Intrauterine infusion of BQ-610, an endothelin type A receptor antagonist, delays luteolysis in dairy heifers

Three separate in vivo experiments were conducted to evaluate the putative role of endothelin-1 (ET-1) during luteal regression in heifers. In Experiment 1, a single intraluteal injection of 500 μg BQ-610 [(N,N-hexamethylene) carbamoyl-Leu-d-Trp (CHO)-d-Trp], a highly specific endothelin A (ET_A) receptor antagonist, did not diminish the decline in plasma progesterone following a single exogenous injection of 25 mg prostaglandin F2 alpha (PGF₂) administered at midcycle of the estrous cycle. In Experiment 2, six intrauterine infusions of 500 μg BQ-610 given every 12 h on days 16–18 delayed spontaneous luteolysis, as evidenced by an extended elevation (P = 0.054) of plasma progesterone concentration. In Experiment 3, heifers were administered six intrauterine infusions of BQ-610 or saline on days 16–19, and peripheral blood samples were collected from day 11 to 16 (before infusion), hourly on days 16–19 (during infusion), and on days 20–25 (after infusion). BQ-610 treated heifers had markedly higher (P < 0.0001) levels of plasma progesterone compared with saline controls, and this effect was most notable during the infusion period (treatment by period interaction; P ≤ 0.05). Heifers infused with BQ-610 also had higher progesterone levels on day 21 (treatment by time interaction; P ≤ 0.05). Mean plasma concentrations of 13,14-dihydro-15-keto-PGF₂ (PGFM), the primary metabolite of PGF₂, were measured in the samples collected hourly and were not different (P ≥ 0.05) between treatments. These results indicate that the in vivo antagonism of the ET_A receptor can delay functional luteolysis, and supports the theory that ET-1 regulates luteal function in ruminants.     

Expression of mRNA for cell adhesion molecules in the bovine corpus luteum during the estrous cycle and PGF2alpha-induced luteolysis

Cell-to-cell interaction via cell contact-dependent pathway is essentially important for maintenance and regulation of corpus luteum (CL) integrity and its physiological actions. The objective of the present study was to evaluate the mRNA expression of the cell adhesion molecules (CAMs) that are constituent factors of gap junctions [connexin (Cx) 43] and adherence junctions (VE-, E-, N-cadherin) in two types of endothelial cells from the mid CL and in CL tissue during the estrous cycle and PGF(2alpha)-induced luteolysis in the cow. Specific mRNA expression for Cx43 and N-cadherin was detected in cytokeratin-positive (CK+) and cytokeratin-negative (CK-) luteal endothelial cells (EC) and fully luteinized granulosa cells (LGC). E-cadherin mRNA was expressed in CK+EC and LGC, but not in CK-EC. VE-cadherin mRNA was expressed in both CK+ and CK-EC. During the estrous cycle, Cx43 mRNA expression was significantly lower in the regressing CL. VE-cadherin expression also tended to increase in the mid CL and increased significantly in the regressing CL. E-cadherin mRNA expression was higher in the early and late CL than in the mid- and regressing CL. N-cadherin mRNA expression gradually increased from the early to late CL followed by a decrease in the regressing CL. During PGF(2alpha)-induced luteolysis, Cx43 mRNA expression appeared to increase, and VE-cadherin and E-cadherin mRNA significantly increased at 24 h. N-cadherin mRNA expression decreased 2 and 4 h after PGF(2alpha) administration. Collectively, expression of the mRNAs for CAMs was different in the two types of luteal endothelial cells and fully luteinized granulosa cells and changed independently in the CL during the estrous cycle and PGF(2alpha)-induced luteolysis in the cow. The results suggest that CAMs play physiological roles in cell-to-cell communication to regulate both gap and adherence junctions during CL development and regression in the cow.     

Effect of prostaglandin F2 alpha-induced luteolysis on in vivo and in vitro progesterone production by individual placentomes of cows

