Surges of FSH during the follicular and early luteal phases of the estrous cycle in heifers

Surges of FSH were characterized in each of 12 Holstein heifers using a computerized cycle detector program, and as mean changes averaged over all heifers. Blood samples were collected 6 times a day at 4-h intervals beginning at late diestrus. Concentrations of FSH were adjusted relative to the preovulatory LH peak (Hour 0) and profiled beginning 48 h before and ending 120 h after the LH peak. Peak concentrations of FSH and LH occurred synchronously in 11 of 12 (92%) heifers, and only a 4-h interval separated peak concentrations in the remaining heifer. The FSH surge that was synchronous with the LH surge was designated FSH Surge 1 and was used as a reference to designate other FSH surges. Surge -1 of FSH was detected in 58% of the heifers at mean Hour -21.2, and Surges 2, 3 and 4 were detected in 92%, 92% and 75% of the heifers, respectively, at mean Hours 25.1, 57.8 and 78.7. Mean peak levels and duration of FSH Surges-1, 2, 3 and 4 were significantly lower than for FSH Surge 1. Mean concentrations of FSH significantly increased and decreased before and after the LH peak, resulting from the synchrony between FSH Surge 1 and the LH surge in individual heifers. Additionally, there was a tendency (P < 0.08) for a second and third increase in mean FSH concentrations at Hours 24 and 60, which was attributed to FSH Surges 2 and 3 that occurred in individuals. Peak FSH concentrations of Surge 2 occurred (mean, Hour 25.1) within 8 h of maximal mean concentrations at Hour 24 in 91% of the heifers. Correspondingly, peak FSH concentrations of Surge 3 occurred (mean, Hour 57.8) within 8 h of maximal mean concentrations at Hour 60 in 64% of the heifers. Surges -1 and 4 of FSH occurred less frequently and at various times within and among heifers compared with Surges 1 to 3; therefore, they were not detected as mean increases in FSH concentrations but were masked as a result of concentrations being averaged over all heifers. In summary, FSH surges were detected in individual heifers before and after the combined FSH/LH surge. The interpeak intervals for FSH Surges 1 to 2 (25 h), 2 to 3 (33 h) and 3 to 4 (21 h) suggests a rhythmic nature to the surges.     

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

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

Photoaffinity labeling of pituitary gonadotropin releasing hormone receptors during the rat estrous cycle

A bioactive photoaffinity derivative of gonadotropin releasing hormone was used to identify pituitary gonadotropin releasing hormone receptors at various stages of the rat estrous cycle. Sodium dodecyl sulfate polyacrylamide gel electrophoresis resulted in the identification of a single specific component with an apparent molecular weight of 60,000 daltons throughout the estrous cycle. The amount of radioactivity incorporated into the 60K dalton band in diestrus and proestrus female rats increased 2.5-fold to that of metestrus and estrus female rats. These findings provide additional evidence for the identification of pituitary gonadotropin releasing hormone receptors.     

Periodic fluctuations in FSH concentrations during the ovine estrous cycle

Heterologous radioimmunoassays for ovine LH and ovine FSH were validated and used to examine the concentrations of gonadotropins during the estrous cycle. Concentrations of LH were maximal on the day of estrus as previously reported. Concentrations of FSH were minimal 1 or 2 days before estrus, increased markedly during estrus, and fluctuated widely during diestrus. Most ewes ($\text{11}\text{13}$) had periodic waves of FSH occurring at short intervals (3.5–6 days, $\text{3}\text{13}\text{ewes}$), long intervals (10–18 days, $\text{3}\text{13}\text{ewes}$), or at both long and short intervals ($\text{5}\text{13}\text{ewes}$).     

Dynamic change in charge heterogeneity of pituitary FSH throughout the estrous cycle in female rats

