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Ability to select service sires that minimize partial or complete losses of pregnancy could have major economic impacts in sheep production systems. This study tested the null hypothesis that survival of potential progeny did not vary with breed type of service sire or among individual rams. Data included 980 ewes on 10 farms; each ewe was pregnant to 1 of 67 rams of 12 breeds. Number of conceptuses was estimated once during pregnancy by ultrasonography, either transrectal (embryos) or transabdominal (fetuses), and was compared with number of lambs born to estimate losses. Data were examined first for number of lambs born and second for documented losses. Individual service sires affected number born (P < 0.001), which varied from 0.70 to 2.45 lambs per pregnant ewe. The main effects of breed type on lambs born were not significant, but breed types of both service sires (P < 0.0002) and ewes (P < 0.001) interacted with diagnosed number of conceptuses. Lambs born varied with ewe age (P < 0.0001) and among farms (P < 0.0001), and statistically, farms interacted with number of diagnosed conceptuses (P < 0.0001); season had no effect. In documented losses, there were both main effects of individual service sire and a service sire × number of diagnosed embryos interaction (P < 0.005). Thus, ewes bred to some rams were more apt to lose single pregnancies, whereas ewes bred to other rams were more apt to lose 1 or more embryos or fetuses from multiple pregnancies. Breed type of service sire affected (P < 0.05) prenatal death. Complete losses of single conceptuses tended to be greater in ewes bred to black-faced or hair-type rams (service sire breed type × number of diagnosed conceptuses; P < 0.09). Breed type of ewes also varied in incidence of complete losses (P < 0.05); hair-type ewes (46%) lost more (P < 0.02) documented conceptuses from examination to birth than black-faced (27%), white-faced (20%), or dairy-type (25%) ewes. Greater losses of singles than of multiples occurred in black-faced (37% vs. 18%) and hair-type (64% vs. 27%) ewes than in other breeds (ewe breed type × number of conceptuses; P < 0.03) per ewe. Surprisingly, purebred conceptuses were lost less often (24%) than crossbreds (36.4%; P < 0.002). Selection of rams based on records of prenatal losses in ewes they serviced may be a method to decrease embryonic and fetal wastage. However, further study to determine repeatability of differences among service sires from year to year will be required.

Three experiments were designed to examine the mechanisms that govern prostaglandin (PGF2alpha)-induced regression of the sheep corpus luteum. Evidence is presented supporting the involvement of endothelin 1 (EDN1) in PGF2alpha-induced luteolysis. Experiment 1 measured effects of PGF2alpha when actions of EDN1 were blocked by sustained administration of a type-A endothelin (EDNRA) or type-B endothelin (EDNRB) antagonist in vivo. Experiment 2 examined antisteroidogenic actions of PGF2alpha and EDN1 in the presence of an EDNRA or EDNRB antagonist in Day-8 luteal minces. In experiment 3, luteal cellular expression of EDNRA and EDNRB was determined immunohistochemically. Relative gene expression of EDNRA and EDNRB receptors was examined by real-time RT-PCR in Day-8 sheep corpora lutea. EDNRA, but not EDNRB, participated in antisteroidogenic actions of EDN1. During the first 12 h after PGF2alpha-induced luteolysis, EDNRA antagonist did not prevent a decline in serum progesterone concentrations. Early actions of PGF2alpha are either direct or mediated by something other than EDN1. However, beyond 12 h after PGF2alpha, serum progesterone concentrations increased in EDNRA antagonist-treated animals until they were the same as saline-treated controls, whereas an EDNRB antagonist had no effect in vivo or in vitro. The EDNRA antagonist negated the antisteroidogenic actions of EDN1 but only partially abolished the actions of PGF2alpha in vitro. In contrast, the EDNRB antagonist was ineffective in abolishing antisteroidogenic actions of EDN1 and PGF2alpha. Whereas real-time RT-PCR demonstrated high expression of EDNRA and low expression of EDNRB, immunohistochemically, only EDNRA was located in small steroidogenic, endothelial, and smooth muscle cells. In summary, studies in ovine corpora lutea provided strong evidence that: 1) EDNRA, but not EDNRB, mediates antisteroidogenic actions of EDN1, 2) actions of PGF2alpha are both independent of and dependent upon mediation by EDN1, and 3) small steroidogenic cells are targets for antisteroidogenic effects of EDN1. Furthermore, the results from experiment 1 suggest that the intermediary role of EDN1 may be more important in later stages of luteal regression.

