The effect of estrogen, progesterone and prostaglandin F2 alpha on uterine contractions in seasonally anovulatory mares

Uterine contractions were studied in two experiments utilizing ultrasonography and seasonally anovulatory mares. A one-minute ultrasound scan was done to produce longitudinal real-time images of the uterine body and an overall uterine contractile activity score (0 = no or minimal activity to 4 = maximal activity) was assigned to each scan. In experiment 1, a two-hour uterine activity trial (one score every 10 minutes) was done in mares given a single injection of prostaglandin F2 alpha (PGF2 alpha group; n = 4) and in control mares (n = 4). There was no difference between the two groups over the two-hour trial (mean activity score averaged over the two-hour trial: PGF2 alpha group, 0.2; control group, 0.1). In experiment 2, 16 mares were randomly assigned to one of four groups: 1) controls (corn oil vehicle), 2) 1 mg estradiol 17 beta on days 0 to 9 and 100 mg progesterone on days 10 to 20 (E2--greater than P4 group), 3) 100 mg progesterone on days 0 to 20 (P4 group), and 4) 100 mg progesterone on days 0 to 9 and 1 mg estradiol 17 beta + 100 mg progesterone on days 10 to 20 (P4--greater than E2 + P4 group). Uterine activity was assessed for each mare daily. The day by group interaction was significant. Scores for the E2--greater than P4 group were greater on days 4 to 11 (P less than .05) than for the other three groups. From day 14 to 21, scores did not differ among the three steroid-treated groups (except on day 15), but the scores averaged over each steroid-treated group were greater for each day (P less than .1 or .05) than for the controls (except on day 17).(ABSTRACT TRUNCATED AT 250 WORDS)     

Relationship between peripartal plasma oxytocin and prostaglandin F(2alpha) metabolite and placental expulsion time in heavy draft mares

The aim of this study was to clarify the relationship between circulating oxytocin (OT) and PGF(2alpha) metabolite (PGFM) in mares at the third stage of labor and placental expulsion time in order to investigate a cause of retained placenta of which the incidence increase in a heavy draft mare. Blood was sampled every 5 min from foaling to expulsion of the placenta in 18 heavy draft mares to evaluate circulating OT and PGFM. The relationships between OT and PGFM concentration and recorded placental expulsion times were investigated. The results were as follows (1) The highest level of OT concentration was observed close to foaling in 15 mares. (2) The OT concentrations close to foaling were variable with a large difference from the lowest concentration, 22.1 pg/ml, to the highest concentration, 209.3 pg/ml. (3) The highest level of PGFM was observed close to foaling in 17 mares. (4) During the 60 min following foaling, the OT concentrations of the mares (n=11) that had a shorter placental expulsion time (i.e., <1 h), were significantly higher than those of the mares (n=7) that had a longer placental expulsion time (i.e., >1 h; P<0.05). Collectively, the OT concentration immediately after foaling is negatively related to the placental expulsion time. Deficiency of OT secretion at foaling have should be considered as one of the causes of retained placenta in heavy draft mares.     

Acquisition of luteolytic capacity involves differential regulation by prostaglandin F2alpha of genes involved in progesterone biosynthesis in the porcine corpus luteum

Luteolytic capacity is defined as the ability of corpora lutea (CL) to undergo luteolysis after prostaglandin (PG) F2alpha treatment. The mechanisms causing acquisition of luteolytic capacity are not yet identified but CL without luteolytic capacity have PGF2alpha receptors and respond to PGF2alpha with some changes in gene expression. Inhibition of progesterone biosynthesis is a key feature of luteolysis and therefore we postulated that genes involved in progesterone biosynthesis would be regulated by PGF2alpha differently in CL with or without luteolytic capacity. Gilts on day 9 after estrus (lack luteolytic capacity) or day 17 of pseudopregnancy (with luteolytic capacity) were treated with saline or a PGF2alpha analog (cloprostenol) and CL were collected 0.5 (Experiment I) or 10 h (Experiment II) later. In Experiment III, large luteal cells from CL on day 9 or 17 were cultured for 1, 12 and 24h with or without PGF2alpha. PGF2alpha decreased LDL receptor mRNA (27%), steroidogenic acute regulatory protein (StAR) mRNA (41%), StAR protein (75%), LH receptor mRNA (55%), and LH receptor protein (45%) at 10 h after treatment in day 17 but not day 9 CL. PGF2alpha increased DAX-1 mRNA at 0.5 h (43%) and 10 h (46%) after PGF2alpha in day 17 but not day 9 CL but decreased 3betaHSD mRNA ( approximately 20% at 10 h) in both days 9 and 17 CL. In vitro, PGF2alpha decreased StAR mRNA at 12 h only in day 17 luteal cells; however, continuous treatment with PGF2alpha for 24 h decreased StAR mRNA in both days 9 and 17 luteal cells. Thus, luteolytic capacity involves a critical change in responsiveness of DAX-1, StAR, and LH receptor to PGF2alpha that results in inhibition of luteal progesterone biosynthesis.     