Anterior pituitaries were removed from female rats at various stages during the estrous cycle and FSH was fractionated by isoelectric focusing (IEF). Fourteen FSH components were observed during the estrous cycle and twelve of them were distributed between pH 3.71 and 6.66. IEF profiles of FSH in the pituitaries varied with the stage in the estrous cycle. Especially at the time of serum FSH surge on the day of proestrus, most of the components decreased, while only a highly alkaline component showed an increase. When these FSH components were separated and their nature was examined by radioreceptor assay (RRA), radioimmunoassay (RIA) and gel filtration, differences were observed among these components in the RRA/RIA ratio and gel filtration profile. As a general tendency, the RRA/RIA ratio of the components became greater while the apparent molecular size became smaller, as their pI became higher. However, some highly acidic components showed a biphasic elution pattern and the most acidic one eluted the latest on gel filtration, suggesting that these components may be heterogeneous in terms of molecular size. The FSH concentration in sera collected at different stages in the estrous cycle was measured by both RRA and RIA. The RRA/RIA ratio was high when the serum immunoreactive FSH was low, and low during the FSH surge. From these findings, it is concluded that the quality of FSH molecules present in the anterior pituitary gland changes dynamically throughout the estrous cycle, especially during the period of serum FSH surge. Furthermore it is suggested that the type of FSH secreted from it also varies according to the stage in the estrous cycle.     

Plasma inhibin A in heifers: relationship with follicle dynamics, gonadotropins, and steroids during the estrous cycle and after treatment with bovine follicular fluid

The relationship between follicle growth and plasma inhibin A, FSH, LH, estradiol (E), and progesterone was investigated during the normal bovine estrous cycle and after treatment with steroid-free bovine follicular fluid (bFF) to arrest follicle development. In the first study, four heifers were monitored over three prostaglandin (PG)-synchronized cycles. Blood was collected every 2-8 h, and ovaries were examined daily by ultrasonography. Inhibin A was measured using a modified enzyme-linked immunosorbent assay that employed a new monoclonal antibody against the alpha subunit of bovine inhibin. Plasma inhibin A ( approximately 50 pg/ml before luteolysis) rose steadily during the induced follicular phase (P < 0.05) to a peak ( approximately 125 pg/ml) coincident with the preovulatory E/LH/FSH surge. After ovulation, inhibin A fell sharply (P < 0.05) to a nadir ( approximately 55 pg/ml) coincident with the secondary FSH rise. During the next 3 days, inhibin A increased to approximately 90 pg/ml in association with growth of the new dominant follicle (DF). Plasma E also rose twofold during this period, whereas FSH fell by approximately 50%. Inhibin A was negatively correlated with FSH (r = -0.37, P < 0.001) and positively correlated with E (r = 0.49, P < 0.0001). Observations on eight cycles (two cycles/heifer), in which growth of the ovulatory DF was monitored from emergence to ovulation, showed that the first-wave DF (DF1) ovulated in three cycles and the second-wave DF (DF2) in five cycles. After PG, plasma inhibin A and E increased similarly in both groups, with concomitant falls in FSH. In the former group, the restricted ability of DF1 to secrete both inhibin A and E was restored after luteolysis. Results indicate that dynamic changes in the secretion of both E and inhibin A from the DF contribute to the fall in FSH during the follicular phase and to the generation and termination of the secondary FSH surge, both of which play a key role in follicle selection. In the second study, bFF (two dose levels) was administered to heifers (n = 3-4) for 60 h starting from the time of DF1 emergence. Both doses suppressed FSH (P < 0.05) and blocked DF1 growth to the same extent (P < 0.01), although inhibin A levels were only marginally raised by the lower dose (not significant compared to controls). The high bFF dose raised (P < 0.001) inhibin A to supraphysiological levels ( approximately 1 ng/ml). A large "rebound" rise in FSH occurred within 1 day of stopping both treatments, even though the inhibin A level in the high-dose bFF group was still approximately threefold higher than that in controls. This indicates that desensitization of gonadotropes to inhibin negative feedback is a contributory factor, together with reduced ovarian output of E, in generation of the post-bFF rebound in FSH.     

Analysis of urinary immunoreactive steroid metabolites and gonadotropins for characterization of the estrous cycle,breeding period,and seasonal estrous activity of captive killer whales (Orcinus orca)