This study investigated placental progesterone production by bovine placentomes. Catheters were placed in the femoral artery (FA) and in the caruncular artery (CA), caruncular vein (CV) and lymphatic vessel of a prominent placentome of 13 cows at 200 d of gestation. Four of the 13 cows were given prostaglandin F2 alpha (PGF2 alpha) after surgery, and blood and lymph were collected for progesterone determination. After 24 h, progesterone was higher (P less than .01) in FA and CA plasma from control cows that FA and CA plasma from PGF2 alpha-treated cows (5.11 +/- .29 and 5.17 +/- .64 vs 1.41 +/- .08 and 1.15 +/- .08 ng/ml, respectively), but CV concentrations were similar (3.38 +/- .30 vs 2.56 +/- .24, respectively). There was a net uptake of progesterone by placentomes from control cows (P less than .01) but a net secretion in PGF2 alpha-treated cows (P less than .05). Lymph contained low progesterone concentrations regardless of treatment. Cows were slaughtered at 240 d of gestation. Placentomes were removed and perfused with pregnenolone through the maternal and fetal arteries. Fetal venous effluent contained more progesterone than maternal venous effluent (P less than .001) in both groups, and fetal venous effluent of placentomes from PGF2 alpha-treated cows contained more progesterone than that from control cows (P less than .05). Maternal and fetal components of other placentomes were cultured alone or in co-culture along with pregnenolone and (or) epostane. Fetal tissue produced more progesterone (P less than .001) than maternal tissue when each was cultured alone, but fetal tissue production declined when co-cultured with maternal tissue.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of an estrogen antagonist (tamoxifen) on cloprostenol-induced luteolysis in heifers

Experiments were conducted to determine the role of estrogens on endogenous PGF2 alpha secretion and luteolysis following injection of cloprostenol in heifers. In Exp. 1, eight luteal-phase heifers were used to evaluate tamoxifen (T) as an estrogen antagonist. Heifers received T (35 mg i.v.) or ethanol:saline vehicle (ES) every 4 h for 44 h. All received cloprostenol (500 micrograms i.m.) immediately after the start of T or ES, and received estradiol-17 beta (500 micrograms i.m.) 12 h later. Each ES heifer had a surge of luteinizing hormone (LH) within 48 h of estradiol injection, whereas T-treated heifers did not. Estrus was observed in three ES-treated heifers, but not in T-treated heifers. In Exp. 2, 10 heifers received T (35 mg i.v.) or ES every 4 h for 64 h beginning on d 15 postestrus. Cloprostenol (500 micrograms i.m.) was injected 16 h after the start of treatment. Concentrations of LH were similar (P greater than .05) in both groups. In ES heifers, concentrations of 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM) increased; in T-treated heifers, PGFM remained at pre-cloprostenol levels. Luteolysis was induced in all heifers. Progesterone (P4) decreased to less than or equal to 1 ng/ml at similar (P greater than .05) rates in ES-treated and T-treated heifers. Mean concentration of P4 288 h post-cloprostenol was greater (P less than .05) in ES-treated than in T-treated heifers. Three ES-treated heifers, but no T-treated heifers, were in standing estrus. We conclude that T effectively antagonizes estrogen in cattle.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of an oxytocin antagonist on prostaglandin F2 alpha secretion and the course of luteolysis in sows.

The role of oxytocin (OT) in the regulation of prostaglandin F2 alpha (PGF2 alpha) secretion during luteolysis in gilts was studied using a highly specific OT antagonist (CAP-581). In Experiment 1 gilts on Days 14 to 19 of the oestrous cycle in Latin square design were used, to determine the dose and time of application of OT and CAP. In Group I (n = 6) gilts were treated intravenously with saline or with 10, 20 and 30 IU of OT. Concentrations of the main PGF2 alpha metabolite i.e. 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM) were measured in blood samples as uterine response to the treatment. Twenty IU of OT was the most effective to stimulate PGFM release and this dose was used after CAP treatment in gilts of Groups II, III and IV. Gilts of Group II (n = 3) were injected into the uterine horns (UH) with saline (5 ml/horn) or CAP (2 mg, 3 mg and 4 mg; half dose/horn) and OT was injected (i.v.) 30 min thereafter. Any of the CAP doses given into the UH affected PGFM plasma concentrations stimulated by OT. In Group III (n = 4) gilts were infused (i.v.) for 30 min with CAP (9 mg, 14 mg and 18 mg/gilt) followed by 20 IU of OT. All doses of CAP effectively inhibited OT-stimulated PGF2 alpha release, therefore 9 mg was selected for the further studies. Gilts of Group IV (n = 4) received OT 4, 6 and 8 h after CAP to define how long CAP blocks the OT receptors. Concentrations of PGFM increased after any of this period of time. Thus, we concluded that 9 mg of CAP infused every 4 h will effectively block OT receptors. In Experiment 2, gilts (n = 4) received CAP as a 30-min infusion every 4 h on Days 12-20 of the oestrous cycle. Control gilts (n = 3) were infused with saline. CAP infusions diminished the height of PGFM peaks (P < 0.05). Frequency of the PGFM (P < 0.057) and OT (P < 0.082) peaks only tended to be lower in the CAP-treated gilts. Peripheral plasma concentrations of progesterone (P4) and oestradiol-17 beta (E2) and the time of luteolysis initiation as measured by the decrease of P4 concentration were the same in CAP- and saline-treated gilts. The macroscopic studies of the ovaries in gilts revealed lack of differences between groups. We conclude that OT is involved in the secretion of luteolytic PGF2 alpha peaks but its role is limited to controlling their height and frequency. Blocking of OT receptors did not prevent luteolysis in sows.     