The objective was to determine if transrectal ultrasonography for determination of pregnancy in restrained ewes increases embryonic/fetal death or loss of pregnancy. Ten flocks (N=873 ewes) bred in either the estrous or anestrous season were randomized, into control (C) or examined (E) groups within flock. Examined ewes were placed in a tilting squeeze chute and scanned by one of three operators for pregnancy by transrectal ultrasonography once between Days 25 and 100 post-breeding. Control ewes were not subjected to handling in the squeeze chute or pregnancy diagnosis. Two operators counted embryos in E ewes in six flocks. There were no differences between E and C ewes in percent ewes lambing or lambing rate on a per flock basis. Prolificacy per flock was greater in E ewes (P=0.05; 1.53 versus 1.60, C and E, respectively) than in C ewes. Eighty-five percent of exposed ewes lambed in the estrous season, while only 62% of ewes lambed in the anestrous season (P=0.05). Overall prolificacy did not differ with season (1.60 versus 1.53), and there was no season by treatment interaction for any variable tested. Losses averaged 0.02 per E ewe exposed in the estrous season and 0.20 in the anestrous season (P<0.05). Examined ewes in this study had similar pregnancy and lambing rates to C ewes in the same flocks and prolificacy was slightly greater. Based on these data, the combination of transrectal ultrasonography with restraint is safe for pregnancy diagnosis in ewes.

During maternal recognition of pregnancy, the conceptus stimulates endometrial secretion of PGF2alpha and PGE2. However, PGF2alpha is less effective in causing luteal regression in pregnant than in non-pregnant ewes. Experiments were conducted to elucidate mechanisms for reduced luteal sensitivity to PGF2alpha during maternal recognition of pregnancy. Corpora lutea (CL) were collected from pregnant and non-pregnant ewes 0, 4, or 12h following treatment with PGF2alpha on day 12 after estrus. Luteal PTGHS2 mRNA did not differ due to PGF2alpha or pregnancy status. Luteal PTGES mRNA was reduced in both pregnant and non-pregnant ewes after PGF2alpha treatment; while, luteal PTGFS mRNA was reduced 4h after PGF2alpha in pregnant, but not non-pregnant ewes. The result was a greater ratio of PTGES to PTGFS mRNA in pregnant ewes. Luteal mRNA for HPGD did not differ between pregnant and non-pregnant ewes on day 12. Luteal END1 mRNA was reduced in pregnant as compared to non-pregnant ewes prior to PGF2alpha challenge. Luteal END1 mRNA was increased after PGF2alpha in pregnant and non-pregnant ewes; however, ECE1 mRNA was reduced 4h after PGF2alpha in pregnant, but not non-pregnant ewes. The in vitro conversion of PGF2alpha to PGFM was greater in CL of pregnant than non-pregnant ewes at day 14. Luteal conversion of PGF2alpha to PGFM appears to be regulated post-transcriptionally. During maternal recognition of pregnancy, mechanisms of reduced luteal sensitivity to PGF2alpha may include a shift in prostaglandin production to the luteotropin PGE2, a reduction of ECE1 mRNA, and increased catabolism of PGF2alpha.

Three experiments were conducted with a total of 1579 ewes to examine reproductive performance in response to synchronization of oestrus during the breeding season, using controlled internal drug releasing (CIDR-G) inserts in regimens designed to provide high concentrations of circulating progesterone. In experiment 1, treatment with two CIDR-G inserts for 12 days produced conception rate (79%) and prolificacy (1.9) to first service equivalent to breeding at natural oestrus (56% and 2.0, respectively). Pregnancy rates to two service periods were 90 and 79%, respectively. In experiments 2 and 3, progesterone was delivered by a single CIDR-G insert for 5 days in combination with prostaglandin F2alpha (PGF2alpha; 5 mg i.m., twice, 3 h apart) the day before (experiment 2), or at insert removal (experiment 3). The combined treatments improved rates of synchronization of oestrus (p<0.01) by 23 and 20% points, respectively, and pregnancy rates to the first service period by 19 (p<0.05) and 13 (p<0.01) percentage points, respectively, compared to treatment with PGF2alpha alone. It is concluded that the combination of treatment for 5 days with a CIDR-G insert and two injections of 5 mg PGF2alpha, the day before, or the day of insert removal, were effective treatments to obtain high fertility at synchronized oestrus in ewes during the breeding season.

Experiments were performed to determine the effects of components of the GH-IGF axis on conceptus development and postnatal growth in sheep. In Exp. 1, ewes received one of the following treatments: 1) sustained release GH at breeding, 2) sustained release GH at breeding and estradiol-17beta at d 5 and 6, 3) only estradiol-17beta at d 5 and 6, or 4) no treatment. Uteri were flushed on d 7, and flushings were analyzed for content of IGF-I. A single injection of sustained-release bovine GH at breeding increased IGF-I content in uterine luminal flushings compared with control ewes (P < 0.05). Treatment with estradiol-17beta on d 5 and 6 after breeding did not alter IGF-I content compared with control ewes, and it blocked the effect of GH on uterine luminal IGF-I content. In Exp. 2, sustained release GH or no treatment was administered at breeding, and gravid uteri were collected at d 25, 80, or 140 of gestation. On d 80, GH-treated ewes had smaller chorioallantoic weights (P < 0.05) and tended to have more efficient placentae (fetal weight/total placental weight; P = 0.052), with a higher percentage of placental weight as cotyledons (P = 0.068) compared with control ewes. In Exp. 3, ewes were treated with or without sustained release GH at progesterone withdrawal. Lambs from GH-treated ewes were heavier at birth (P < 0.05). Lambs from GH-treated ewes reared as singles, but not lambs reared as multiples, were heavier at 30, 60 (P < 0.05), and 75 d (P = 0.075) of age than lambs from control ewes. In conclusion, ewes treated with sustained-release GH at breeding developed smaller, more efficient placentas, and had larger lambs at birth.

