Characterization of thymosin alpha 1 and beta 4 during the bovine estrual period: effects of elevated estradiol and progestin

At present, there is a renewed interest in thymic function and its secretions in relation to endocrine control and reproductive function. In an initial experiment, 60 crossbred heifers (18-20 mo) were detected in estrus and assigned to control or FSH superovulatory groups. On Days 7-14 of the subsequent estrous cycle, FSH was administered for 5 days and prostaglandin F2 alpha (PGF2 alpha) was administered at 48 and 60 h after the initial FSH injection. Control animals received only PGF2 alpha injections between Days 9 and 15 of the cycle. Blood samples were collected from all animals at the time of PGF2 alpha injection and every 12 h thereafter to 72 h post PGF2 alpha injection. In a subsequent experiment, 103 crossbred heifers (16-18 mo) were superovulated with FSH and synchronized to estrus with PGF2 alpha administered 60 h after the initial FSH injection. Twenty-eight of the heifers received Norgestomet implants 12 h prior to the initial PGF2 alpha injection to inhibit the LH surge. Blood samples were collected from animals at 12-h intervals until the PGF2 alpha injection and every 6 h thereafter until 108 h post PGF2 alpha treatment. Although thymosin beta 4 concentrations did change over the estrual period, no differences were noted between control and superovulatory animals in the initial experiment even though estradiol concentrations were increased tenfold from the FSH stimulated ovary. In the second experiment, thymosin beta 4 and alpha 1 increased as the estrual period progressed and decreased (p less than 0.05) subsequent to the LH surge. (ABSTRACT TRUNCATED AT 250 WORDS)     

Prostaglandin F2 alpha-induced release of oxytocin from bovine corpora lutea in vitro

Two experiments were conducted to study the in vitro effects of prostaglandins F2 alpha (PGF2 alpha), E2 (PGE2), and luteinizing hormone (LH) on oxytocin (OT) release from bovine luteal tissue. Luteal concentration of OT at different stages of the estrous cycle was also determined. In Experiment 1, sixteen beef heifers were assigned randomly in equal numbers (N = 4) to be killed on Days 4, 8, 12, and 16 of the estrous cycle (Day 0 = day of estrus). Corpora lutea were collected, an aliquot of each was removed for determination of initial OT concentration, and the remainder was sliced and incubated with vehicle (control) or with PGF2 alpha (10 ng/ml), PGE2 (10 ng/ml), or LH (5 ng/ml). Luteal tissue from heifers on Day 4 was sufficient only for determination of initial OT levels. Luteal OT concentrations (ng/g) increased from 414 +/- 84 on Day 4 to 2019 +/- 330 on Day 8 and then declined to 589 +/- 101 on Day 12 and 81 +/- 5 on Day 16. Prostaglandin F2 alpha induced a significant in vitro release of luteal OT (ng.g-1.2h-1) on Day 8 (2257 +/- 167 vs. control 1702 +/- 126) but not on Days 12 or 16 of the cycle. Prostaglandin E2 and LH did not affect OT release at any stage of the cycle studied. In Experiment 2, six heifers were used to investigate the in vitro dose-response relationship of 10, 20, and 40 ng PGF2 alpha/ml of medium on OT release from Day 8 luteal tissue.(ABSTRACT TRUNCATED AT 250 WORDS)     

Metabolic hormone secretion and FSH-induced superovulatory responses of beef heifers fed dietary fat supplements containing predominantly saturated or polyunsaturated fatty acids

Ovulatory responses following FSH treatment were examined in beef heifers fed dietary fat supplements expected to produce differential effects on serum insulin concentrations and follicular recruitment patterns. Twenty-one heifers (n = 7/group) exhibiting regular estrous cycles were assigned randomly to either a control diet or to 1 of 2 fat-supplemented diets consisting of soybean oil (polyunsaturated fatty acids) or animal tallow (saturated fatty acids). The diets were formulated to be isoenergetic and isonitrogenous, and were fed until ovariectomy between experimental Days 35 and 45. Experimental Day 1 was defined for each heifer as the first day all of the treatment diet was consumed. After 20 d of diet consumption, estrous cycles were synchronized with prostaglandin F(2alpha) (PGF(2alpha)), and ovarian follicle populations were monitored via transrectal ultrasound for 4 d. Four days after estrus, the dominant follicle was aspirated and heifers were treated with FSH-P to induce superovulation. Ovulation rate was determined at ovariectomy 5 d after the superovulatory estrus (experimental Days 35 to 45). Both soybean oil and animal tallow diets increased (P < 0.05) the number of medium-sized follicles and increased (P < 0.02) serum concentrations of GH relative to the control diet. The soybean oil diet also increased (P < 0.001) serum concentrations of insulin on Days 14, 28, and 5 d after the superovulatory estrus. However, the number of ovulations following FSH treatment did not differ due to diet. Procedures employed in the current study were ineffective in recruiting the increased number of medium-sized follicles into the superovulatory pool.     