Effect of PGF2 alpha administration on the release of endogenous PGF2 alpha and oxytocin in indomethacin-treated goats

Repeated intramuscular injection of 1 mg prostaglandin F2 alpha (PGF2 alpha) during the luteal phase of the oestrous cycle of the goat hastened luteolysis and resulted in rapid increases in jugular concentrations of 13,14-dihydro-15-keto PGF2 alpha (PGFM), the primary metabolite of PGF2 alpha, and of oxytocin; similar injections of PGF2 alpha in indomethacin-treated goats had a reduced effect on PGFM and oxytocin concentrations, and failed to induce luteolysis. The same injections of PGF2 alpha were without effect on PGFM and oxytocin concentrations in ovariectomised goats. The results suggest that exogenous PGF2 alpha, or endogenous PGF2 alpha released at luteolysis, may induce the release of ovarian oxytocin in the goat.     

Attenuation of PGF2alpha release in ewes infused with the oxytocin antagonist L-368,899

We have investigated the effects of systemic administration of the oxytocin antagonist (OTA) L-368,899 on luteolytic PGF(2alpha) release in ewes. In the first study, carried out in four ovariectomized ewes primed with progesterone to induce responsiveness to oxytocin, 3-h i.v. infusions of 3, 10 and 30 microg/kg/min OTA, carried out on days 12, 14, 16 and 18 in a Latin Square design, resulted in a significant attenuation of the oxytocin induced increase in PGFM concentration at all doses (OTA 139+/-8.3% of pre-oxytocin baseline; control 206.8+/-18.7%; P<0.005). In a further study, continuous infusion of cyclic ewes (n=6) with 10 microg/kg/min OTA from day 13 to day 17 of the cycle resulted in a reduction in both the frequency (OTA 1.0+/-0.4/ewe; control 2.2+/-0.2/ewe; P<0.05) and amplitude (OTA 31.8+/-11.0 pg/ml; control 68.8+/-10.4 pg/ml; P<0.05) of endogenous PGFM episodes compared to control ewes (n=5) measured during daily 8-h sampling windows on days 14-17. This reduction in PGFM concentrations was accompanied by a modest extension in the day of luteolysis (progesterone <0.5 ng/ml) to day 17.5+/-0.4 in the OTA treated group compared with day 16.4+/-0.5 in the control group (P=0.07). The results demonstrate that treatment with OTA caused a significant reduction in episodes of increased PGFM concentration during the period of luteolysis and may provide an approach by which to reduce early pregnancy failure.     

Carprofen in veterinary medicine. II. Inhibitory effect on the release of PGF2 alpha in the early postpartum cow

Carprofen, a non-steroidal anti-inflammatory drug (NSAID) known to inhibit prostaglandin synthesis, was given intravenously in five cows at a daily dose of 0.7 mg/kg for five days beginning on day 1 postpartum. Blood samples were collected at various times over a period of six days following the first injection. At this dose, carprofen reached highest plasma values of about 45 micrograms/ml after the fifth injection and was well tolerated by all the cows. During the whole experimental period, mean plasma levels of 15-keto-13, 14-dihydro-prostaglandin F2 alpha, the primary metabolite of PGF2 alpha, were significantly (p less than 0.05) lower in treated than in control animals (28-47% vs 64-101% of pretreatment concentrations). The suppressive effect of carprofen on PGF2 alpha-production occurred immediately after its application and was maximal 3-6 h post injection on the first and on the fifth experimental day (60-80% and 40-85%, respectively). We conclude from our results that carprofen in a single dose of 0.7 mg/kg b.w. effectively suppresses PGF2 alpha-release in the postpartum cow. Whether this effect is beneficial in the treatment of uterine inflammatory processes remains to be determined.     