Changes of plasma concentrations of steroid hormones, prostaglandin F2 alpha-metabolite and pregnant mare serum gonadotropin during pregnancy in thoroughbred mares.

Plasma concentrations of estrogens, gestagens, cortisol (F), 13, 14-dihydro, 15-keto PGF2 alpha (PGFM) and pregnant mare serum gonadotropin (PMSG) in 10 Thoroughbred mares were measured for a 11-month pregnancy period. Estrone (E1) and estradiol-17 beta (E2) levels gradually increased as the pregnancy advanced, showing a peak around Month 8 and decreased thereafter. Progesterone (P) levels increased on Months 3 and 11, and 17 alpha-OH-progesterone (17 alpha-OHP) levels peaked on Month 3, whereas 20 alpha-OH-progesterone (20 alpha-OHP) levels increased sharply after Month 6. PGFM indicated peaks on Months 2 and 11. F and PMSG levels peaked on Months 2 and 3. From factor analysis, Month, E1, E2 and 20 alpha-OHP were discriminated as Factor 1, increasing with the progress of pregnancy, PMSG, 17 alpha-OHP and P as Factor 2, showing a relation with the secondary corpus luteum, and PGFM and F as Factor 3, affecting PGFM change on Month 2 by F. P also related to both Factors 1 and 3, showing an inverse relation against PGFM. In conclusion E1, E2 and 20 alpha-OHP contained in Factor 1 were suggested to be important especially as parameters of placental function after Month 6.     

Correlations among body temperature,plasma progesterone,cortisol and prostaglandin F2alpha of the periparturient bitch

The results of this study suggest that, besides the irrelevant role of body temperature measurement to predict the impending parturition in the bitch, progesterone and 15-ketodihydroprostaglandin F2alpha plasma level records could be more suitable to detect the approaching whelping in this species. More interesting was the statistically significant substantial increase in body temperature beginning 12 h after the onset of parturition. Therefore, if any significant increase in body temperature is recorded at the end of pregnancy without the beginning of the expulsion of fetuses, it could indicate problems at parturition. In this study, cortisol levels increased significantly at the time of delivery and remained high 12 h after the beginning of parturition, decreasing within 36 h after the onset of whelping. 15-ketodihydro-prostaglandin F2alpha levels increased significantly 24 h before parturition and again at the onset of whelping. Progesterone levels decreased significantly, starting 24 h before the onset of whelping and remained low after delivery.     

The hormone profile in the blood plasma of heifers after cycle synchronization with prostaglandin F2alpha

Highly sensitive radio-immuno assay (RIA) was used for 21 days to study the curves of several steroid hormones, progesterone (P), oestradiol 17 beta (E2), and testosterone (T), as well as of luteinizing hormone (LH) in peripheral blood of six heifers to which oestrus had been induced by prostaglandin F2 alpha. The validity of the authors' RIA for P, E2, and T determination in blood plasma was positively verified by two reliability criteria, correctness and accuracy, as well as by comparative determinations, using reference methods. Only four of the six heifers returned to oestrus, within 21 days from first oestrus induction. A typical cycle-related curve of the P concentration in peripheral blood with peak values between the 13th and 17th days of cycle (15-26 nmol/l) was recorded from four animals. The peak values of pre-ovulatory E2 and LH were between 33.0 and 53.2 pmol/l or between 19.5 and 52.5 micrograms/l. Some of the T rises in peripheral blood were in parallel to E2 concentrations. All hormone curves are presented in detail and are discussed in relation to clinico-physiological findings.     

Regulation of endometrial prostaglandin F(2alpha) synthesis during luteolysis and early pregnancy in cattle

Luteal regression is caused by a pulsatile release of prostaglandin (PG) F(2alpha) from the uterus in the late luteal phase in most mammals including cattle. Although it has been proposed in ruminants that pulsatile PGF(2alpha) secretion is generated by a positive feedback loop between luteal and/or hypophyseal oxytocin and uterine PGF(2alpha), the bovine endometrium may possess other mechanisms for initiation of luteolytic PGF(2alpha) secretion. It has been recently demonstrated that tumor necrosis factor-alpha (TNF-alpha) stimulates PGF(2alpha) output from bovine endometrial tissue not only during the follicular phase but also during the late luteal phase, suggesting that TNF-alpha is a factor in the initiation of luteolysis in cattle. Furthermore, our recent study has shown that IFN-tau suppresses the action of TNF-alpha on PGF(2alpha) synthesis by the bovine endometrium in vitro, suggesting that IFN-tau plays a luteoprotective role by inhibiting TNF-alpha-induced PGF(2alpha) production in early pregnancy. On the other hand, factors other than oxytocin or TNF-alpha have also been suggested to be involved in the regulation of PGF(2alpha) synthesis by bovine endometrium. The purpose of this review is to summarize our current understanding of the endocrine mechanisms that regulate the timing and pattern of uterine PGF(2alpha) secretion during the estrous cycle and early pregnancy.     