Exogenous progesterone and progestins as used in estrous synchrony regimens do not mimic the corpus luteum in regulation of luteinizing hormone and 17 beta-estradiol in circulation of cows.

Our working hypothesis was that the low concentrations of progesterone (P4) and synthetic progestins administered in hormonal regimens to control estrous cycles of cows would have similar effects on secretion of LH and 17 beta-estradiol (E2). In addition, we hypothesized that concentrations of exogenous P4 typical of the midluteal phase of the estrous cycle and the corpus luteum (CL) would have similar effects on LH and E2, and the effects would be different from those of synthetic progestins and low concentrations of P4. Cows (n = 29) were randomly assigned to one of five treatment groups: 1) one Progesterone Releasing Intravaginal Device (1PRID; n = 6); 2) two PRIDs (2PRID; n = 6); 3) norgestomet, as in Syncro-Mate-B regimen (SMB; n = 6); 4) melengestrol acetate (MGA; 0.5 mg/day; n = 5); and 5) control (CONT; n = 6). Treatments were administered for 9 days (Day 0 = initiation of treatment). All cows from 1PRID, 2PRID, SMB, and MGA groups were injected with prostaglandin F2 alpha (PGF2 alpha) on Days 2 and 5 of the treatment period to regress CL. Cows in the 1PRID and SMB groups were also administered exogenous estrogen according to the respective estrous synchronization protocol for these products. Daily blood samples were collected from Day 0 to 35 to determine concentrations of P4. On Day 8, blood samples were collected at 15-min intervals for 24 h to determine pattern of LH secretion. On Day 9, all treatments ceased and cows in the CONT group received injections of PGF2 alpha.(ABSTRACT TRUNCATED AT 250 WORDS)     

Changes in ovarian oxytocin secretion as an indicator of corpus luteum response to prostaglandin F(2alpha) treatment in cattle

Exogenous prostaglandin F(2alpha) (PGF(2alpha)) rapidly increases ovarian oxytocin (OT) release and decreases progesterone (P4) secretion in cattle. Hence, the measurement of OT secretion (the area under the curve and the height of the peak) after different doses of Oestrophan - PGF(2alpha) analogue (aPGF(2alpha)) on Days 12 and 18 of the estrous cycle (estrus = day 0), could be a suitable indicator of corpus luteum (CL) sensitivity to PGF(2alpha) treatment. Mature heifers (n = 36) were used in this study. Blood samples were collected from the jugular vein for the estimation of OT, P4 and 13, 14-dihydro-15-keto-prostaglandin F(2alpha) (PGFM). In Experiment 1, different doses of aPGF(2alpha) (400, 300, 200 and 100 microg) given on Day 12 of the estrous cycle (n = 8) shortened (P < 0.05) the cycle duration (15.2 +/- 0.6 d) compared with that of the control (21.7 +/- 0.4 d). Successive heifers were also treated on Day 12 with 200 (n = 2), 100 (n = 2), 75 (n = 2) or 50 microg aPGF(2alpha) (n = 2). Only the 50 microg aPGF(2alpha) dose did not cause CL regression, although it increased OT concentrations to levels comparable to those observed during spontaneous luteolysis (50 to 70 pg/ml). In Experiment 2, on Day 18 of the cycle heifers (n = 8) were treated with 50, 40, 30 and 20 microg aPGF(2alpha). There was a dose-dependent effect of aPGF(2alpha) on OT secretion on Day 18 of the estrous cycle (r = 0.77; P < 0.05). In Experiment 3, an injection of 500 microg aPGF(2alpha) on Day 12 (n = 4) and 50 microg aPGF(2alpha) on Day 18 (n = 4) caused a similar (P > 0.05) increase in the OT concentration (288.5 +/- 23.0 and 261.5 +/- 34.7 pg/ml, respectively). Thus the effect of the same dose of aPGF(2alpha) (50 microg) on OT secretion was different on Days 12 and 18 of the cycle. To evoke similar OT secretion on Days 12 and 18 the dose of aPGF(2alpha) on Day 18 could be reduced 10-fold, confirming that CL sensitivity to PGF(2alpha) appears to increase in the late luteal phase.     