Experiments were performed to test the null hypotheses that embryonic survival is not affected by the presence of regressing corpora lutea in progestogen-supplemented ewes, and that the embryotoxic effects of regressing corpora lutea do not act locally on embryos in the uterine horn adjacent to the regressing corpora lutea. In Expt 1, laparotomies were performed on day 4 after mating, and progestogen supplementation was initiated and continued until pregnancy was diagnosed by ultrasonography on day 25. On day 4 after mating, ewes were lutectomized (n = 17) or sham lutectomized (n = 15), and injected (i.m.) with 5 mg PGF2alpha at 8 h intervals for 2 or 3 days. Controls (n = 14) were sham lutectomized and injected with saline as described above. Pregnancy rates did not differ in ewes treated with PGF2alpha for 2 rather than 3 days. Pregnancy rates were lower in ewes treated with PGF 2a compared with controls (P < 0.01). In ewes treated with PGF2alpha, lutectomy resulted in an increase in pregnancy rates (59%) compared with ewes subjected to sham lutectomy (33%; P < 0.05). In Expt 2, progestogen supplementation was initiated in the morning of day 4 after mating and continued until pregnancy diagnosis on day 25. In the afternoon of day 4, one ovary selected at random was lutectomized in ewes (n = 34) with at least one corpus luteum on each ovary, and the uterine horns were isolated by ligation to impede intraluminal transfer of luteal or uterine products that might initiate embryonic death. On days 5-8 after mating, equal numbers of ewes were injected i.m. with either saline or 5 mg PGF2alpha at 8 h intervals. Pregnancy rates did not differ between isolated uterine horns contralateral and ipsilateral to the regressing corpus luteum; however, pregnancy rates were lower in PGF2alpha-treated ewes than in saline-treated ewes (34 and 77%; P < 0.05). In conclusion, regressing corpora lutea exert an embryotoxic effect; however, there is no evidence that this effect occurs through systemic pathways.

The objectives of this study were to determine whether a new progesterone (P4)-releasing intravaginal insert would induce fertile estrus and whether FSH combined with the insert would increase prolificacy in anestrous ewes introduced to rams. Ewes of mixed breeding on six farms were assigned to four randomized treatments: control (C), n = 73; 12 d P4 (polycapralactone [PCL] insert with 0.82 g P4), (P12), n = 73; 12 d P4 plus i.m. FSH (Folltropin, 55 mg NIH-FSH-P1 equivalent) in propylene glycol, 24 h before insert removal, (P12F), n = 71; and 5 d P4 plus FSH (P5F), n = 77. Growth and ovulation of follicles were observed ultrasonographically in 20 ewes at four farms (five/treatment) at insert removal and 36, 48, 72, and 96 h later. Intact rams (1:15 ewes in multiple-sire groups) were joined at insert removal, and raddle marks were observed every 12 h for 5 d. On d 26 to 30, rams were removed; ewes were examined for pregnancy then and 20 d later. Percentage of ewes marked by rams was greater in P4-treated (66 to 79%) than in C (12%; P < 0.01) ewes and in P5F (79%) than in P12F (66%; P < 0.05). Diameters of largest follicles at insert removal were greater (P < 0.05) in P4-treated (5.5 +/- 0.2) than in C ewes (4.8 +/- 0.2). Progesterone increased numbers of follicles > 3 mm (P < 0.01) or ovulated (P < 0.05; 2.6 +/- 0.6 vs 1.3 +/- 0.6 in C ewes) and FSH increased number of follicles > 3 mm (P < 0.05). In FSH-treated ewes, ovulation rate tended to be greater after treatment with P4 for 5 than for 12 d (P = 0.09, 3.3 +/- 0.6 and 2.2 +/- 0.4, respectively). More P4-treated than C ewes lambed (P < 0.01) to the first (38 to 45 vs 0%) or both (63 to 66 vs 41%) service periods. Prolificacy (first service) did not differ between FSH-treated ewes (P12F + P5F; 1.8 +/- 0.1) and ewes treated with P4 only (P12; 1.6 +/- 0.1). However, FSH increased prolificacy to first service (1.8 +/- 0.1) over prolificacy to second service (C ewes 1.5 +/- 0.1; P < 0.05, and all ewes 1.4 +/- 0.1; P < 0.01). Pregnancy retention did not differ among treatments but was greater (P < 0.01) in ewes that conceived at the first (90.9 +/- 3.7) than at the second (72.5 +/- 3.3) service period. In conclusion, a PCL insert in combination with ram introduction at insert removal was more effective than ram introduction alone to induce synchronized estrus and ovulation and to yield pregnancy after one or two service periods. Treatment with P4 for 5 d was as effective as for 12 d to induce fertile estrus in FSH-treated anestrous ewes.

