0.05). Compared to saline-treated controls, OIF treatment was associated with a smaller dominant follicle diameter (P < 0.01), a rise in plasma FSH concentration (P < 0.1), and earlier emergence of the next follicular wave (P < 0.05). In Experiment 2, sexually mature heifers were given either GnRH or purified OIF on Days 3, 6 or 9 of the first follicular wave (i.e., early growing, early static, or late static phase of the dominant follicle; n = 5 per group per day), or were untreated (n = 10). In heifers treated with OIF on Day 6, the dominant follicle diameter profile tended to be smaller than in controls, and was associated with a rise (P < 0.05) in plasma FSH concentrations. A similar rise in FSH was detected after OIF treatment on Day 9. Compared to untreated controls, treatment with OIF and GnRH was associated with a larger CL diameter (Days 3 and 6 groups; P < 0.05) and a greater concentration of plasma progesterone (Days 6 and 9 groups; P < 0.05). Treatment with purified OIF did not induce ovulation in heifers, but hastened new follicular wave emergence in prepubertal heifers, influenced follicular dynamics in a phase-specific manner in mature heifers, and was luteotrophic.     

Effects of low versus physiologic plasma progesterone concentrations on ovarian follicular development and fertility in beef cattle

The objective of this study was to determine the effects of low versus physiologic plasma progesterone concentrations during the ovulatory wave on fertility in cattle. Suckled beef cows (Bos taurus; n = 129) and pubertal heifers (Bos taurus; n = 150) at random stages of the estrous cycle were given a luteolytic dose of prostaglandin F_2α (500 μg cloprostenol; PGF) twice, 11 d apart. Ten days after the second PGF treatment, cattle were given estradiol benzoate im (1.5 and 1.0 mg for cows and heifers, respectively) and a progesterone-releasing intravaginal device (Cue-Mate) with a single pod containing 0.78 g progesterone (Day 0). Cattle in the low-progesterone group (n = 148) received a luteolytic dose of PGF on Day 0, whereas those in the high-progesterone (i.e., physiologic plasma concentrations) group (n = 131) were allowed to retain their corpora lutea. On Day 8, the Cue-Mate was removed, and PGF was given to both groups. Fifty-four hours to 56 h later, cattle received 12.5 mg of porcine LH (pLH) im and were concurrently artificially inseminated. The dominant follicle in the low-progesterone group was larger (P < 0.001) than that in the high-progesterone group on the day of insemination (14.9 ± 0.3 mm vs. 12.7 ± 0.3 mm, mean ± SEM). At 7 d after ovulation, the low-progesterone group had a larger corpus luteum (24.5 ± 0.54 mm vs. 21.9 ± 0.64 mm, P < 0.01) and higher plasma progesterone concentration (4.0 ± 0.3 vs. 3.1 ± 0.2, P < 0.01) than that of the high-progesterone group. However, pregnancy rates did not differ (79 of 148, 53.4%, and 70 of 131, 53.4%) for low- and high-progesterone groups, respectively). In summary, low circulating progesterone concentrations during the growing phase of the ovulatory follicle resulted in a larger dominant follicle and a larger CL that produced more progesterone, with no significant effect on pregnancy rate.     

Ovarian antral follicular dynamics in sheep revisited: Comparison among estrous cycles with three or four follicular waves

In this study, the characteristics of ovarian follicular waves and patterns of serum concentrations of follicle-stimulating hormone (FSH), estradiol, and progesterone were compared between cycles with three (n = 9) or four (n = 10) follicular waves in Western White Face (WWF) ewes (Ovis aries). Transrectal ultrasonography and blood sampling were performed daily during one cycle. Estrous cycles were 17.11 ± 0.3 and 17.20 ± 0.2 d long in cycles with three and four waves, respectively (P > 0.05). The first interwave interval and the interval from the emergence of the final wave to the day of ovulation were longer in cycles with three waves compared with those in cycles with four waves (P < 0.05). The growth phase (5.1 ± 0.5 vs. 3.1 ± 0.4 d) and life span (5.67 ± 0.3 vs. 4.3 ± 0.3 d) of the largest follicle growing in the last or ovulatory wave was longer in cycles with three waves compared with that in cycles with four waves (P < 0.05). The maximum diameter of the largest follicle was greater in the first wave and the ovulatory wave compared with that in other waves of the cycle (P < 0.05). The regression phase of the largest follicle growing in the first wave was longer in cycles with three waves compared with that in cycles with four waves (4.44 ± 0.4 vs. 3.4 ± 0.4 d; P < 0.05). The length of the life span, regression phase, and, although not significant in every case, FSH peak concentration and amplitude decreased across the cycle (P < 0.05). We concluded that estrous cycles with three or four follicular waves were confined within the same length of cycle in WWF ewes. In this study, there were no apparent endocrine or follicular characteristics that could explain the regulation of the different number of follicular waves (three vs. four) during cycles of similar length.     