Oxytocin stimulates secretion of prostaglandin F(2alpha) from endometrial cells of swine in the presence of progesterone

Oxytocin (OT) stimulates endometrial secretion of prostaglandin (PG) F(2 alpha) during corpus luteum regression in swine but there is differential responsiveness to OT among endometrial cell types. To determine if progesterone influenced responsiveness of luminal epithelial, glandular epithelial, and stromal cells to 100 nM OT during luteolysis in swine, cells were isolated from endometrium of 15 gilts by differential enzymatic digestion and sieve filtration on day 16 postestrus and cultured continuously in the presence of 0, 10 or 100 nM progesterone. For phospholipase C (PLC) activity and PGF(2 alpha) secretion, stromal cells were most responsive to OT (P<0.01) in the absence of progesterone, whereas luminal epithelial cells were unresponsive and glandular epithelial cells displayed an intermediate response to OT (P<0.09). Progesterone enhanced PLC activity linearly in glandular epithelial cells (P<0.05) and influenced it quadratically in stromal cells (P=0.05). The effect of OT and progesterone on PLC activity in luminal epithelial cells was not significant, and progesterone did not increase PLC activity in response to OT in any cell type. Culture in the presence of progesterone, enhanced PGF(2 alpha) secretion in response to OT in luminal epithelial cells (P<0.05) but not in glandular epithelial or stromal cells. Progesterone also increased overall PGF(2 alpha) release from glandular epithelial (P<0.05) and stromal cells (P<0.06) across both levels of OT treatment. These results indicate that progesterone enhanced PGF(2 alpha) secretion from luminal epithelial cells in response to OT and increased basal PGF(2 alpha) release from glandular epithelial and stromal cells.     

Effects of GnRH or progesterone treatment on day 5 post-AI on plasma progesterone, luteal blood flow and leucocyte counts during the luteal phase in dairy cows

This study was designed to test the effects of progesterone or GnRH treatment on day 5 post-AI on fertility and luteal function in dairy cows and heifers. Five days after AI, 32 animals were randomly assigned to a control, intravaginal progesterone for 14 days progesterone releasing intravaginal device (PRID) or GnRH treatment group. On days 5, 7, 12, 14, 17 and 19 post-AI, each animal underwent colour Doppler ultrasonography of the corpus luteum and blood samples were collected for cell counts and plasma progesterone determination. Through general linear model repeated measures analysis of variance, significant effects were observed of treatment, parity, inseminating bull, reduced vascularization of the CL and pregnancy on plasma progesterone concentrations, whereas mean plasma progesterone and time luteal phase day, and treatment and plasma progesterone concentration on day 5 post-AI were found to, respectively, affect neutrophil and lymphocyte counts throughout the luteal phase. Moreover, two binary logistic regression analyses were performed. Based on the odds ratio, the likelihood of pregnancy by days 26-32 post-AI was 23.4 times higher in animals with high mean progesterone levels throughout the study period, compared with animals with low mean progesterone. The likelihood of reduced CL vascularization was 14 times higher in animals treated with PRID, compared with control and GnRH-treated animals. In conclusion, our results indicate that treatment on day 5 post-AI with PRID reduced subsequent CL vascularization, whereas GnRH treatment increased plasma progesterone concentrations on day 12 post-AI, although an effect was identified of the inseminating bull on plasma progesterone levels. Pregnant animals showed higher mean plasma progesterone concentrations than in nonpregnant ones and heifers higher than in lactating cows, whereas blood cell counts differed depending on the treatment and on the mean plasma progesterone concentration on day 5 post-AI.     

Plasma levels of cortisol, progesterone, oestradiol-17 beta and prostaglandin F2 alpha metabolite after ACTH (Synacthen Depot) administration in ovariectomized gilts