The objectives were to evaluate, in anestrous ewes, the effectiveness of a CIDR-G device (0.3 g progesterone) administered for 5 d to induce estrus; and FSH (Folltropin; 55 mg NIH-FSH-P1 equivalent) in saline:propylene glycol (1:4) 24 h before insert removal (Day 0), to increase ovulation rate and prolificacy. Ewes of mixed breeding were assigned at random to 3 treatments: control (C; n = 125), 5 d progesterone (P5; n = 257) and 5 d progesterone plus FSH (P5F; n = 271). Intact rams were joined at insert removal and ewes were observed every 24 h for 3 d. On Day 14, the ovulation rates of all ewes detected in estrus in the treated groups were determined using transrectal ultrasonography. Rams were removed on Day 26 to 31. Ewes were examined for pregnancy then, and again 20 to 25 d later to detect ewes that conceived to the second service period. Percentage of ewes marked by rams was higher in progesterone-treated (77%) than in C (20%; P < 0.01), but did not differ between P5 and P5F. The ovulation rate (1.95+/-0.04) did not differ due to FSH. Conception (68%) and pregnancy (52%) rates were higher in progesterone-treated (P < 0.01) than in C (0%) ewes. Estrous response varied quadratically with time after ram introduction, and the conception rate varied quadratically with the time of observation of onset of estrus. Over two service periods more progesterone-treated than C ewes lambed (65 vs 45%; P < 0.01). Lambs born per ewe exposed (0.7+/-0.1, 1.0+/-0.1, and 1.1+/-0.1 for C, P5 and P5F, respectively) was increased by progesterone (P < 0.05). Litter size to the first service period (1.59+/-0.04) and overall (1.54+/-0.03) did not differ among treatment groups. FSH-treated ewes tended to have more lambs (1.67+/-0.1) than did ewes receiving progesterone alone (1.5+/-0.1; P = 0.06) and than did ewes lambing to the second service period (1.5+/-0.1; P = 0.06). In summary, a 5-d progesterone pre-treatment of anestrous ewes induced estrous cycles and increased the pregnancy rates. A single injection of FSH only tended to increase litter size.

Growth and regression of ovarian follicles with antral diameters > or = 2 mm were characterized during 15 estrous cycles by daily transrectal ultrasonography (7.5 MHz probe) in 9 ewes of Merino del Pais, a consistently monovular Spanish breed. Mean interovulatory interval was 17.5 +/- 0.5 days and ovulation rate was 1 in all ewes; of 60 to 116 follicles, > or = 2 mm observed during the entire estrous cycle, 13.0 +/- 1.2 reached a maximum diameter > or = 4 mm and 7.9 +/- 0.6 different follicles became the largest follicle in the animal at some point during the cycle. An average of 4.5 new follicles per ewe were detected each day, with no significant effect of day of cycle. Appearance of new follicles that grew to > or = 4 mm tended to differ during the first 8 days of the cycle, being highest on day 3 and lowest on day 6 (P < 0.10), but did not vary significantly during the last 6 days. Growth of new follicles from the day of detection to the next day differed between, but not within, periods, averaging 1.4 +/- 0.3 mm of the first 8 days of the cycle and 1.8 +/- 0.5 mm from day -6 through -1 (P < 0.05). Total number of follicles > or = 2 mm per ovary on days 1 through 8 varied with the interaction of ovary by day, being more variable in the non-CL ovary. During the last 7 days, a linear decline in total follicles was coupled with a linear increase in number of large follicles (P < 0.05). Differences in the size between the largest and second largest follicles were greater on days 5 through 8 than on days 1 through 4, did not differ with day of cycle on days -6 through -1, then increased on the last day from 1.5 mm to 2.9 mm (P < 0.001). In conclusion, the monovular Merino del Pais ewe showed a more rapid growth and turnover of ovarian follicles than other breeds studied, but identified 3-mm follicles did not emerge in other than a random distribution. There was little evidence of dominance until the ovulatory follicle had been identified.

