Unilateral luteotropic effect of uterine venous effluent of a gravid uterine horn in sheep.

The involvement of the main uterine vein in the unilateral maintenance of CL was studied in bilaterally ovulating unilaterally pregnant ewes. Ewes were mated at estrus (Day 0) and bilaterally ovulating ewes were randomized into three groups at surgery on Day 5. In all ewes, the uterine horns were separated through the intercornual area and one was ligated and transected near the internal bifurcation to produce a nongravid horn. One group served as controls (five ewes). In the other two groups the main uterine vein on one side was surgically anastomosed (end to side) to the corresponding vein of the opposite side (gravid side to nongravid side in one group--three ewes, and nongravid side to gravid side in the other--three ewes). Necropsies were done on Day 20. Mean CL weight was less, (P less than .01) on the nongravid side in control ewes than on the gravid side in control ewes or for either side in the other two groups. There were no significant differences among mean weights of CL on the gravid side in control ewes and either side in the other two groups. The CL regressed when the ipsilateral uterine vein contained blood from only the nongravid horn whereas the CL was maintained when the ipsilateral uterine vein contained venous blood from a gravid horn, whether or not it also contained blood from a nongravid horn. Results indicate that the uterine venous effluent from a gravid uterine horn in sheep has a luteotropic effect on the ipsilateral CL.     

Artificial insemination of ewes with frozen-thawed semen at a synchronised oestrus. 2. Effect of dose of spermatozoa and site of intrauterine insemination on fertility

Three experiments were conducted to examine the effect of dose of inseminate, number of uterine horns inseminated and site of insemination on subsequent fertility of Merino ewes after synchronisation of oestrus, with progestagen-impregnated sponges (inserted for 12 days) and an injection of PMSG, and intrauterine insemination with frozen-thawed semen.The percentages of ewes lambing after insemination with 0.5, 5, 25 and 50 × 10⁶ spermatozoa were 29.3, 26.8, 56.3 and 62.1% respectively. A similar trend was observed in a second test resulting in 23.5, 38.8 and 53.1% ewes lambing after insemination with 5, 10 and 20 × 10⁶ spermatozoa respectively.The percentage of ewes lambing was higher for ewes inseminated in two uterine horns than one horn (76.8 vs. 44.9, P < 0.001). When semen was deposited in the tip, middle and bottom of the uterine horn, the percentages of ewes lambing and lambs born per ewe inseminated were 43.6 and 52.7, 52.8 and 84.9, and 41.2 and 64.7% respectively. Although site of insemination did not affect the percentage of ewes lambing, the percentage of lambs born per ewe inseminated was higher after insemination in the middle of the uterine horn than at the other sites (P < 0.001).     

Effects of postparturient uterine lavage on uterine involution in the mare

Eighteen postparturient mares were used to evaluate effects of uterine lavage on uterine involution. Mares were randomly assigned to one of three treatment groups: Group 1 (seven mares), no lavage; Group 2 (five mares), lavage on Day 3 post partum; and Group 3 (six mares), lavage on Days 3, 4, and 5 post partum. Five liters sterile physiologic saline, prewarmed to 42 degrees C, were used for each lavage. Transrectal ultrasound examination of the reproductive tract was performed on Day 11 post partum to detect the presence of free fluid in the uterine lumen, to estimate the cross-sectional diameter of the uterine horns and body, and to determine if ovulation had occurred. Endometrial biopsies were also taken on Day 11 post partum to evaluate endometrial histologic characteristics. Lavage had no effect (P>0.05) on diameter of the uterine body or previously gravid uterine horn, presence of fluid in the uterine lumen, or number of mares which had ovulated by Day 11 post partum. Histologic characteristics of the endometrium (height of luminal epithelium, gland depth, relative gland vclume, and inflammatory-cell score) were not affected by treatment (P>0.05). Postpartum uterine lavage did not significantly affect uterine involution by the parameters measured in normal-foaling mares at Day 11 post partum.     