Effect of length of progesterone exposure during ovulatory wave development on pregnancy rate

The objective was to determine the effects of the duration of progesterone exposure during the ovulatory wave on fertility (pregnancy rate) in beef cattle. We tested the hypothesis that short-progesterone exposure during the growing and early-static phase of the ovulatory follicle (analogous to the ovulatory wave of 3-wave cycles) is associated with higher fertility than a longer duration of exposure (analogous to the ovulatory wave of 2-wave cycles). Three to 5 days after ovulation, beef heifers (n = 172) and suckled beef cows (n = 193) were given an intravaginal progesterone-releasing device (CIDR) and 2.5 mg estradiol - 17β +50 mg progesterone im to induce a new follicular wave. Cattle were allocated to short- or long-progesterone exposure groups (for 3 and 6 d after wave emergence, respectively) after which prostaglandin F_2α was administered and CIDR were removed. Forty-eight hours later, all cattle were given 12.5 mg pLH and artificially inseminated (AI) with frozen-thawed semen. The diameter of the two largest follicles and the corpus luteum were measured by transrectal ultrasonography at CIDR removal, insemination, and 36 h after insemination. Pregnancy diagnosis was done ultrasonically 38 and 65 d post-AI. There was no difference in pregnancy rates in short- vs long-progesterone exposure in heifers (53 vs 47%, P = 0.44) or cows (63 vs 58%, P = 0.51). However, the diameter of the ovulatory follicle at CIDR removal and AI was smaller in short- than in long-progesterone groups (P < 0.02), and larger in cows than in heifers (P < 0.006). In conclusion, short-progesterone exposure during the growing and early-static phase of the ovulatory follicle (similar to 3-wave cycles) was not associated with higher fertility than a longer progesterone exposure (similar to 2-wave cycles).     

Follicle suppression of circulating follicle-stimulating hormone and luteinizing hormone before versus after emergence of the ovulatory wave in mares

The effect of the ovarian follicles on plasma concentrations of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) before versus after the expected emergence of the ovulatory follicular wave was studied on Days 0 to 18 (Day 0 = ovulation) in four groups of mares (n = 6/group). In addition to a control group, all follicles ≥6 mm in diameter were ablated on Days 0.5, 6.5, or 12.5 in a herd of mares with reported emergence at 6 mm of the future ovulatory follicle on mean Day 10.5. Concentrations of FSH were not different between the Day-0.5 or Day-6.5 ablation groups and the corresponding controls. However, ablation on Day 12.5 resulted in an immediate FSH increase (group-by-day interaction, P < 0.003). For LH, ablation on Day 0.5 resulted in an interaction (P < 0.02), partially from lower (P < 0.05) concentrations on each of Days 15.5 to 18.0 than that in the controls, whereas ablation on Days 6.5 or 12.5 did not result in a significant group effect or interaction. Testosterone concentration, but not progesterone or estradiol concentration, was lower (P < 0.04) on Day 2 in the Day-0.5 ablation group than that in the controls. We inferred that follicles did not contain adequate FSH suppressors on Days 0.5 and 6.5 and that they were present only in the Day-12.5 ablation group or after the expected emergence of the ovulatory wave. The hypothesis of an association between low postovulatory concentrations of an ovarian steroid and low concentrations of LH after Day 15 was supported.     

Developmental competence and mRNA expression of preimplantation in vitro-produced embryos from prepubertal and postpubertal cattle and their relationship with apoptosis after intraovarian administration of IGF-1

Recombinant human Insulin-like growth factor-I (hIGF-1) was administered to one ovary of prepubertal and postpubertal cattle to determine its effects on (1) oocyte developmental competence, (2) the expression pattern of six developmentally important genes (GLUT3, GLUT8, AKT1, BCL-XL, BAD, and BAX), and (3) its relationship with apoptosis (female Holstein-Friesian). Oocytes were retrieved from 7- to 10-mo-old prepubertal dairy calves (preP), 11- to 18-mo-old postpubertal heifers (postP), and cows via ultrasound-guided follicular aspiration. Immature oocytes were matured in vitro then fertilized and cultured up to the blastocyst stage. Apoptosis was determined by terminal deoxynucleotidyl transferase nick-end labeling (TUNEL) in 8-d blastocysts. Similar low blastocyst yields were observed in the IGF-1-treated preP group (11.2 ± 2.4%), the control preP group (10.4 ± 3.0%), and in the IGF-1 postP group (10.9 ± 2.3%). These were lower (P ≤ 0.01) compared with the control postP group (21.2 ± 3.8%) and with cows (23 ± 3.7%). The expression profile of the six genes was partly affected by age and IGF-1 treatment. Apoptosis was correlated with the age of the oocyte donors and was increased in blastocysts derived from prepubertal heifers. Results show that apoptosis is a critical feature of the acquisition of developmental competence of oocytes from prepubertal cattle and that IGF-1 did not beneficially affect oocyte developmental competence.     