Prostaglandin F2alpha (PGF2alpha) independent and dependent regulation of the bovine luteal endothelin system

We have examined the genes of the endothelin system that are targets for regulation by prostaglandin F2alpha (PGF2alpha). The effects of a luteolytic dose of PGF2alpha ) on the mRNA encoding endothelin converting enzyme-1 (ECE-1), pre-pro endothelin-1 (pp ET-1) and the ET receptors ETA, ETB, in bovine corpus luteum (CL) during the early (days 1 and 4), mid (day 10) or late (day 17) luteal phases were examined. The effect of the PGF(2alpha) treatment on ECE-1 protein, Big ET-1 and the biologically active mature ET-1 peptide were also examined. Most importantly, the direct ECE-1 activity was determined. Before day 10 of the cycle, in a PGF2alpha-independent manner, the amounts of mRNA encoding ET-1, ECE-1, ETA, and ETB were increased steadily from day 1. After day 10 of the cycle, expression of mRNA encoding pp ET-1 and ETA acquired responsiveness to exogenous PGF2alpha and both genes were up-regulated by the PGF2alpha treatment. This effect of PGF2alpha was also detected for the proteins corresponding to the mature ET-1. The enzymatic activity of ECE-1 remained unchanged throughout the lifespan of the CL in spite of the detected changes in mRNA and protein. The results suggest that the luteal endothelin system is regulated in a PGF2alpha-independent and -dependent manner. Importantly, an alteration in luteal ET-1 availability is most likely achieved by modulating the expression of mRNA encoding pp ET-1 and not by the amount or activity of ECE-1. This interpretation is supported by the observation that the activity of ECE-1 remained unchanged throughout the ovarian cycle. The combined effects of greater ET-1 availability and gene expression encoding the ETA receptor in the late luteal phase could render the CL, at this developmental stage, more sensitive or responsive to ET-1. If the luteal tissue is responsive to the available ET-1 during the early phase of the ovarian cycle, an additional role for ET-1 should be considered beyond mediating the luteolytic actions of PGF2alpha. Agents blocking the actions of ET-1 might be the best approach to interfere with the luteal ET system and test its physiological role(s) in vivo.     

Plasma concentrations of oxytocin,prostaglandin and ovarian steroids during spontaneous luteolysis in the cow

Fifteen, non-milking cyclic Holstein heifers and cows were used to study possible hormonal correlations during spontaneous luteal regression. Peripheral plasma samples were assayed for oxytocin, 13,14-dihydro-15-keto-prostaglandin F₂α (PGFM), estradiol-17β and progesterone. On day 9, when luteolysis does not normally occur, no significant elevation in oxytocin or PGFM values occurred in five cows sampled for 12 hr at 15 min intervals. A correlation did not exist (P>.05) between PGFM and oxytocin values nor between estradiol and oxytocin values at this time. Of 10 cows bled on day 18 or 19 for 12 hr at 15 min intervals or 34 hr at 1 hr intervals, five animals exhibited pulsatile elevations of PGFM and oxytocin. The elevations in plasma concentrations of these two hormones were coincident and significant correlation coefficients (P<.01) were calculated (range 0.62 to 0.85). No significant correlation was present in the remaining 5 cows. These results suggests that ovarian oxytocin is secreted concomitantly with uterine prostaglandin production at the time of spontaneous luteolysis in the cow.     

Effects of an endothelin receptor antagonist and nitroglycerin on digital vascular function in horses during the prodromal stages of carbohydrate overload-induced laminitis

OBJECTIVE: To evaluate changes in digital vascular function in horses with carbohydrate overload (CHO)-induced laminitis and determine the effects of an endothelin (ET) receptor antagonist and nitroglycerin on laminitis-associated vascular dysfunction. ANIMALS: 20 adult horses without abnormalities of the digit. PROCEDURES: Hemodynamic variables were recorded before (baseline) and hourly after all horses were administered a CHO ration via nasogastric tube. In 4 groups of 5 horses each, saline (0.9% NaCl) solution or ET receptor antagonist (10(5)M in digital blood) was administered into the digital arterial circulation according to 1 of 2 schedules. During anesthesia, blood flow; arterial, venous, and capillary pressures; and total, precapillary, and postcapillary resistances were measured in an isolated perfused digit of each horse. In all groups, nitroglycerin was infused (10(5)M in digital blood), and digital microvascular assessments were repeated. RESULTS: The CHO caused a significant decrease in right atrial pressure by 14 hours that was not affected by administration of saline solution or ET receptor antagonist. In isolated digits of anesthetized horses, CHO resulted in a significant decrease in digital blood flow associated with a significant increase in total and postcapillary resistances. Treatment with the ET receptor antagonist and nitroglycerin caused a significant decrease in total resistance. Postcapillary resistance was significantly decreased following treatment with the ET receptor antagonist but was not altered by treatment with nitroglycerin. CONCLUSIONS AND CLINICAL RELEVANCE: Treatment with an ET receptor antagonist and nitroglycerin resulted in significant improvement in vascular resistance in isolated perfused digits of anesthetized horses with CHO-induced laminitis.     