The effects of progesterone (P4) on follicular growth and fertility in ewes were examined. In Experiment 1, 22 ewes received either one or three packets of P4 (5 g/packet) or an empty packet subcutaneously (sc) from Days 5 to 15 of the estrous cycle (estrus = Day 0). On Day 6, P4-treated ewes received 12.5 mg of prostaglandin F2 alpha. Follicles > or = 3 mm in diameter were observed via transrectal ultrasonography daily from Day 4 through estrus, corpora lutea (CL) were observed 5 to 7 d after estrus. Ewes with low (LOW; < or = 1 ng/ml; n = 5), intermediate (MED; >1 and <2 ng/ml; n = 10), or normal (NOR; > or =2 ng/ml; n = 7) P4 in jugular plasma on Days 7 through 15 differed in follicular development. The largest follicle at estrus was larger in ewes with LOW vs. MED and NOR P4 (7.8 +/- 0.3 vs. 6.9 +/- 0.2 mm; P < 0.05). Treatments differed in proportions of multiple-ovulating ewes, in which the oldest ovulatory follicle was first observed before Day 10 (LOW: 3 of 3, MED: 6 of 10, NOR: 0 of 5, respectively; P < 0.05). Estradiol was higher early in the treatment period in LOW ewes than in MED and NOR ewes (day x treatment; P < 0.05). In Experiment 2, ewes received 5 mg of P4 in corn oil (low progesterone [LP]; n = 51) or 2 ml of corn oil (CON; n = 49) sc every 12 hr on Days 6 through 14 of the estrous cycle before mating. LP ewes received 15 mg of prostaglandin F2 alpha on Day 6. Mean serum P4 on Days 7 through 15 was 0.6 +/- 0.1 ng/ml in LP and 1.9 +/- 0.1 ng/ml in CON ewes. Eleven LP and 12 CON ewes were scanned daily from Day 4 through mating, and in all ewes (n = 93), CL were counted 10 d after mating and embryos were counted at 25, 40, and 60 d of gestation. In multiple-ovulating ewes, day of cycle of appearance was earlier for the oldest (Day 6.1 +/- 0.8 vs. 10.4 +/- 0.8) but not second oldest (Day 11.7 +/- 1.0 vs. 12.2 +/- 0.9) ovulatory follicles in LP compared with CON ewes. The conception rate was lower in LP (72%) than in CON ewes (98%; P < 0.01). However, numbers of CL 10 d after mating, and in pregnant ewes, numbers of embryos 25 d after mating and lambs born, did not differ with treatment. In summary, low P4 increased the size of the largest follicles and the age of the oldest ovulatory follicles. Embryos resulting from the ovulation of older and younger follicles in the same ewe did not differ in their ability to survive.

Effects of day of estrous cycle and induced luteal regression on follicular dynamics in ewes were examined. In Exp. 1, ewes received PGF2 alpha (i.m.) on d 5, 8, or 11 after estrus (d 0) or saline on d 14 (n = 27, 27, 31, and 29, respectively). Ewes were laparotomized before treatment and again 48 h after induced estrus to record numbers, sizes, and locations of follicles > or = 2 mm and corpora lutea (CL). The two largest follicles (designated F1 and F2) and any of equal size (F1.5 and F2.5) were marked with India ink in stroma around the follicle. Ewes treated on d 8 ovulated fewer F1 and F1.5 (39%) than ewes treated on d 5 (69%; P < .05); d 11 (51%) and 14 (56%) were intermediate. In Exp. 2, ewes received PGF2 alpha on d 5 or 8 (n = 15/d). Ovaries were examined with transrectal ultrasonography daily from 4 d before PGF2 alpha until ovulation, and on d 5 and 8 after ovulation. Diameters, locations, and ranks (at time of PGF2 alpha) of follicles > or = 2 mm and CL were recorded. Ewes treated on d 8 ovulated fewer F1 and F1.5 (35%) than ewes treated on d 5 (86%; P < .05). Ovulatory follicles at time of PGF2 alpha were larger on d 5 (4.7 +/- .2 mm) than on d 8 (4.0 +/- .2 mm; P < .05).(ABSTRACT TRUNCATED AT 250 WORDS)