Luteinizing hormone receptors and progesterone content in porcine corpora lutea after prostaglandin F2 alpha

The effect of prostaglandin F2 alpha (PGF2 alpha) on luteinizing hormone (LH) receptors, weight and progesterone content of corpora lutea (CL), and serum progesterone concentrations was studied in gilts. Fifteen gilts were hysterectomized between Days 9 to 11 of the estrous cycle. Twelve gilts were injected i.m. with 10 mg of PGF2 alpha and 3 with saline on Day 20. Ovaries were surgically removed from each of 3 gilts at 4, 8, 12 and 24 h following PGF2 alpha treatment and from the 3 control gilts 12 h following saline injection. Jugular blood samples for progesterone analysis were collected from all gilts at 0, 2 and 4 h following treatment and at 8, 12 and 24 h for gilts from which ovaries were removed at 8, 12 and 24 h, respectively. Mean serum progesterone and CL progesterone concentrations decreased within 4 h after PGF2 alpha treatment (P less than 0.05) and remained low through 24 h after treatment. The number of unoccupied LH receptors decreased by 4 h (P less than 0.05) and this trend continued through 24 h. There were no differences in luteal weight or affinity of unoccupied LH receptors of luteal tissue at 4, 8 12 and 24 h after PGF2 alpha when compared to luteal tissue from controls. These data indicate that during PGF2 alpha-induced luteolysis in the pig, luteal progesterone, serum progesterone concentrations and the number of LH receptors decrease simultaneously.     

Effects of tumor necrosis factor-alpha on secretion of prostaglandins E2 and F2alpha in bovine endometrium throughout the estrous cycle

To determine the physiological significance of tumor necrosis factor-alpha (TNFalpha) in the regulation of endometrial prostaglandin (PG) release in cattle, we investigated the effects of TNFalpha on the secretion of PGE2 and PGF2alpha by bovine endometrium during the estrous cycle. Bovine uteri were classified into six stages (estrus: Day 0, early luteal 1: Days 2 to 3, early luteal 11: Days 5 to 6, mid-luteal: Days 8 to 12, late luteal: Days 15 to 17 and follicular: Days 19 to 21). After 1 h of pre-incubation, endometrial tissues (20 to 30 mg) were exposed to 0 or 0.6 nM TNFalpha for 4 h. The PGE2 concentrations in the medium were higher in the luteal stages than in the follicular stage and in estrus. In contrast, PGF2alpha concentrations were higher in the follicular stage and in estrus than in the luteal stages. The ratio of the basal concentrations of PGE2 and PGF2alpha (PGE2/PGF2alpha ratio) was higher in the luteal stages than in the follicular stage and in estrus. Although TNFalpha stimulated both PGE2 and PGF2alpha secretion during the entire period of the estrous cycle, the level of stimulation of TNFalpha on PGE2 output by the bovine endometrium does not show the same cyclical changes as that shown on PGF2alpha output. The stimulation of TNFalpha resulted in a decrease in the PGE2/PGF2alpha ratio only in the late luteal stage. Furthermore, TNFalpha stimulated PGE2 secretion in stromal, but not epithelial cells. The overall results suggest that TNFalpha is a potent regulator of endometrial PGE2 secretion as well as PGF2alpha secretion during the entire period of estrous cycle, and that TNFalpha plays different roles in the regulation of secretory function of bovine endometrium at different phases of the estrous cycle.     

Estrus induction with prostaglandin F2alpha, cloprostenol or fenprostalene during the normal estrous cycle, superovulation and after embryo collection

Holstein heifers used as embryo donors were treated with three luteolytic agents (PGF2alpha, cloprostenol, fenprostalene) during the normal estrous cycle, superovulation or after embryo collection to determine the interval from treatment to estrus. A similar return-to-estrus interval was observed for each luteolytic agent among the three groups of heifers. Nevertheless, after embryo collection, fenprostalene had a tendency to induce the longest delays (p = 0.08). This tendency is supported by a higher proportion of delayed luteolysis and more heifers showing estrus later than 11 d post treatment. Also, during normal estrous cycles, 5/10 and 0/8 fenprostalene- and cloprostenol-treated heifers, respectively, showed progesterone concentrations higher than 1 ng/mL 48 h after treatment. Regardless of the luteolytic agent used, estrus was induced earlier (P < 0.005) during superovulation than when heifers were treated between Days 9 to 16 of the normal estrous cycle or after embryo collection. However, the return-to-estrus interval was similar between heifers treated during superovulation and those treated between Days 6 to 8 of the normal estrous cycle. After embryo collection, intervals before the return to estrus increased with the number of Corpora lutea (CL) palpated except in the nonresponding group (0 to 1 CL), which returned to estrus later than the low responding group (2 to 4 CL).     