育成和经产母牛肌肉注射PGF2a同期发情对比试验

[目的] 探讨应用氯前列烯醇诱导母牛同期发情的方法。 [方法] 选择育成母牛和经产母牛,一次性肌肉注射2支(0.2mg/支),通过发情数量、时间、同期率、受配率等情况对比同期发情率的差异。 [结果] 结果表明,在牛体格、生理状况和饲养管理条件大体一致的情况下,育成牛发情比经产牛早3-5h;在发情持续时间上,育成牛比经产牛早结束2-3h,症状显著,而相对持续时间较短,受配率育成牛多于经产牛。 [结论] 用一次PG法处理繁殖母牛可获得较高的同期发情率,育成牛同期发情效果稍优于经产母牛,体况因素也是母牛发情的重要影响因素之一。     

Epidural injection of ketamine for caudal analgesia in the cow

The objective of the study was to determine the analgesic and sedative effects of epidural ketamine in the cow. Eight healthy cows weighing 350–450 kg were used. One of 3 doses of ketamine (0.5, 1 and 2.0 mg/kg) or a saline control were injected into the epidural space at the first intercoccygeal interspace in random sequence at one-week intervals. Ketamine was diluted in saline (0.9%) before the experiment, and the volume adjusted according to animal size. Analgesia was tested by applying a standard stimulus (needle insertion into the skin and deep muscle) and scored using a 3-point scale. A second voltage-based stimulus was also applied and the responses scored. Another scale was used for scoring the degree of sedation. The response and the degree of sedation were assessed before drug administration and at 2, 5, 10, 15 min, and every 15 min until 120 min after ketamine or saline administration. Ketamine produced dose-related analgesia of the tail, anus, perineum and vulva but not of the hindlimb area. The effect was dose-dependent in terms of intensity and duration. None of the doses produced analgesia when 70 or 80 volts were applied. Minimal side effects were observed. Epidural ketamine produces caudal analgesia in the cow. Further studies are required to determine whether this is sufficient for surgery.     

Local mechanisms for luteolysis in the cow: Novel roles of vasoactive substances in the luteolytic cascade within the corpus luteum

The corpus luteum (CL) in the estrous cycle in the cow is a dynamic organ which has a lifespan of approximately 17–18 days. As the CL matures, the steroidogenic cells establish contact with many capillary vessels and the CL is composed of a large number of vascular endothelial cells that can account for up to 50% of the bovine CL. Furthermore, luteal cells and endothelial cells secrete several vasoactive substances such as prostaglandin F_2α (PGF_2α), endothelin‐1 and angiotensin II. These vasoactive substances also function in regulating progesterone secretion in an autocrine/paracrine manner in the CL. The blood vessels and endothelial cells in the CL therefore have an essential role in the luteal function in the cow. Endometrial PGF_2α, the primary luteolysin in the cow, stimulates luteal vasoactive substances during luteolysis. Moreover, luteal vasoactive substances may have key roles in the regulation of luteolysis to induce vasodilatation, vasoconstriction and angiolysis. This review describes the current concept for possible roles of vasoactive substances in the luteolytic cascade within the bovine CL.     

Effects of a histamine type 2 receptor antagonist, BMY-26539-01, on equine gastric acid secretion.

A dose-response study was undertaken of the effects of a newly developed histamine type 2 receptor antagonist, BMY-26539-01, on gastric acid secretion in 4 fasted horses. Doses of 0.1 mg/kg, 0.3 mg/kg, 0.5 mg/kg, or placebo were administered in a randomly assigned treatment sequence. Hydrogen ion concentration and pH were variable during baseline measurements in all 4 animals; however, following BMY-26539-01 administration, mean pH increased and hydrogen ion concentration decreased in a dose-related pattern. At the 0.3 mg/kg and 0.5 mg/kg dose levels, pH remained elevated for > 4 h and > 8 h, respectively. No adverse effects were observed. A significant level of 0.01 was used for all statistical methods.     