This study was conducted to determine the effects of the corpus luteum (CL) of pregnancy and(or) the previously gravid uterine horn (g-UH) on the incidence and site of ovulation and the duration of luteal function in postpartum (PP) ewes. Ewes were laparotomized at d 60 to 80 of gestation and assigned to one of four groups: 1) single g-UH, CL left intact (SI, n = 35); 2) single g-UH, CL removed (SR, n = 30); 3) twin g-UH, CL on both ovaries left intact (TI, n = 21); and 4) twin g-UH, CL removed from one ovary (TR, n = 32). On d 20 PP, the number of follicles on each ovary was determined at laparoscopy, and ewes received (i.m.) 750 IU of hCG. Ovarian structures were determined at laparoscopy on d 24 PP. Blood samples were collected daily from d 20 to 40 PP and assayed for progesterone. more follicles (P < .05) were observed at laparoscopy on d 20 PP in SI/TI than in SR/TR groups (8.4 vs 6.0, respectively; SE = .4). Overall, 52% of the ewes had visible CL 4 d after hCG, and 73% of the ewes had elevated levels of progesterone by 7 d after hCG. The number of g-UH (S vs T) or status of the CL (I vs R) did not affect these responses. For SR ewes with hCG-induced CL on a single ovary, the incidence of CL formation on the ovary adjacent to g-UH was less (P < .05) than that for SI ewes (0 vs 57, respectively). Removal of the CL from S ewes, but not from T ewes, reduced follicular development and ovulation in response to hCG. Perhaps there is a positive systemic effect of the CL of pregnancy on postpartum ovarian function in ewes.

Three experiments were performed to study effects of decreased concentrations of estradiol-17 beta (E2) on lifespan and function of ensuing ovine corpora lutea (CL). In experiment 1, 52 follicles were collected from 10 ewes and placed into individual culture with 0 or .01 microCi 3H-androstenedione (10 ng; 3H-A) and 0, 10(-11), 10(-9), 10(-7), or 10(-5) M of a nonsteroidal aromatase inhibitor, CGS16949A (CGS). Concentrations of E2 secreted into the medium, and synthesis of estrogens as estimated by formation of 3H-water from 3H-A were decreased by 10(-5) and 10(-7) (P < .01), but not 10(-9) or 10(-11) M CGS. In experiment 2, luteolysis was induced in 24 ewes by injection of PGF2 alpha on days 5 to 10 of the estrous cycle (0 hr). Ewes received 0, 0.5, 1.0, 2.0 or 4.0 mg CGS per kg BW i.v. at -12, 0, 12 and 24 hr, and an ovulatory dose of hCG at 36 hr. Jugular (P < .001) and vena caval (P < .001) concentrations of E2 were decreased by CGS at all doses tested for 8 to 10 hr, but had returned to levels similar to control ewes by the time of the next injection. Concentrations of E2 around the time of the LH surge were similar in control and treated ewes. During the subsequent luteal phase, concentrations of progesterone (P4) were similar in control and treated ewes. Thus, transient decreases in E2 during the follicular phase were not deleterious to the subsequent luteal phase. In experiment 3, luteolysis was induced in 18 ewes by injection of PGF2 alpha on days 6 or 7 (0 hr) of the estrous cycle. Ewes received 0 or 1 mg CGS per kg BW i.v. every 8 hr from 0 to 40 hr. Ovulation was induced with hCG at 36 hr. CGS reduced jugular (P < .001) and vena caval (P < .001) concentrations of E2, prevented an endogenous surge of LH (P < .05) and increased (P < .001) concentrations of FSH. All ewes had ovulated a marked follicle by 72 hr, but onset of the luteal phase, as assessed by concentrations of P4, was delayed (P < .01) in ewes receiving CGS. Delayed luteal phases were not solely attributable to the presence of new CL or to luteinization of follicular cysts. When data were aligned according to the day ewes were observed in estrus, profiles of P4 did not differ with treatment.(ABSTRACT TRUNCATED AT 400 WORDS)

Two experiments (Spring and Fall) were conducted in ovariectomized ewes to determine changes in pituitary hormone secretion immediately after pituitary stalk-transection. Ewes underwent either pituitary stalk-transection (SS), sham-transection (SH) or administration of anesthesia only (AO). Stalk-transected, but not sham-operated or anesthetized ewes had polyuria and polydipsia for 7 to 14 days after surgery. Concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH) and prolactin were measured in peripheral blood samples collected every 10 minutes for a six-hour period. Results were comparable for each season. During the six hours following surgery or removal from anesthesia, concentrations of LH declined in all ewes, but more slowly in SS ewes. No differences in patterns or mean concentrations of FSH were observed. Immediately after surgery, concentrations of prolactin were elevated, then declined in SH and SS ewes. The decrease was greater in SH than SS ewes. Data are consonant with the view that hypothalamic inhibition as well as LHRH stimulation regulate gonadotropin release by the pituitary.