Effects of ecbolic agents on measurements of uterine involution in the mare

Thirteen postparturient mares were used to investigate the effects of ecbolic agents on the rate of uterine involution. Mares were randomly assigned to one of three treatment groups: Group S = intravenous injection of 2 ml saline twice daily for 10 days post partum (n=4); Group O = intravenous injection of 20 units oxytocin twice daily for 10 days post partum (n=4); and Group P = intramuscular injection of 500 mcg fluprostenol twice daily for 10 days post partum (n=5). Ovulation was determined by daily transrectal ultrasonographic examination of the ovaries. On Days 6, 11 and 16 post partum, transrectal ultrasonography was used to measure cross-sectional diameters of the uterine body, uterine horns and fluid within the uterine lumen. Uteri were swabbed for aerobic bacteriologic culture on Days 11 and 16 post partum. Uterine biopsies were obtained from the base of the previously gravid uterine horn on Days 11 and 16 post partum for subjective assessment of endometritis and morphometric analysis of endometrial histoarchitecture. Mean values for all measurements of uterine involution did not differ among groups (P>0.05). For all mares, the diameter of luminal fluid was not correlated to diameter of the uterine body or uterine horns, or to morphometric measurements of endometrial histoarchitecture of the previously gravid uterine horn (P>0.05). Likewise, accumulation of fluid within the uterine lumen was not associated with endometritis or recovery of potential bacterial pathogens (P>0.05). Mean diameter of the previously gravid uterine horn was negatively correlated with morphometric measures of endometrial histoarchitecture of the previously gravid uterine horn (P<0.01).     

Effects of season and lactation on luteinizing hormone secretion in postpartum ewes

Effects of season, postpartum interval and short-term weaning were investigated on luteinizing hormone (LH) secretion in ewes. Blood samples were collected at 10-min intervals for 4 h (basal period). Then gonadotropin-releasing hormone (GnRH) was administered and 10 more blood samples were collected over an additional 4 h period. The effects of day post partum (5, 20 or 40) and short-term weaning (weaned Day 37, tested Day 40 post partum) on basal and GnRH-induced LH secretion were tested. Mean basal concentrations of LH for ewes on Day 5, 20 or 40 post partum ranged from 1.6 to 4.6 ng/ml and did not differ. Mean concentrations of LH during the post-GnRH sampling interval were greater (P<0.01) for ewes bled on Day 20 or 40 post partum (12.3 and 11.8 ng/ml, respectively) than for ewes bled on Day 5 or for unbred control ewes (6.7 and 5.8 ng/ml, respectively). Weaning on Day 37 depressed GnRH-induced LH secretion on Day 40 post partum (8.18 ng/ml; P<0.05). Seasonal changes in LH secretion on Day 20 or 40 post partum in January, March or June lambing ewes were also tested. There was no difference in basal or GnRH-induced LH secretion between Day 20 or 40 post partum among groups in January or March.. In June, ewes had lower (P<0.01) basal and GnRH-induced LH secretion on Day 20 post partum than ewes did on Day 40 post partum. Across month of the year, on Day 20 post partum, ewes lambing in March released more LH in response to GnRH than ewes lambing in January (P=0.07) or June (P<0.05). Response to GnRH on Day 20 post partum was similar for ewes lambing in January or June (P>0.1). Ewes lambing in January released less (P<0.01) LH on Day 40 post partum than ewes lambing in March or June; however, no difference was detected between the latter two groups (P>0.1). Thus, seasonal modifications of the releasable pool of LH may mask or modify the effect of the postpartum interval upon this endocrine response.     

Resumption of ovarian follicular activity and uterine involution in the postpartum llama

Resumption of ovarian follicle activity and uterine involution was studied in the post partum llama. Thirty-nine adult multiparous llamas were monitored by ultrasonography and analysis of urinary estrone sulfate for 30 d post partum at the La Raya Research Station in Peru. Uterine involution was measured in terms of reduction of length and diameter of both uterine horns. Correlation analysis was used to relate follicle size and concentration of estrone sulfate. Analysis of variance was used to determine rate of uterine involution relative to days post partum. The left ovary was palpated and scanned by Day 3 post partum in contrast to Day 1 post partum for the right ovary. Ovulatory size follicles, 7 mm, were present by Day 7.4 post partum (range 4 to 14 d). Follicle growth was detected as early as Day 4 post partum with follicle size being less during the first follicle wave (7.4 mm) compared to the second and third waves (9 to 10 mm). Concentrations of urinary estrone sulfate were positively related (P<0.05) to follicular size, but to a lesser degree during the first follicle wave (19.4 ng/mg Cr), than to the second wave (25.4 ng/mg Cr). Uterine involution, as measured by diameter, was different between the left (gravid) and right (nongravid) uterine horn (P<0.05) for the 17 d post partum, and was also different from that of control females for the 21 d post partum. Uterine involution was complete in 63% of females by Day 21 post partum.     