Prostaglandin F2 alpha, oxytocin and progesterone secretion by bovine luteal cells at several stages of luteal development: effects of oxytocin, luteinizing hormone, prostaglandin F2 alpha and estradiol-17 beta

Bovine luteal cells from Days 4, 8, 14 and 18 of the estrous cycle were incubated for 2 h (1 x 10(5) cells/ml) in serum-free media with one or a combination of treatments [control (no hormone), prostaglandin F2 alpha (PGF), oxytocin (OT), estradiol-17 beta (E) or luteinizing hormone (LH)]. Luteal cell conditioned media were then assayed by RIA for progesterone (P), PGF, and OT. Basal secretion of PGF on Days 4, 8, 14 and 18 was 173.8 +/- 66.2, 111.1 +/- 37.8, 57.7 +/- 15.4 and 124.3 +/- 29.9 pg/ml, respectively. Basal release of OT and P was greater on Day 4 (P less than 0.01) than on Day 8, 14 and 18 (OT: 17.5 +/- 2.6 versus 5.6 +/- 0.7, 6.0 +/- 1.4 and 3.1 +/- 0.4 pg/ml; P: 138.9 +/- 19.5 versus 23.2 +/- 7.5, 35.4 +/- 6.5 and 43.6 +/- 8.1 ng/ml, respectively). Oxytocin increased (P less than 0.01) PGF release by luteal cells compared with control cultures irrespective of day of estrous cycle. Estradiol-17 beta stimulated (P less than 0.05) PGF secretion on Days 8, 14 and 18, and LH increased (P less than 0.01) PGF production only on Day 14. Prostaglandin F2 alpha, E and LH had no effect on OT release by luteal cells from any day. Luteinizing hormone alone or in combination with PGF, OT or E increased (P less than 0.01) P secretion by cells from Days 8, 14 and 18. However on Day 8, a combination of PGF + OT and PGF + E decreased (P less than 0.05) LH-stimulated P secretion. These data demonstrate that OT stimulates PGF secretion by bovine luteal cells in vitro. In addition, LH and E also stimulate PGF release but effects may vary with stage of estrous cycle.     

Effect of continuous infusion of oxytocin on length of the oestrous cycle and luteolysis in cattle

In Exp. I oxytocin (60 micrograms/100 kg/day) was infused into the jugular vein of 3 heifers on Days 14-22, 15-18 and 16-19 of the oestrous cycle respectively. In Exp. II 5 heifers were infused with 12 micrograms oxytocin/100 kg/day from Day 15 of the oestrous cycle until clear signs of oestrus. Blood samples were taken from the contralateral jugular vein at 2-h intervals from the start of the infusion. The oestrous cycle before and after treatment served as the controls for each animal. Blood samples were taken less frequently during the control cycles. In Exp. III 3 heifers were infused with 12 micrograms oxytocin/100 kg/day for 50 h before expected oestrus and slaughtered 30-40 min after the end of infusion for determination of oxytocin receptor amounts in the endometrium. Three other heifers slaughtered at the same days of the cycle served as controls. Peripheral concentrations of oxytocin during infusion ranged between 155 and 641 pg/ml in Exp. I and 18 and 25 pg/ml in Exp. II. In 4 our of 8 heifers of Exps I and II, one high pulse of 15-keto-13,14-dihydro-prostaglandin F-2 alpha (PGFM) appeared soon after the start of oxytocin infusion followed by some irregular pulses. The first PGFM pulse was accompanied by a transient (10-14 h) decrease of blood progesterone concentration. High regular pulses of PGFM in all heifers examined were measured between Days 17 and 19 during spontaneous luteolysis. No change in length of the oestrous cycle or secretion patterns of progesterone, PGFM and LH was observed.(ABSTRACT TRUNCATED AT 250 WORDS)     

Premature prostaglandin F2alpha secretion causes luteal regression in GnRH-induced short estrous cycles in cyclic dairy heifers