Three experiments were conducted to examine the relationship between systemic concentrations of luteinizing hormone (LH) and estradiol-17 beta (E2) after withdrawal of progesterone in cycling ewes. In Exp. 1, ewes were assigned randomly to one of three treatments: laparotomy (C), removal of the luteal ovary (ULO), or ULO plus anesthesia with sodium pentobarbital for 6 h beginning 4 h after surgery. Anesthesia was used in an attempt to block the expected increase in tonic secretion of LH. Patterns of LH and E2 in these three groups did not differ during the 24-h experimental period. In Exp. 2, a longer period of anesthesia was utilized. Forty-eight ewes were assigned at random to one of four treatments: C, ULO, lutectomy or an intrafollicular injection of prostaglandin F2 alpha (PGF2 alpha). One-half of the ewes in each group were anesthetized with sodium pentobarbital from initiation of treatment (0 h) until 10 h after surgery. Sodium pentobarbital did not suppress the increases in LH and E2 after progesterone withdrawal. The regression of concentrations of E2 on concentration of LH was not significant. In Exp. 3, ewes were infused with either saline or dopamine after receiving an im injection of PGF2 alpha. Tonic secretion of LH increased after 4 h in ewes infused with saline, but not in ewes infused with dopamine. Despite the suppression of LH, concentrations of E2 increased in dopamine-treated ewes as in control ewes. Therefore, the initial increase in E2 after a decline of progesterone in cycling ewes is independent of increases in LH.

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

In this study, we monitored episodic luteinizing hormone (LH) secretion throughout development in eight April-born ewe lambs to determine if a change in LH pulse patterns preceded first ovulation at puberty. LH pulses were measured in samples collected every 12 min for 6 h once in July, twice a month from 22 August to 2 October, and then weekly until puberty. Progesterone concentrations, measured in samples taken 3/wk, were used as an index of first ovulation, which occurred at 29.3 +/- 0.7 wk of age. LH pulse frequencies throughout most of this period ranged from 0 to 2 pulses/6 h, with no change over time. However, during the week prior to the first progesterone rise, there was a significant increase in pulse frequency to a level seen during the follicular phase in post-pubertal lambs. This increase in pulse frequency was evident in 7 of 8 lambs; pulses were not analyzed in the last lamb because samples were taken during the LH surge. In contrast, LH pulse amplitude did not increase prior to puberty. In fact, pulse amplitude declined linearly during the 3 wk before first ovulation and then increased during the follicular phase in post-pubertal animals. These results support the hypothesis that an increase in the frequency of episodic LH secretion is a key event leading to the onset of ovarian cycles in the lamb. Whether an increase in pulse amplitude is also necessary remains unclear. If so, it must occur just before the LH surge, since it was not detected in any samples taken before puberty in this study.

Two methods of induction of ovulation were examined in Manchega ewes that were in postpartum anestrus during the nonbreeding season. The experiment was of 2 X 2 X 3 factorial design. The methods compared were introduction of rams and this treatment plus an im injection of 50 micrograms luteinizing hormone releasing hormone (LHRH) at the time rams were introduced. Variations in response due to month of treatment (April, May or June) and interval from lambing to treatment (1, 2 or 3 mo) and their interactions with type of treatment were examined. Responses studied were proportions of ewes showing increases in plasma progesterone at (a) 10 d or (b) 17 or 24 d post-treatment, or lambing by 200 d post-treatment, and interval from treatment to lambing for ewes that did lamb by 200 d. The formation of a corpus luteum was determined by concentrations of progesterone in plasma; a positive response was considered to be a concentration greater than .5 ng/ml (baseline values averaged .1 ng/ml). Overall, there was no benefit of LHRH above the response to introduction of males only, in any trait examined. There was a significant interaction of treatment with month of treatment on the proportion of ewes forming corpora lutea by 17 or 24 d after initiation of treatment. This proportion was lower in June (38 vs 66% in April and 82% in May) for ewes receiving LHRH, but did not differ among months (61 to 68%) for ewes exposed to males only.(ABSTRACT TRUNCATED AT 250 WORDS)

Luteinizing hormone (LH) stimulates a cascade of ovarian hormonal events that culminate in ovulation. This study was designed to investigate, in sheep, sequential changes in prostaglandin (PG) E2, PGF2 alpha, 6-keto-PGF1 alpha, and cyclic adenosine monophosphate (cAMP) in the theca, granulosa and follicular fluid of large preovulatory follicles and small nonovulatory follicles in response to LH. On d 15 postestrus, preovulatory or nonovulatory follicles were injected intrafollicularly with saline or LH. Ewes were then ovariectomized at 0, 2, 4, or 8 h postinjection. Injected follicles were excised; theca, granulosa and fluid were separated, weighed and assayed for cAMP and PG. Contents of cAMP in the theca, granulosa and fluid of preovulatory follicles increased (P less than .01) 2 to 4 h after injection of LH. Increases (P less than .05) in contents of PGE2 and PGF2 alpha in the theca and fluid of preovulatory follicles were observed between 4 and 8 h after injection of LH. The time courses of LH-induced synthesis of PGE2 and PGF2 alpha in preovulatory follicles were parallel. Luteinizing hormone had no effect on PGE2, PGF2 alpha or cAMP in any compartment of small follicles. Contents of 6-keto-PGF1 alpha varied with time in both theca and granulosa of large and small, saline- and LH-injected follicles. Although specific increases in cAMP and PG followed an injection of LH only in large follicles, the parallel temporal relationship of PGE2 and PGF2 alpha did not explain the dichotomous functions ascribed to PGE2 and PGF2 alpha during the periovulatory period.