Comparing early embryo mortality in dairy cows during hot and cool seasons of the year

The objectives of this study were to identify the level and stage of embryonic mortality that occur in dairy cows during hot and cool seasons of the year. Experimental dairy cows, of varying ages, were artificially inseminated with frozen-thawed semen from proven Holstein sires. Females on each dairy unit were then randomly allocated to one of three experimental groups after partitioning by day of artificial insemination, days post partum, parity, and current milk production level. In Group I and Group II, nonsurgical embryo collection was performed on each cow using Dulbecco's phosphate-buffered saline as the flushing medium. Embryos from cows in Group I were collected on Days 6 or 7 post insemination during the hot (n=93) and cool (n=64) seasons. Embryos from cows in Group II were collected on Days 13 or 14 post insemination during the hot (n=97) and cool (n=63) seasons. In Group III, contemporary control cows were also inseminated during the hot (n=106) and cool (n=106) seasons, and fetal heart beat was evaluated via ultrasound between Days 25 and 35 following insemination. Embryo viability decreased (P<0.05) from 59% at Day 7 to 27% at Day 14 in the hot season, but was not decreased during the cool season (52 vs. 60%). Pregnancy rate at Days 25 to 35 was 21% in the hot season, which was less (P<0.05) than the 36% in the cool season. The percentage of unfertilized ova collected in both the hot and cool seasons suggests that fertilization failure was not affected by season of breeding. In summary, embryonic loss after Day 7 of pregnancy appears to be a problem in this hot, dry climate.     

Biodegradable estradiol microspheres do not affect uterine involution or characteristics of postpartum estrus in mares

Quarterhorse mares were used to investigate effects of estradiol-17beta on uterine involution, duration of estrus, interval to ovulation, and fertility achieved by breeding on the first postpartum estrus. On the day of foaling, mares were injected with biodegradable poly (DL-lactide) microspheres containing either 100 mg estradiol-17beta (25 mares) or no drug (27 mares). The treatment period was considered to last for 12 to 15 d. Estrus was determined by teasing mares (n=16) with a stallion. Ovulation was detected by transrectal ultrasonographic examination of ovaries (n=48). On Days 6, 11 and 16 post partum, transrectal ultrasonography was used to measure cross-sectional diameters of the uterine body, uterine horns, and fluid within the uterine lumen (n=28). Uteri were swabbed for bacteriologic culture, and uterine biopsies were obtained from the previously gravid uterine horn on Days 11 and 16 post partum, for assessment of endometritis and morphometric analysis of endometrial histioarchitecture (n=19). Twenty-two mares were bred on foal-heat, and pregnancy was determined by transrectal ultrasonography on 14 to 16 and 30 to 35 d after breeding. With only one exception (diameter of previously gravid uterine horn on Day 11), mean values for all measures of uterine involution did not differ between treatment groups (P > 0.05). No differences were detected between treatment group means for length of estrus or interval to ovulation (P > 0.05). No differences were detected between treatment group liklihoods for recovery of potential bacterial pathogens, presence of endometritis, or presence of intrauterine fluid at 11 or 16 d post partum (P > 0.05). Pregnancy rate of mares treated with estradiol (5 11 ; 45%) was not different from that of control mares (9 11 ; 82%; P > 0.05). Estradiol treatment did not hasten uterine involution, increase duration of estrus, delay ovulation, or increase fertility in these postpartum mares.     