The effects of exclusion of progesterone or Day 0 GnRH from a GnRH,prostaglandin, GnRH + progesterone program on synchronization of ovulation in pasture-based dairy heifers

This study evaluated the effect of removing the GnRH injection on Day 0 or the progesterone (P4) device from a GnRH, PGF2α, GnRH (GPG) + P4 program on follicular dynamics and synchronization of ovulation in dairy heifers. Friesian and Friesian × Jersey heifers, in autumn 2009 (n = 35) and spring 2010 (n = 38), were randomly allocated to one of three estrus synchronization programs. The first group (GPG + P4) received 100 μg GnRH on Day 0, a P4-releasing intravaginal device from Days 0 to 7, 500 μg PGF2α on Day 7, and 100 μg GnRH on Day 9, followed by fixed-time artificial insemination 16 to 20 hours later. The program for group 2 (GPG) was the same as group 1 with the exclusion of the P4 device. Group 3 (P + G + P4) was treated the same as group 1, except for the absence of the GnRH treatment on Day 0. Ultrasonography was performed on Days 0, 1, 2, 3, and 7 and then at 12 hourly intervals on Days 9 to 11. Dominant follicle size was affected by both treatment and day, and there was also a significant interaction (P < 0.02) between treatment and day. Mean dominant follicle size was larger in the heifers treated with P + G + P4 on Days 1 to 3 than those treated with GPG + P4 (P < 0.02) and, on Day 2, than those treated with GPG (P = 0.005). However, on Day 7, mean dominant follicle size was larger in heifers treated with GPG than heifers treated with P + G + P4 (P = 0.03). The emergence of a new follicular wave was later in heifers treated with P + G + P4 than heifers, which received a GnRH injection on Day 0 (4.3 ± 0.7 days, compared with combined GPG + P4 and GPG 3.0 ± 0.3 days; P = 0.03). The proportion of heifers that ovulated within the first 48 hours after the Day 9 injection of GnRH was not affected by treatment (GPG, 81%; GPG + P4, 84%; and P + G + P4, 100% [including early ovulation]; P = 0.11). The timing of the ovulation was not different between treatments (P = 0.97).     

Characterization of ovarian follicular wave dynamics in women

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

Association between surges of follicle-stimulating hormone and the emergence of follicular waves in heifers.

The effects of ablation of a dominant follicle and treatment with follicular fluid on circulating concentrations of follicle-stimulating hormone (FSH) were studied and the temporal relationships between surges of FSH and follicular waves were studied in heifers with two or three follicular waves/interovulatory interval. Cauterization of the dominant follicle on Day 3 or Day 5 (ovulation on Day 0) (six control and six treated heifers/day) resulted in a surge (P less than 0.05) in FSH beginning the day after cautery. The FSH surge prior to wave 2 (first post-treatment follicular wave) occurred 4 days (Day 3 cautery) and 2 days (Day 5 cautery) before the surge in control groups, corresponding to a 4-day and a 2-day advance in emergence of wave 2 compared with controls. It was concluded that the dominant follicle on Day 3 and Day 5 was associated with the suppression of circulating FSH concentrations. Heifers (n = 4/group) were untreated or treated intravenously with a proteinaceous fraction of bovine follicular fluid on Days 0-3, 3-6, or 6-11. Concentrations of FSH were suppressed (P less than 0.05) for the duration of treatment, regardless of the days of treatment. Cessation of treatment was followed within 1 day by the start of a surge in FSH. The FSH surge prior to wave 2 occurred 2 days earlier (treatment on Days 0-3), 1 day later (treatment on Days 3-6), and 6 days later (treatment on Days 6-11) than in controls, corresponding to an equivalent advance or delay, respectively, in the emergence of wave 2 compared with controls. The results suggest that the effects of exogenous follicular fluid on follicular development were mediated, in whole or in part, by altering plasma FSH concentrations. Control heifers combined for the two experiments were separated into those with 2-wave (n = 11) or 3-wave (n = 5) interovulatory intervals. Two-wave heifers had two FSH surges and 3-wave heifers had three apparent FSH surges during the interovulatory interval. Results of the cautery and follicular fluid experiments indicated that a surge in FSH necessarily preceded the emergence of a wave. The FSH surges in treated and control heifers began 2-4 days before the detectable (ultrasound) emergence of a follicular wave (follicles of 4 and 5 mm), peaked 1 or 2 days before emergence and began to decrease approximately when the follicles of a wave begin to diverge into a dominant follicle and subordinate follicles (follicles 6-7 mm).