This study aimed to confirm that the luteolysis in normal-cycling dairy heifers seen during short estrous cycles induced with cloprostenol (Clp) and GnRH administered 24h apart is caused by a premature release of prostaglandin F(2alpha) (PGF(2alpha)). A further aim was to study the PGF(2alpha) release pattern more closely to determine whether it resembles the spontaneous release occurring during normal regression of the corpus luteum (CL) or whether PGF(2alpha) is continuously secreted after the induced ovulations, leading to short estrous cycles. Twenty-four Ayrshire heifers were allotted to four equally sized groups. After estrus synchronization with 0.5mg of Clp, a new luteolysis was induced with 0.5mg of Clp on Day 6 (groups T-d6 and C-d6) or Day 7 (groups T-d7 and C-d7) after ovulation. Gonadorelin (0.1mg i.m.) was given to groups T-d6 and T-d7 to induce premature ovulation 24h later. Groups C-d6 and C-d7 served as controls. Ovaries were examined daily by transrectal ultrasonography, while blood samples (for progesterone and 15-ketodihydro-PGF(2alpha) analyses) were obtained via a jugular catheter every 3h, starting from the second Clp treatment and continuing for 9 days postovulation. Unresponsiveness to Clp or anovulation resulted in 4 C-d6 heifers being excluded. Four heifers in group T-d6 and three in group T-d7 had a short estrous cycle of 8-12 days, while all others had a cycle of normal length. Significant elevations in 15-ketodihydro-PGF(2alpha) concentrations with recurrent high peaks coincided with a decrease in progesterone concentration and were detected in all heifers that showed a short estrous cycle, but not in any heifers with normal estrous cycles in groups T and C. In conclusion, a premature release of PGF(2alpha), which closely resembles its release during spontaneous luteolysis, causes luteal regression in these short cycles.     

Luteinizing hormone and progesterone concentrations in serum of heifers administered a short half-life prostaglandin (PGF(2alpha)) or long half-life prostaglandin analogue (fenprostalene) on days six or eleven of the estrous cycle

Two trials were conducted to measure the progesterone (P(4)) decline and luteinizing hormone (LH) surge in serum subsequent to administration of a short half-life (short t (1 2 )) prostaglandin (PGF(2alpha)) or a long half-life (long t (1 2 )) prostaglandin analogue (fenprostalene) on Days 6 or 11 of the estrous cycle. Twenty-five crossbred Shorthorn and five Hereford heifers with a mean weight of 331.4 +/- 29.8 kg were used in both trials. The heifers were randomly allotted to receive either a short t (1 2 ) or long t (1 2 ) prostaglandin treatment on Day 6 or 11 of the estrous cycle. A crossover design for the main effect, treatment (type of prostaglandin), was conducted. Heifers that received PGF(2alpha) in Trial I were given fenprostalene in Trial II and vice versa. Stage of the estrous cycle (day) was the same for each heifer in both trials. Stage of estrous cycle was standardized to either Day 6 or 11 by administering Syncro-Mate B (SMB). Blood was collected every hour for 80 h post injection to quantify LH and P(4) concentrations. There were no significant differences (P > 0.05) between the short t (1 2 ) or long t (1 2 ) for either P(4) or LH profiles. In addition, no differences were detected between stages of the estrous cycle for the timing of the preovulatory surge of LH after prostaglandin administration.     

Involvement of gonadotropins in induction of luteolysis in pigs

In our previous study we have demonstrated that treatment of endometrial explants with LH increased 13,14-dihydro-15-ketoprostaglandin F(2alpha) (PGFM) accumulation in pigs. This was particularly visible on Days 14-16 of the estrous cycle. Action of gonadotropin in porcine endometrium appears to be mediated by LH/hCG receptors whose number is dependent on the day of the estrous cycle. In the current study i.v. infusion (1 hour) of hCG (200 IU) performed on Days 10 (n=4) and 12-14 (n=4) of the porcine estrous cycle did not affect plasma PGFM (ng/ml+/-SEM) concentrations. In contrast, administration of hCG on Days 15-17 produced, depending on plasma PGFM level before the infusion period, three different types of response: I. plasma PGFM surge of amplitude 0.62+/-0.15 was observed when the mean basal pre-infusion PGFM plasma level was 0.23+/-0.05 (n=6 gilts); II. the delayed PGFM surge of amplitude 0.62+/-0.15 was determined when basal pre-infusion PGFM level was 0.80+/-0.20 (n=6); and III. lack of PGFM response to hCG was found when basal pre-infusion PGFM level was 1.09+/-0.61 (n=6). Concentrations of plasma PGFM before and after saline infusion did not differ on Days 12-14 and 16 of the estrous cycle. In the next experiment blood samples were collected every 1 hour on Days 12-19 of the estrous cycle to determine concentrations of LH, PGFM and progesterone in four gilts. In particular gilts, plasma peaks of LH closely preceded surges of PGFM in 72.7, 84.6, 75.0 and 66.6 percent, respectively. The highest PGFM surges followed a decline in plasma progesterone concentration. We conclude that the increased PGF(2alpha) metabolite production after hCG infusion during the late luteal phase of the estrous cycle as well as the relationship between plasma LH and PGFM peaks suggest the LH involvement in the elevation of endometrial PGF(2alpha) secretion in pigs, and, in consequence, induction of luteolysis.     