Effects of TAK-044, a nonselective endothelin receptor antagonist, on the spontaneous and indomethacin- or methylene blue-induced constriction of the ductus arteriosus in rats

We studied the effects of TAK-044, a nonselective endothelin (ET) receptor antagonist, on the indomethacin- or methylene blue-induced constriction of the ductus arteriosus (DA) in rats and compared them with the effects on spontaneous DA constriction. Injection of TAK-044 into 21-day-old fetuses in utero was performed through the uterine wall of laparotomized mother rats under light ether anesthesia. The fetuses were autopsied 3 hr after treatment with TAK-044 (10 mg/kg) in utero and simultaneous administration to the laparotomized mother rats of indomethacin (3 mg/kg, p.o.) or methylene blue (100 mg/kg, i.p.). In the second experiment, pregnant rats were decapitated on day 21 of gestation to obtain newborn rats by cesarean delivery. Newborn rats which were given TAK-044 (2, 10 mg/kg) immediately after or 1 hr before cesarean delivery were autopsied at various times after birth. In both experiments, pups were rapidly frozen in an acetone-dry ice mixture at autopsy to evaluate the DA constriction by the whole-body freezing and shaving method. TAK-044 injection into the fetus 3 hr before autopsy completely inhibited the DA constriction induced by maternal treatment with indomethacin or methylene blue. TAK-044 caused dose-dependent inhibition of the spontaneous closure of the DA after birth. The inhibitory effect was more pronounced in pups which were given TAK-044 in utero 1 hr before birth; however, the inhibitory effect was incomplete in newborn pups. These results, together with the previous finding that BQ-123, an ETA-specific receptor antagonist, inhibits the ductal constriction induced by oxygen in vitro [Coceani et al., 1992], indicate that the ETA receptor plays a significant role in the indomethacin- or methylene blue-induced DA constriction as well as in the spontaneous DA constriction after birth, and also indicate that the inhibition of ETA receptor by TAK-044 was more easily achieved in fetuses than in neonates.     

Effect of the administration of PGF2 alpha synchronously with insemination on the pregnancy rate in mares in an insemination program

Investigations in different species including the horse have demonstrated that prostaglandin F2 alpha (PGF2 alpha) is involved in initiating uterine contractions occurring during mating and artificial insemination (A.I.). Uterine contractions play an important role with respect to the sperm transport within the female genital tract. The objective of the present investigation was to evaluate whether the administration of PGF2 alpha (Dinoprost) synchronously to A.I. could have a positive effect on the pregnancy rate in mares. A field study including 346 warmblood-mares (age two to 20 years) belonging to a private studfarm was conducted during the breeding season 1996. The mares were assigned to two groups, group A: mares with spontaneous ovulation, group B: mares in which the ovulation was induced by a GnRH-analog-implant (Deslorelin). PGF2 alpha (Dinoprost) was administered either intramusculary (i.m., 5.0 mg) or intrauterine (i.ut., 0.5 mg diluted in 1.9 ml isotonic NaCl-solution and added to the semen dosis). The study was carried out in a double-blind fashion using isotonic NaCl-solution as a placebo. The mares of each group were randomly assigned to one of the two treatments (i.m. vs. i.ut.). The following first cycle pregnancy rates (day 18) were obtained in mares treated and inseminated once per oestrus: group A1 (PGF2 alpha, i.m.): 54.5% (n = 33); group A2 (placebo, i.m.): 69.7% (n = 33); group A3 (PGF2 alpha, i.ut.): 65.4% (n = 26); group A4 (placebo, i.ut.): 69.8% (n = 32); group B1 (PGF2 alpha, i.m.): 56.5% (n = 46); group B2 (placebo, i.m.): 29.6% (n = 27); group B3 (PGF2 alpha, i.ut.): 66.7% (n = 45); group B4 (placebo, i.ut.): 60.0% (n = 30). The pregnancy rates did not differ between the different groups with the exception of group B2 (p < 0.05). In mares treated repeatedly during the oestrus period (group A, n = 88; group B, n = 23), the pregnancy rates did not differ significantly between treatment and control groups. From the results obtained it is concluded that the PGF2 alpha-application did not show an effect on the pregnancy rate. Further factors influencing the results to a small degree were the stallions, semen age and quality and frequency of insemination per oestrus.