The effect of pregnancy on concentrations of prostaglandins E2, F2 alpha and 6-keto-prostaglandin F1 alpha (PGE2, PGF2 alpha and 6-keto-PGF1 alpha) in utero-ovarian venous plasma was examined in ewes on Days 10 through 14 after estrus, an interval which includes the critical period for maternal recognition of pregnancy. The utero-ovarian vein ipsilateral to a corpus luteum was catheterized on Day 9 or 10 in 6 pregnant and 8 nonpregnant ewes. Five blood samples were collected at 30-min intervals for 2 h beginning at 0500 and 1700 h daily. Sampling began at 0500 h on the day after catheterization. The mean and variance within each 2-h collection period were calculated for each ewe. The natural logarithm of the variance in each collection period (ln variance) was used as an estimate of the fluctuations in secretory activity by the endometrial-conceptus complex. Patterns of the mean concentrations of PGE2 were different between pregnant and nonpregnant ewes (P less than 0.01); PGE2 being higher in the pregnant ewes beginning on Day 13. There was a trend for the patterns of ln variance in PGE2 to differ (P less than 0.1) with pregnancy status over the entire period; ln variance was greater in pregnant ewes beginning on Day 13. The patterns of the mean concentrations and ln variances for PGF2 alpha and 6-keto-PGF1 alpha did not differ between pregnant and nonpregnant ewes. There were significant increases in both of these prostaglandins over time, independent of pregnancy status (P less than 0.01). The association of higher concentrations of PGE2 in utero-ovarian venous plasma with early pregnancy is consistent with the hypothesis that PGE2, originating from the uterus and/or conceptus, is one factor involved in maintenance of the corpus luteum of pregnancy.

Two experiments were conducted in peripuberal ewe lambs to determine (a) the influence of the first luteal structure [most frequently a transient (i.e., 1 to 4 d) structure] on subsequent ovarian activity and (b) a role for the uterus in its demise. In Exp. 1, 21 lambs were assigned randomly on the day of the first rise in progesterone in the plasma to (1) sham-operation, (2) removal of the nonluteal ovary, (3) removal of the luteal ovary and (4) removal of the luteal ovary plus progesterone replacement (5 mg given three times 12 h apart, initiated at surgery). No effect of treatment on subsequent ovarian activity was observed. In Exp. 2, four of 14 lambs were assigned randomly to be hysterectomized before their first rise in progesterone. The remaining 10 lambs, (five each) were sham-operated or hysterectomized on the day of the first rise in progesterone. All hysterectomized lambs (N = 9) exhibited a rise in progesterone and maintained elevated concentrations of progesterone, whereas sham-operated lambs initiated estrous cycles. Oviducts and uteri collected from lambs hysterectomized on the day of the first rise in progesterone in Exp. 2 were flushed for presence of oocytes and none were found. Similarly, no retained oocytes were found in histological preparations of first luteal structures obtained from eight lambs ovariectomized in Exp. 1. Concentrations of luteinizing hormone (LH) and prolactin were determined in daily samples collected from 19 lambs in Exp. 1. Luteinizing hormone increased and became more variable as lambs matured, whereas prolactin decreased with no detectable change in variability. It is concluded that the transient luteal structure is not required for sexual maturation and that its lifespan is uterine dependent.

The objectives of this study were to determine 1) the ability of charcoal-extracted bovine follicular fluid (bFF) to suppress endogenous follicle stimulating hormone (FSH) at various stages of the estrous cycle and 2) the effects of suppression of FSH on luteal function and lengths of the current and subsequent estrous cycles. Twenty-six mature ewes were assigned randomly to receive 5 ml of either bFF or saline, subcutaneously, at 8-h intervals on d 1 through 5 (bFF n = 6; saline n = 3), d 6 through 10 (bFF n = 6; saline n = 3) or d 11 through 15 (bFF n = 6; saline n = 2) of the estrous cycle (d 0 = estrus). Blood was collected daily beginning at estrus and continued until the third estrus (two estrous cycles) or 40 d; more frequent samples were collected 2 h prior to initiation of treatment (0600), hourly for the first 8 h of treatment, then every 4 h until 0800 on the first day after treatment, and finally at 1600 and 2400 on that day. Plasma concentrations of FSH were lower (P less than .001) in bFF-treated than in saline-treated ewes. Treatment with bFF reduced (P less than .05) plasma concentrations of progesterone during the current but not during the subsequent estrous cycle. Treatment with bFF did not affect plasma concentrations of estradiol-17 beta. Administration of bFF on d 11 through 15 of the estrous cycle lengthened the interval from the decline in progesterone to estrus and the inter-estrous interval by approximately 3 and 4 d, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)