Successful pregnancies after transfer of embryos recovered from ewes induced to ovulate 24-29 days post partum

After lambing in late November, oestrus and ovulation were induced by using a CIDR device and PMSG in early weaned (N = 13) or lactating (N = 14) Border Leicester x Scottish Blackface ewes between 23 and 29 days after parturition. Ewes were intrauterine inseminated under laparoscopic visualization 54-55 h after CIDR-device withdrawal and eggs recovered on Day 3 of the cycle. Ovum recovery and fertilization rates were higher in lactating than in early weaned ewes, with fertilization being achieved as early as 24 days post partum in both groups. Of the 7 early weaned and 11 lactating ewes yielding eggs, fertilization occurred in 4 and 7 ewes respectively. A total of 20 embryos were transferred to the normal uterine environment of 15 recipient ewes in which the interval from parturition was greater than 150 days. Pregnancies were successfully established in 9 recipient ewes, resulting in the birth of 10 viable lambs. Prolactin concentrations were significantly higher (P less than 0.001) in lactating than in early weaned ewes throughout the study. Nevertheless, normal luteal function (as assessed by daily progesterone concentrations) was exhibited by 12 of 14 lactating and 8 of 13 early weaned ewes. Two post-partum donors in which the corpora lutea completely failed to secrete progesterone yielded fertilized eggs which developed to term when transferred to a normal uterine environment. The results show that sheep oocytes can be fertilized using laparoscopic intrauterine insemination as early as 24 days after parturition and that the resulting embryos are viable when recovered on Day 3 after oestrus and transferred to a normal uterine environment.     

Production of prostaglandin f2alpha and its metabolite by endometrium and yolk sac placenta in late gestation in the tammar wallaby, Macropus Eugenii

In this study, we investigated production of prostaglandin (PG) F2alpha and its metabolite, PGFM, by uterine tissues from tammar wallabies in late pregnancy. Endometrial explants were prepared from gravid and nongravid uteri of tammars between Day 18 of gestation (primitive streak) and Day 26.5 (term) and were incubated in Ham's F-10 medium supplemented with glutamine and antibiotics for 20 h. PGF2alpha and PGFM in the medium were assayed by specific, validated RIAs. Control tissues (leg muscle) did not produce detectable amounts of either PG. Both gravid and nongravid endometria secreted PGF2alpha, and production increased significantly in both gravid and nongravid uteri towards term. PGFM was produced in small amounts by both gravid and nongravid uteri, and the rate of production did not increase. Neither oxytocin nor dexamethasone stimulated PG production in vitro in any tissue at any stage. Thus, the surge in peripheral plasma PGFM levels seen at parturition may arise from increased uterine PG production, but further study is needed to define what triggers this release.     

How the FSH/LH ratio and dose numbers in the p-FSH administration treatment regimen, and insemination schedule affect superovulatory response in ewes

We wished to evaluate the effects of FSH/LH ratio and number of doses of p-FSH during a superovulatory treatment on ovulation rate and embryo production (Experiment I). In Experiment II, we studied the efficacy of fertilization after various insemination schedules in superovulated donors. In Experiment I estrus was synchronized in 40 ewes (FGA, for 9 days plus PGF2alpha on Day 7) and the ewes were randomly assigned to four treatment groups as follows (n = 10 ewes each): Group A: four p-FSH doses with the FSH/LH ratio held constant (1.6); Group B: four p-FSH doses with the FSH/LH ratio decreasing (FSH/LH 1.6-1.0-0.6-0.3); Group C: eight p-FSH doses with the FSH/LH ratio held constant (1.6); Group D: eight p-FSH doses and FSH/LH ratio decreasing (1.6-1.6, 1.0-1.0, 0.6-0.6, 0.3-0.3). p-FSH administrations were performed twice daily 12 h apart. The ewes were mated at the onset of estrus and again after 12 and 24 h; then, one ram per four ewes was maintained with the ewes for two additional days. Ovarian response and embryo production were assessed on Day 7 after estrus. Experiment II. Three groups (n = 10 each) of superovulated ewes were inseminated as follows: Group M: mated at onset of estrus; Group AI: artificial insemination 30 h after onset of estrus; M + AI) mating at onset of estrus and intrauterine AI performed 30 h from estrus with fresh semen. Results of Experiment I showed that treatment (D) improved (P < 0.05) ovulatory response in comparison to Groups (C) and (A). The fertilization rate was lower (P < 0.01) in Group D) than Group (A). Also the proportion of transferable embryos was lower in Group (D) in comparison to all the other treatments (P < 0.01). Group A gave the best production of embryos (7.3/ewe; 89.0% transferable). In Experiment II, combined mating plus AI improved fertilization rate (80.3%) compared to both mating (P < 0.01) and AI (P < 0.02) alone.     