To increase the basic understanding of killer whale (Orcinus orca) reproductive physiology necessary for the development of artificial breeding programs, we utilized radioimmunoassays (RIA) to detect urinary immunoreactive steroid metabolites (pregnanediol-3α-glucuronide [PdG] and estrone-conjugates [EC]) and gonadotropins (luteinizing hormone [LH] and follicle-stimulating hormone [FSH]) in urine samples from six female killer whales. Urine samples were collected from the whales by voluntary presentation behavior over a 2- to 4-year period. All urinary hormone values were corrected for intersample urine concentration variations by indexing with creatinine. Daily urine samples from four whales were collected during two conceptions and 18 complete estrous cycles. LH, FSH, EC, and PdG immunoreactive levels were determined and combined with observed copulatory activity in five cycles, including two conceptive cycles from two whales. Mean luteal phase lengths ranged from 9.7 to 19.2 days. Mean follicular phase lengths ranged from 6.5 to 16.8 days. Mean estrous cycle lengths based on the first detectable PdG levels were 41.6 ± 6.72 S.E.M. days. After PdG nadir, immunoreactive FSH levels showed a bimodal pattern with the first peak being greater in size, and both preceding a follicular phase EC increase. LH levels > the 95% confidence interval of the mean were considered significant. Combined LH immunoreactive values from whales 2 and 6 during two and three estrous cycles, respectively, had significant LH peak concentrations on day minus 2. These significant LH peaks were assumed to represent the preovulatory LH surge. Eight copulations during two conceptive cycles were observed between whales 2 and 6 and a breeding male. Six of these copulations (3 with each female whale) occurred within 72 hours of the beginning or the end of the presumptive preovulatory LH surge. Estrous activity was seen throughout the year for the herd. However, individuals had varying periods of anestrus that could not be linked to environmental, social, or nutritional influences. The whales that were reproductively successful had anestrus intervals that were usually influenced by gestation, postparturient period, or lactation. The information obtained during this research enhances the foundation for future artificial reproductive management techniques. © 1993 Wiley-Liss, Inc.     

Uterine steroid hormone receptors during the estrous cycle and during hibernation in the Turkish hamster (Mesocricetus brandti)

The Turkish hamster is a long-day breeder that hibernates for 4-5 mo if exposed to a short-day, cold environment. The objective of this study was to assess the uterine responsiveness of the hibernating animal to ovarian steroids. Our approach was 1) to characterize and determine uterine estrogen (E) and progesterone (P) receptors (R) during hibernation as compared to the levels observed in cycling females that had terminated hibernation, and 2) to assess the responsiveness of the uterus to E during hibernation by its ability to induce uterine P receptor. Females were exposed to short days (10L:14D) for 2 mo and then were placed in a cold-room (10L: 14D, 6 +/- 1 degrees C). After 2 or 4 mo in the cold, hibernating animals were killed and uterine steroid receptors were determined by 3H-steroid binding assay. Uterine receptors were also determined in cycling Turkish hamsters on each morning of the estrous cycle. Values for uterine receptors (pmol/g tissue, n = 4-6) during the estrous cycle (estrus, diestrus I, diestrus II, proestrus) were: 4.3 +/- 0.78, 3.9 +/- 0.19, 4.1 +/- 0.25, 3.7 +/- 0.5 for cytosolic ER; 36.6 +/- 5.8, 32.2 +/- 6.8, 36.3 +/- 1.5, 54.4 +/- 1.9 for cytosolic PR; 0.59 +/- 0.11, 0.54 +/- 0.07, 1.06 +/- 0.05, 1.42 +/- 0.17 for nuclear ER. Hibernating (torpid) animals sampled after 2 mo in the cold showed a significant (p less than 0.05) depression of cytosolic ER (2.6 +/- 0.12, n = 5) and cytosolic PR (19.0 +/- 2.6, n = 8) as compared to any day of the estrous cycle.(ABSTRACT TRUNCATED AT 250 WORDS)     

Associations between emergence of follicular waves and fluctuations in FSH concentrations during the estrous cycle in ewes