Response to GnRH on day 6 of the estrous cycle is diminished as the percentage of Bos indicus breeding increases in Angus, Brangus, and Brahman x Angus heifers

Angus (n=6), Brangus (5/8 Angus x 3/8 Brahman, n=6), and Brahman x Angus (3/8 Angus x 5/8 Brahman, n=6) heifers exhibiting estrous cycles at regular intervals were used to determine if the percentage of Bos indicus breeding influenced the secretory patterns of LH in response to a GnRH treatment on Day 6 of the estrous cycle. Heifers were pre-synchronized with a two-injection PGF(2 alpha) protocol (25 mg i.m. Day -14 and 12.5 mg i.m. Day -3 and -2 of experiment). Heifers received 100 microg GnRH i.m. on Day 6 of the subsequent estrous cycle. Blood samples were collected at -60, -30, and -1 min before GnRH and 15, 30, 60, 90, 120, 150, 180, 240, 300, 360, 420, and 480 min after GnRH to determine concentrations of serum LH. Estradiol concentrations were determined at -60, -30, and -1 min before GnRH. On Day 6 and 8, ovaries were examined by ultrasonography to determine if ovulation occurred. On Day 13, heifers received 25 mg PGF(2 alpha) i.m. and blood samples were collected daily until either the expression of estrus or Day 20 for heifers not exhibiting estrus to determine progesterone concentrations. There was no effect (P>0.10) of breed on ovulation rate to GnRH as well as size of the largest follicle, mean estradiol, and mean corpus luteum volume at GnRH. Mean LH was greater (P<0.05) for Angus (7.0+/-0.8 ng/mL) compared to Brangus (4.6+/-0.8 ng/mL) and Brahman x Angus (2.9+/-0.8 ng/mL), which were similar (P>0.10). Mean LH peak-height was similar (P>0.10) for Brangus (13.9+/-3.4 ng/mL) compared to Angus (21.9+/-3.4 ng/mL) and Brahman x Angus (8.0+/-3.4 ng/mL), but was greater (P<0.05) for Angus compared to Brahman x Angus. Interval from GnRH to LH peak was similar (P>0.10) between breeds. As the percentage of Bos indicus breeding increased the amount of LH released in response to GnRH on Day 6 of the estrous cycle decreased.     

Comparison of controlled internal drug release device and melengesterol acetate as progestin sources in an estrous synchronization protocol for beef heifers

The objectives of this experiment were to compare estrous synchronization responses and AI pregnancy rates of beef heifers using protocols that included either CIDR or MGA as the progestin source. The hypotheses tested were that: (1) estrous synchronization responses after (a) progestin removal, and (b) PGF(2alpha); and, (2) AI pregnancy rates, do not differ between heifers synchronized with either progestin source. At the start of the experiment (Day 0) in both years, heifers were assigned randomly to receive, MGA supplement for 14 days (MGA-treated; n=79) or CIDR for 14 days (CIDR-treated; n=77). On Day 14 progestin was removed and heifers were observed for estrus up to and after PGF(2alpha) on Days 31 and 33 for CIDR-treated and MGA-treated heifers, respectively. Heifers that exhibited estrus within 60h after PGF(2alpha) were inseminated by AI 12h later; the remaining heifers were inseminated at 72h after PGF(2alpha) and given GnRH (100mug). More (P<0.05) CIDR-treated heifers exhibited estrus within 120h after progestin removal than MGA-treated heifers. Intervals to estrus after progestin removal were shorter (P<0.05) for CIDR-treated heifers than MGA-treated heifers. More (P<0.05) CIDR-treated heifers exhibited estrus and were inseminated within 60h after PGF(2alpha) than MGA-treated heifers. Pregnancy rates did not differ (P>0.10) between MGA-treated (66%) and CIDR-treated (62%) heifers. In conclusion, the use of CIDR as a progestin source in a 14-day progestin, PGF(2alpha), and timed AI and GnRH estrous synchronization protocol was as effective as the use of MGA to synchronize estrus and generate AI pregnancies in beef heifers.     