The effect of postpartum uterine lavage on foal heat pregnancy rate

Mares (n = 37) were treated on Days 2 and 4 post partum with a uterine lavage of 10 l of warm, sterile NaCl (0.9%) solution. Endometrial cytology and culture were performed on Day 7. Mares were bred on the first postpartum estrus by artificial insemination. Pregnancy rates were determined by ultrasound examination at Day 16 post ovulation. No differences were noted in degree of uterine inflammation or presence of uterine bacteria at Day 7 post partum between treated (n = 18) and control (n = 19) mares. Pregnancy rates at the first postpartum estrus for treated mares (55.5%) was not statistically different from that of control mares (68.4%). No advantage was noted in the use of intrauterine lavage with 10 l of warm sterile NaCl (0.9%) at Days 2 and 4 post partum as a means of improving foal heat pregnancy rate.     

Growth and microvascular development of the uterus during early pregnancy in ewes.

Growth and microvascular development of the uterus were evaluated for ewes on Days 12, 18, 24, and 30 after mating (3-4 ewes/day; Day 0 = day of mating) in two experiments. In experiment 1, fresh weight and dry weight of gravid uterine horns were increased on Days 24 and 30 after mating, whereas those of nongravid uterine horns were elevated only on Day 30. The increased fresh and dry weights of gravid uterine horns on Day 24 were associated with uterine hyperplasia (increased DNA content). Increased fresh and dry weights of gravid uterine horns on Day 30, however, were associated with hypertrophy (increased RNA:DNA and protein:DNA ratios) of uterine tissues. In experiment 2, vascularity of endometrial tissues was elevated on Days 24 and 30 after mating. In addition, dramatic changes in uterine architecture (increased lumenal diameter and decreased endometrial thickness) and in uterine microvascular development (increased abundance of large microvessels and development of a subepithelial capillary plexus) were observed by Day 24 after mating. Characterization of the patterns of uterine growth and microvascular development will enable us to further define the role of previously reported uterine and conceptus-derived growth and angiogenic factors during early pregnancy.     

Estrus, ovulation and conception following timed insemination in Romney ewes treated with progestagen and gonadotropins.

The estrus — ovulation time relationships was examined in Romney ewes treated with progestogen (intravaginal sponge) and gonadotropins (PMSG + HCG or PMSG alone) prior to (January) and during (April) the breeding season. The conception rate of ewes inseminated at predetermined times after treatment was also investigated.Ewes exhibited estrus sooner after sponge removal in April than in January (34.9 v 38.9 hrs, P < 0.001). The interval from sponge removal to ovulation was also shorter in April than in January (56.3 – 62.1 hrs, P < 0.01). There were no significant differences between treatments or season on the mean interval from estrus to ovulation. Types of gonadotropin treatment had no effect on the estrus — ovulation time relationships. There were no significant effects of season, hormone treatment or time of insemination on lambing rate.     

Effect of breeding season on fertility of sheep following estrus synchronization and fixed-time artificial insemination under field conditions in semi-arid tropical region

Accelerated lambing system is heavily reliant on reproductive technologies meant to enable off the season breeding in sheep. Therefore, the present study was programmed to assess the effect of breeding season (BS) on fertility of sheep following estrus synchronization (ES) and fixed time artificial insemination (FTAI). A total of 248 and 365 ewes were synchronized and inseminated during the BS (Febuary–March and July–September) and non-breeding season (NBS) respectively, during 2012–14. Synchronization of estrus was done by AVIKESIL-S, intra-vaginal progesterone sponges kept in situ in vagina for 12 days. 200 IU eCG was administered intramuscularly on 12th day after sponge withdrawal. FTAI was performed twice in ewes exhibiting signs of estrus at 48 and 56h after sponge removal, using liquid chilled semen of Patanwadi/Malpura rams containing 100 million sperm. Breeding season had no significant (p<0.05) effect on estrus synchronization. The estrous responses during the BS and NBS were 84.68% and 83.29% respectively. The lambing percentage during BS was 66.67%, which is significantly (p<0.05) higher than the lambing percentage of NBS (57.57%). Although, the lambing percentage in NBS maneuvered ewes were lower than the BS ewes but still the technology can be used to offset the effect of anestrus and to augment production in sheep.     