Folliculogenesis was studied daily in 16 interovulatory intervals in 5 Polypay ewes from mid February through April using transrectal ultrasonic imaging. The 3-mm follicles attaining > or = 5 mm on Days--1 (ovulation=Day 0) to 11 showed significant peak numbers on Days 0, 5 and 10. The number of 3- and 4-mm follicles that did not reach > 4 mm was not significant, indicating that these follicles did not manifest a wave pattern. A follicular wave was defined as one or more follicles growing to > or = 5 mm; the day the follicles were 3 mm was the day of wave emergence, and the first wave after ovulation was Wave 1. Waves 1, 2 and 3 emerged on Days -1 to 2,4 to 7 and 8 to 10, respectively. Four interovulatory intervals in April were short (9 to 14 d), indicating the end of the ovulatory season. In the remaining 12 intervals, the ovulatory wave was Wave 3 in one interval, Wave 4 in 8 intervals, and Wave 5 or 6 in 3 intervals. The ovulatory wave followed the rhythmic pattern of Waves 1 to 3 by emerging on Days 11 to 14 in 50% of the intervals. In the remaining intervals, either the ovulatory wave was Wave 4 but did not emerge until Day 16 or other waves intervened between Wave 3 and the ovulatory wave. The longest intervals (22, 24 and 24 d) had >4 waves. Based on a cycle-detection program, peak values of FSH fluctuations were temporally associated with the emergence of waves as indicated by the following: 1) tendency (P < 0.08) for an increase in FSH concentrations between 3 and 2 days before emergence of a wave; 2) close agreement between mean number of waves per interval (mean +/- SEM, 4.1 +/- 0.3) and mean number of identified FSH fluctuations (4.5 +/- 0.3); 3) close agreement in length of interwave intervals (4.0 +/- 0.3) and interpeak (FSH) intervals (3.6 +/- 0.2); 4) positive correlation (r(2)=0.8) for number of the 2 events (follicular waves and FSH fluctuations) within intervals; and 5) a closer (P < 0.01) temporal relationship between the 2 events than would have been expected if the events were independent. The results support a relationship between transient increases in FSH concentrations and emergence of follicular waves throughout the interovulatory interval in Polypay ewes, with the 2 events occurring approximately every 4 d.     

Superovulation in heifers given FSH initiated either at day 2 or day 10 of the estrous cycle

The aim of this study was to determine if initiation of superovulation in heifers during the time of development of the first dominant follicle (Days 2 to 6) would give equivalent ovulation and embryo production rates as treatment initiated at mid-cycle. Estrus was synchronized in 60 beef heifers using luprostiol (PG) and they were randomly allocated to treatment with 4.5, 3.5, 2.5 and 1.5 mg of porcine follicle stimulating hormone (FSH) administered twice daily, either on Days 2, 4, 5 and 6 (Day-2 group), respectively, or with similar doses at four consecutive days during mid-cycle (Day-10 group, initiation on Day 9 to 11). All heifers received 500 mug cloprostenol at the fifth FSH injection and 250 mug at the sixth injection. Blood samples for progesterone determination were collected at the time of FSH injections. Heifers were slaughtered 7 d post estrus, and the number of ovulations and large follicles (>/=10mm) were determined on visual inspection of the ovary. Following flushing of the uterine horns the quality of embryos and the fertilization rate were determined. Significant differences between treatments were determined using a two-sided t-test, and frequency distributions were compared using Chi-square tests. The mean number (+/-SEM) of ovulations for heifers in the Day-10 group was 12.9+/-1.0, and 8.5+/-0.9 embryos were recovered. Both the number of ovulations (6.7+/-0.8) and embryos recovered (4.1+/-0.6) were lower (P=0.0001) in heifers in the Day-2 group. Following grading based on a morphological basis, a higher number (P=0.002) of embryos was categorized as Grades 1 and 2 (4.1+/-0.6) and Grade 3 (2.1+/-0.4) in Day-10 heifers than in the Day-2 group (Grade 1 and 2, 1.9+/-0.3; Grade 3, 0.7+/-0.2). The number of Grade 4 and 5 embryos (Day 10, 1.6+/-0.2; Day 2, 1.4+/-0.2) and the number of unfertilized ova (Day 10, 0.7+/-0.4; Day 2, 0.2+/-0.1) did not differ between treatments. Progesterone concentrations were lower (P=0.0001) in Day-2 heifers at FSH treatment prior to prostaglandin, and the decline was more rapid following prostaglandin injection at Day 5 (P=0.02). Results of this study indicate that the number of ovulations and embryos recovered was lower in heifers when FSH treatment was initiated on Day 2 compared with Day 10 of the estrous cycle.     