Effect of stage of the estrous cycle on interval to estrus after PGF(2alpha) in beef cattle

Effect of stage of the estrous cycle at the time of prostaglandin F(2alpha) (PGF(2alpha)) injection on subsequent reproductive events in beef females was studied in four trials involving 194 animals. Cycling animals were given two injections of 25 mg PGF(2alpha) 11 days apart or, in some cases, the interval was altered to allow the second injection to fall on a specific day of the cycle. Day of estrous cycle at time of the second injection was determined by estrous detection. Interval from the second PGF(2alpha) injection to the onset of estrus (interval to estrus) was shorter (P<.01) in heifers than in cows. Both cows and heifers injected on days 5 to 9 (early cycle) had a shorter (P<.01) interval to estrus (estrus = day 0) than did those injected on days 10 to 15 (late cycle). Conception rate was lower (P<.05) for early-cycle heifers than for late-cycle heifers inseminated by appointment at 80 hours. There was no significant difference in conception rate of early-or late-cycle heifers or cows inseminated according to estrous detection or early- or late-cycle cows inseminated at 80 hours. Progesterone concentrations in blood samples collected in heifers at 4-hour intervals after the second PGF(2alpha) injection on either day 7 or day 14 declined linearly (P<.05) through 36 hours. Day of the estrous cycle at PGF(2alpha) injection had no effect on rate of progesterone decline, even though heifers injected on day 7 had a shorter (P<.05) interval to estrus. All animals whose cycle length was not affected by the second PGF(2alpha) injection were treated on days 5 through 8 of the cycle, indicating that PGF(2alpha) was less effective in regressing the corpus luteum between days 4 and 9 of the cycle than later in the cycle.     

Effects of progesterone administered to dairy heifers on sensitivity of corpora lutea to PGF(2)alpha and on plasma LH concentration

To determine whether progesterone facilitates PGF(2)alpha-induced luteolysis prior to day 5 of the estrous cycle, 48 Holstein-Friestian heifers were assigned at random to four treatments: 1) 4 ml corn oil/day + 5 ml Tris-HCl buffer (control); 2) 25 mg prostaglandin F(2)alpha (PGF(2)alpha); 3) 100 mg progesterone/day (progesterone); 4) 100 mg progesterone/day + 25 mg PGF(2)alpha (combined treatment). Progesterone was injected subcutaneously daily from estrus (day 0) through day 3. The PGF(2)alpha was injected intramuscularly on day 3. Estrous cycle lengths were decreased by progesterone: 20.2 +/- 0.56, 19.2 +/- 0.31 (control and PGF(2)alpha); 13.2 +/- 1.40, and 11.7 +/- 1.27 (progesterone and combined). The combination of progesterone and PGF(2)alpha did not shorten the cycle any more than did progesterone alone (interaction, P>0.05). PGF(2)alpha treatment reduced progesterone concentrations on day 6 (P<0.05) and both progesterone and PGF(2)alpha reduced plasma progesterone on day 8 (P<0.01 and P<0.05, respectively). LH was measured in blood samples collected at 10- min intervals for 4 hr on day 4 from three heifers selected at random from each of the four treatment groups. Mean LH concentration for control heifers ranged from 0.35 to 0.63 ng/ml (overall mean, 0.49 ng/ml) and for progesterone-treated heifers ranged from 0.12 to 0.30 ng/ml (overall mean, 0.23 ng/ml). LH concentrations were greater in control heifers (P<0.01). The mean LH pulse rate for control heifers was 2.7 pulses/heifers/4 hr, while that for the progesterone-treated heifers was 1.7 pulses/heifer/4 hr. The mean pulse amplitude for control and progesterone treatments was 0.47 ng/ml and 0.36 ng/ml, respectively. Neither pulse amplitude nor frequency were different between treatment groups.     

Sensitivity of bovine corpora lutea to prostaglandin F2alpha is dependent on progesterone, oxytocin, and prostaglandins.

Prostaglandin (PG) F2alpha that is released from the uterus is essential for spontaneous luteolysis in cattle. Although PGF2alpha and its analogues are extensively used to synchronize the estrous cycle by inducing luteolysis, corpora lutea (CL) at the early stage of the estrous cycle are resistant to the luteolytic effect of PGF2alpha. We examined the sensitivity of bovine CL to PGF2alpha treatment in vitro and determined whether the changes in the response of CL to PGF2alpha are dependent on progesterone (P4), oxytocin (OT), and PGs produced locally. Bovine luteal cells from early (Days 4-5 of the estrous cycle) and mid-cycle CL (Days 8-12 of the estrous cycle) were preexposed for 12 h to a P4 antagonist (onapristone: OP; 10(-4) M), an OT antagonist (atosiban: AT; 10(-6) M), or indomethacin (INDO; 10(-4) M) before stimulation with PGF2alpha. Although OP reduced P4 secretion (p < 0.001) only in early CL, it reduced OT secretion in the cells of both phases examined (p < 0.001). OP also reduced PGF2alpha and PGE2 secretion (p < 0.01) from early CL. However, it stimulated PGF2alpha secretion in mid-cycle luteal cells (p < 0.001). AT reduced P4 secretion in early and mid-cycle CL (p < 0.05). Moreover, PGF2alpha secretion was inhibited (p < 0.05) by AT in early CL. The OT secretion and the intracellular level of free Ca2+ ([Ca2+]i) were measured as indicators of CL sensitivity to PGF2alpha. PGF2alpha had no influence on OT secretion, although [Ca2+]i increased (p < 0.05) in the early CL. However, the effect of PGF2alpha was augmented (p < 0.01) in cells after pretreatment with OP, AT, and INDO in comparison with the controls. In mid-cycle luteal cells, PGF2alpha induced 2-fold increases in OT secretion and [Ca2+]i. However, in contrast to results in early CL, these increases were magnified only by preexposure of the cells to AT (p < 0.05). These results indicate that luteal P4, OT, and PGs are components of an autocrine/paracrine positive feedback cascade in bovine early to mid-cycle CL and may be responsible for the resistance of the early bovine CL to the exogenous PGF2alpha action.     