Side of gestation in dairy heifers affects subsequent sperm transport and pregnancy rates after deep insemination into one uterine horn

Effects of side of previous gestation on sperm transport and pregnancy rates after deep cornual insemination were evaluated in 1686 Friesian cows in their first lactational period. Only single ovulating animals were used. At insemination, semen was deposited deep into the uterine horn ipsilateral or contralateral to the preovulatory follicle. A total of 876 cows (52%) ovulated in the ovary ipsilateral to the postgravid horn, and 810 cows ovulated in the contralateral ovary. Semen was deposited into the previously nongravid uterine horn of 832 cows, and into the gravid horn of 854 cows. The pregnancy rate was higher (P < 0.00001) for semen deposition into the previously nongravid horn (46.6%) than for semen deposition into the gravid horn (35.7%). For inseminations ipsilateral to the side of impending ovulation, pregnancy rates were higher (P = 0.0004) when ovulations occurred on the opposite side to the postgravid horn than on the same side. Pregnancy rates were higher (P = 0.002) for contralateral inseminations when ovulations occurred on the same side to the postgravid horn than on the opposite side; they were higher (P = 0.0001) for total ipsilateral than for total contralateral inseminations. There was no difference between ipsilateral and contralateral inseminations (P = 0.64) when ovulation occurred ipsilateral to the postgravid horn, but pregnancy rates were higher (P < 0.00001) when ipsilateral insemination was carried out into the nonpostgravid horn. Results indicate that the side of gestation in dairy heifers affects subsequent pregnancy rates after deep insemination into one uterine horn, possibly by affecting sperm transport.     

Lambing rates and litter sizes following intrauterine or cervical insemination of frozen/thawed semen with or without oxytocin administration

Intrauterine insemination by laparoscopy is required to achieve acceptable lambing rates in ewes when using frozen semen but the procedure has evoked welfare concerns. Oxytocin has been used to dilate the cervix as a means of accessing the uterus during conventional cervical insemination, but its effect on fertility is not well documented. Three hundred crossbred ewes were synchronised in estrus and randomly allocated to one of three insemination procedures using frozen/thawed semen containing 400 x 10(6)/ml progressively motile sperm: single cervical (0.2 ml), multiple cervical (4 x 0.05 ml) or laparoscopic (0.05 ml per uterine horn). The effects of each insemination procedure on lambing rate (percentage of treated ewes lambing) and litter size (lambs per ewe lambing) were tested with and without oxytocin (10 IU given i.m.) prior to fixed-time insemination. Oxytocin did not permit complete cervical penetration in any ewes and neither lambing rate nor litter size was influenced by the number of inseminations. Lambing percentages were 69 and 42 (P < 0.01) for the laparoscopic and cervical insemination methods, respectively, and oxytocin reduced these to 58 (NS) and 10 (P < 0.001) percent, respectively. Corresponding litter sizes for ewes not receiving oxytocin were 1.91 and 1.51 and for those receiving oxytocin, 1.83 and 1.41 (laparoscopic versus cervical, P < 0.02). Thus, in the absence of complete cervical penetration at insemination, 10 IU oxytocin decreased the number of ewes lambing but had no effect on their litter size.     

Unilateral uteroovarian relationship in pregnant cattle and role of uterine vein

The hypothesis of a unilateral uteroovarian relationship between location of embryos and maintenance of corpora lutea (CL) was tested in superovulated cattle. Ovulations were induced in both ovaries and a uterine horn was isolated subsequent to insemination to produce unilateral pregnancy. The mean weight of CL on day 24 was greater (P<.05) on the gravid side (2580 mg) than on the nongravid side (1200 mg; n=5), supporting the hypothesis. Involvement of the main uterine vein in the unilateral pathway between a gravid horn and the adjacent ovary was demonstrated in an experiment which utilized separation of uterine horns, anastomosis of uterine veins between sides, and insertion of embryos into one side by embryo transfer. Mean weight of CL on day 24 was less (P<.05) when the gravid horn was contralateral to the CL (950 mg; n=3), than when the gravid horn was ipsilateral to the CL (4760 mg) or when the gravid horn was contralateral to the CL and the uterine vein from the contralateral side was anastomosed to the vein on the ipsilateral side (3580 mg; n=3).     