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

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

Follicular populations during the estrous cycle in heifers. I. Influence of day

Ovarian follicles ⩾ 2 mm were studied in 22 Holstein heifers by daily ultrasound examinations. There were significant differences (P < 0.0001) among days of the estrous cycle for diameter of the largest and second largest follicles and in the numbers of follicles 2–3 mm, 4–6 mm, 7–10 mm, 11–13 mm, > 13 mm, and total number of follicles ⩾2 mm. Patterns of the mean profiles for all follicular endpoints except the number of follicles 4–6 mm and total number of follicles ⩾ 2 mm were bimodal. The days encompassed by the first and second portions, respectively, of the bimodal profiles were approximately: diameter of largest follicle, Days 0–14 and 15–21 (ovulation); diameter of second largest follicle, Days 0–7 and 8–20; number of follicles 2–3 mm, Days 1–11 and 12–20; number of follicles 7–10 mm, Days 0–6 and 7–18; number of follicles 11–13 mm, Days 0–8 and 9–20; and number of follicles > 13 mm, Days 2–14 and 16–21. Data for the various categories were recombined to demonstrate relationships between the numbers of follicles 2–3 mm and ⩾ 4 mm during the interovulatory interval. There were significant differences (P < 0.0001) among days in both 2–3 mm and ⩾ 4 mm follicular categories. Differences appeared due to periods of higher mean numbers of follicles 2–3 mm which began between Days 2 or 3 and Days 15 or 16 and reached maximum levels on Day 7 and Day 19, respectively. There was an inverse relationship between the number of follicles 2–3 mm vs ⩾ 4 mm and between the diameters of largest and second largest follicles. The process of selection of the follicle destined to ovulate appeared to become manifest as selective growth of the preovulatory follicle with concurrent decrease in diameter of the second largest follicle and regression of the other follicles in the various follicular categories. A similar process apparently occurred early in the interovulatory interval. There was apparently selective growth of a follicle to preovulatory size by Day 6, coincident decrease in diameter of the second largest follicle, and apparent regression of other follicles in the ultrasonically detectable pool. The only apparent difference was that the follicle which attained preovulatory diameter early in the interovulatory interval remained in the ovary for 5 or 6 days, then regressed, while the follicle which attained preovulatory diameter at approximately Day 18–20 ovulated.     

Relationship between opposite changes of vaginal and vestibular impedance during estrous cycle in sows

Several bioimpedance techniques have been developed for noninvasive monitoring of reproductive events occurring in cyclic gilts and sows. Our objective was to compare the changes of vaginal and vestibular impedance during the porcine estrous cycle (experiment 1). In addition, we examined the causes of impedance variations in the vaginal vestibule during periestrus (experiment 2). The vaginal and vestibular impedance were measured with specially designed instruments. Sows were monitored for estrus via exposure to a sexually mature boar. The impedance in the vagina decreased gradually after weaning (P<0.01) reaching its nadir 2 days before estrus and increased during estrus to near maximum 2 days after estrus onset (P<0.01). The vaginal impedance during diestrus reached approximately the same level as on the weaning day. In contrast, the impedance in the vaginal vestibule increased gradually after weaning, then markedly during estrus (P<0.01) reaching its maximum 2 days after the onset of estrus followed by an abrupt decrease during early diestrus (P<0.01). The vestibular impedance after early diestrus reached almost the same level as before estrus. A significant negative correlation was found between the vaginal impedance in proestrus and vestibular impedance in periestrus. In experiment 2, interaction of the interval from weaning to estrus and parity significantly influenced the vestibular impedance in periestrus. The breed of sows did not affect the impedance in the vaginal vestibule through the whole experiment. From the present study we conclude that closely related inverse changes of the vaginal and vestibular impedance take place in pigs during the estrous cycle.     

Repeatability of blood serum progesterone levels in dairy heifers on day 7 of the estrous cycle

Thirty normally cycling dairy heifers were used to determine the repeatability of blood serum progesterone levels on Day 7 ± 0.25 d of the estrous cycle. The experimental group consisted of 16 Holsteins and 14 dairy crossbreds ranging in age from 18 to 24 months. Day of the estrous cycle was determined from twice daily observations for standing heat (Day 0). Serum progesterone levels for Day 7 ± 0.25 d were determined by radioimmunoassay from blood samples collected by jugular venipuncture over three to four consecutive estrous cycles. Levels of blood serum progesterone for Day 7 ± 0.25 d ranged from 0.57 to 6.03 ng/ml. Least square means for the Holstein (2.74 ng/ml) and dairy crossbred (3.38 ng/ml) groups were different (P<0.006). The repeatability for levels of blood serum progesterone on Day 7 of the estrous cycle was low (0.0115).