Influence of day length and endocrine status on luteinizing hormone secretion in intact and ovariectomized adult ferrets

The purpose of this study was to examine the pituitary-ovarian relationship of both estrous and anestrous female ferrets. The endocrine status of the animals was induced by manipulating photoperiod: females in estrus were housed in long days (16L:8D); females in anestrus were housed in short days (8L:16D). For studies of intact animals in both photoperiods, plasma luteinizing hormone (LH) levels were quantified in blood samples collected from adult ferrets at 5-min intervals over a 24-h period. Similar groups of females (estrous and anestrous) were ovariectomized (while remaining in their assigned photoperiods) and blood samples were collected at 5-min intervals for 4-h periods on Days 1, 2, 4, 10, 17, and 35 after ovariectomy. Intact, estrous females exhibited continuously low or undetectable levels of LH with no evidence of episodic secretion. Ovariectomy of these estrous animals resulted in rapid onset (within 24 h) of episodic LH secretion, with pulses occurring in excess of 1 pulse/h. No substantial further change in frequency or amplitude of pulses occurred in these females from 1 to 35 days postovariectomy. In contrast, intact anestrous ferrets exhibited clear episodic LH secretion at a frequency of about 0.4 pulses/h. Removal of ovaries from these females caused no change in LH secretion for 24-48 h, after which LH pulses gradually increased in frequency. By 18 days after ovariectomy, LH patterns were indistinguishable among ovariectomized females in long and short days. These studies suggest a major site of ovarian negative feedback on LH secretion during anestrus is the hypothalamus, whereas the site of the ovarian feedback in estrous females is not yet evident.     

Effect of timing of prostaglandin PGF 2 alpha injection subsequent to embryo collection on the resumption of normal follicular development following superovulatory treatment in cattle

Nonlactating Holstein and Jersey cows (n = 24) were superovulated and ovarian follicular development was monitored by transrectal ultrasound during the period after embryo recovery. Luteolysis was induced by two injections of prostaglandin F(2)alpha (PGF; 25 mg Lutalyse; 12-h interval) at specific times after superovulatory induced estrus (Treatment 1, Day 9; Treatment 2, Day 12; Treatment 3, Day 17; Treatment 4, Day 25; superovulatory estrus = Day 0 of Cycle 1). Follicular development was monitored during Cycle 1 before and after PGF injection and continued through the ensuing estrous cycle (Cycle 2). Superovulation led to more than one embryo collected in 14 cows (mean = 8.71 embryos: positive superovulatory response [PSR] cows), while 10 cows were not successfully superovulated (mean = 0.1 embryo; negative superovulatory response [NSR] cows). These cows differed in terms of number of unovulated follicles detected at embryo collection (4.21 vs 17.2, PSR vs NSR) and plasma progesterone during the superovulatory estrous cycle (32.3 ng/ml PSR vs 8.6 ng/ml NSR). Follicular development during Cycle 1 started sooner in NSR than in PSR cows (day by class by response P<0.03) and was initiated on Days 11 to 12 in NSR cows and on Days 19 to 20 in PSR cows. Interval to estrus after PGF averaged 6.3 d. Cows having short intervals to estrus had follicles at the time of PGF injection. Treatment influenced the length of Cycle 1, but it did not affect the interval to estrus after PGF, the length of Cycle 2, or follicular development during Cycle 2. The results indicate that 1) the timing of PGF injection after embryo collection does not influence subsequent follicular populations, 2) elongated estrous cycles and intervals to estrus after PGF in superovulated cattle are a function of decreased follicular activity, and 3) the presence of numerous corpora lutea and not the superovulatory treatment, per se, seem to attenuate follicular growth.