Combination of the ram effect with PGF2α estrous synchronization treatments in ewes during the breeding season

The role of the ram effect on the reproductive performance of ewes that have initiated estrous cycles following lambing in combination with synchronization of estrus using PGF(2α) was examined. A total of 1264 Corriedale × Merino ewes in the breeding season (March-April) were allocated to one of three treatments. The control group (PG2) of ewes (n=415) were in permanent direct contact with vasectomized rams throughout the experiment from 60 d prior to the administration of the first luteolytic dose of PGF(2α) which was followed by a second dose 13 d later (Day 0 of the experiment). Ewes assigned to the other two treatments remained isolated from rams until Day 0. In the second treatment, ewes (PG2RE; n=445) were administered PGF(2α) in the same manner and were joined with vasectomized rams at Day 0. Ewes allocated to the third treatment (PGRE; n=404) did not receive the second dose of PGF(2α) but were introduced to vasectomized rams on Day 0. Sexual receptivity, as indicated by tail-head marking, was recorded until d 11. More PG2RE ewes (407/445; 92%) were observed in estrus by Day 11 than occurred for PG2 ewes (353/415; 85%; P=0.003). The accumulated frequency of PG2RE ewes in estrus was greater than for PG2 ewes for each period from Day 3 (P<0.001) to Day 11 (P<0.01). The onset of estrus was earlier in PG2RE ewes (2.98±0.07 d) than for PG2 ewes (3.31±0.07 d; P<0.0001). In contrast, the total frequency of PGRE ewes observed in estrus by Day 11 (356/404; 88%) was similar to that observed for PG2 ewes. However, the trajectory of the accumulated frequency of the incidence of estrus was less for the PGRE ewes initially, particularly during the period of Days 3-6 of observation (P<0.0001). Consequently, onset of estrus was earlier in PG2 ewes (3.31±0.07 d) than for PGRE ewes (5.30±0.11 d; P<0.0001). It was concluded that the introduction of vasectomized rams simultaneously with the second administration of PGF(2α) advanced the onset of estrus and increased the number of ewes that responded. The introduction of rams 13 d after a single dose of PGF(2α) did not substitute for the second administration of PGF(2α).     

Progesterone and placentation increase secreted phosphoprotein one (SPP1 or osteopontin) in uterine glands and stroma for histotrophic and hematotrophic support of ovine pregnancy

Secreted phosphoprotein one (SPP1, osteopontin) may regulate conceptus implantation and placentation. We investigated effects of progesterone (P(4)) and the conceptus on expression and localization of SPP1 in the ovine uterus. Steady-state levels of SPP1 mRNA in the endometrium of unilaterally pregnant ewes did not differ significantly between nongravid and gravid horns within their respective days of pregnancy; however, levels did increase as pregnancy progressed. SPP1 mRNA was detectable in the glandular epithelium (GE) of both nongravid and gravid horns via in situ hybridization. SPP1 protein was localized to the apical surface of the luminal epithelium of both nongravid and gravid uterine horns. Gravid horns exhibited extensive stromal SPP1 on Days 40 through 120, whereas SPP1 was markedly lower in the stroma of nongravid uterine horns through Day 80 of pregnancy. By Day 120, stromal expression of SPP1 between nongravid and gravid horns was similar. Long-term P(4) treatment of ovariectomized ewes induced SPP1 in the uterine stroma and GE. A bioactive 45-kDa SPP1 fragment was purified from uterine secretions and promoted ovine trophectoderm cell attachment in vitro. Interestingly, increased stromal cell expression of SPP1 was positively associated with vascularization as assessed by von Willebrand factor staining. Finally, ovine uterine artery endothelial cells produced SPP1 during outgrowth into three-dimensional collagen matrices in an in vitro model system that recapitulates angiogenesis. Collectively, P(4) induces and the conceptus further stimulates SPP1 in uterine GE and stroma, where SPP1 likely influences histotrophic and hematotrophic support of conceptus development.