In order to elucidate the effect of stress on reproductive hormones, the present study was designed to investigate the effect of adrenocorticotropic hormone (ACTH) on the plasma levels of cortisol, progesterone, oestradiol-17 beta and prostaglandin F2 alpha metabolite in ovariectomized gilts. Ovariectomy and cannulation of the jugular vein were performed within 1 week of oestrous detection, under general anaesthesia. Approximately 1 week after surgery, two gilts were each administered ACTH (Synacthen Depot) intravenously, at a dose of 0.01 mg/kg body weight, and one gilt was given saline solution (5 ml). The reverse was performed on the following day. The administration of ACTH was followed by a concomitant elevation of cortisol, progesterone and prostaglandin F2 alpha metabolite but not of oestradiol-17 beta. Peak cortisol, progesterone and prostaglandin F2 alpha metabolite levels were reached at 80 +/- 10.0, 80 +/- 10.0 and 46.6 +/- 13.3 min after ACTH administration and the durations of the peaks were 181.8 +/- 19.8, 308.1 +/- 49.7 and 181.8 +/- 7.9 min, respectively. The total area under the curve for cortisol, progesterone and prostaglandin F2 alpha metabolite was significantly higher in the ACTH than in the control group. The present results indicate that during stress, cortisol, progesterone and prostaglandin F2 alpha levels are elevated while the level of oestradiol-17 beta is less affected. It can be concluded that the administration of ACTH to ovariectomized gilts, results in the elevation of cortisol, progesterone and prostaglandin F2 alpha metabolite but not of oestradiol-17 beta.     

Investigation into the use of prostaglandin F2 alpha (PGF2 alpha) and oxytocin for the induction of farrowing

This experiment was conducted in order to compare the effects of injecting PGF2 alpha alone, PGF2 alpha with oxytocin and placebo on the induction of farrowing in swine and to compare the relative effects of 3 different dosages of oxytocin (10, 20 and 30 iu per animal) when combined with PGF2 alpha (10 mg). The findings revealed that animals treated with 30 iu oxytocin farrowed within 10.6 h which was similar to those receiving PGF2 alpha only (9.4 h), but shorter than control animals (53.6 h). Animals receiving 20 and 10 iu of oxytocin farrowed within 1.4 and 1.7 h, respectively. Difficult farrowings requiring manual assistance occurred in 30%, 30%, 50% and 10% of sows given 30 iu, 20 iu and 10 iu of oxytocin and in the control group, respectively. Thirteen of 73 sows treated with PGF2 alpha farrowed within 12.6 +/- 5.3 h. Stillbirths were highest (10.2%) in the control animals whilst in the others it was under 7%. Oxytocin at dosages of 20 and 10 iu, seemed most promising in terms of synchronising farrowing following PGF2 alpha treatment in swine. However, farrowing complications were more common in these groups.     

Resynchrony of previously anoestrous cows and treatment of cows not detected in oestrus that had a palpable corpus luteum with prostaglandin F(2 alpha)

AIMS: To determine if resynchrony, using a progesterone (P4) / oestradiol benzoate (ODB) regime, of previously treated, anovulatory anoestrous (AA) cows would increase the probability of oestrus, conception and pregnancy compared to no resynchrony. Additionally, the effect of prostaglandin F2alpha (PG) treatment of cows not detected in oestrus, but in which a corpus luteum (CL) was palpable transrectally (NDO/CL+), was compared with no treatment. METHODS: Cows not detected in oestrus by 1 week before the start of the seasonal breeding programme were categorised as AA or NDO/CL+, based on absence or presence of a palpable CL, respectively. All AA cows were treated with an intravaginal device containing 1.36 g P4 (CIDR-B) for a period of 6 days, and with 1 mg ODB by intramuscular (I/M) injection 1 day after the device was removed (Day 0). Half the AA cows that were detected in oestrus between Days 0 and 3 were randomly assigned to be resynchronised by reinsertion of a clean used CIDR-B device on Day 15, for a period of 6 days, followed by an I/M injection of 0.5 mg ODB, 1 day after the device was removed (resynchrony, RS), while the other half were not resynchronised (no-RS). NDO/CL+ cows were alternately assigned to be either treated with 25 mg of the PG, dinoprost tromethamine, on Day 0 or left as untreated controls (Con). RESULTS: Resynchrony increased the percentage of cows detected in oestrus between Days 14 and 28 (212/282 = 75.2% vs 155/285 = 54.4%, for RS vs no-RS, respectively; p<0.001), but had no effect on conception rate to a service within the first 3 days of the mating period (146/397 = 36.8% vs 148/399= 37.1%, for RS vs no-RS cows, respectively; p>0.9), or to a service between Days 14 and 28 (84/159 = 52.8% vs 114/217 = 52.5%, for RS vs no-RS cows, respectively; p>0.9). Resynchrony increased the Day 28 pregnancy rate (264/432 = 61.1% vs 237/435 = 54.5%, for RS vs no-RS cows, respectively; p=0.03), but had no effect on the Day 56 or final pregnancy rates (p>0.1).Prostaglandin treatment of NDO/CL+ cows did not affect the percentage of cows detected in oestrus by Day 7 (43/106 = 40.6% vs 51/101 = 50.5%, for Con vs PG, respectively; p=0.15), or Day 28 (92/106 = 86.8% vs 91/101 = 90.1%, for Con vs PG, respectively; p>0.4). Treatment did not affect the Day 28 pregnancy rate (55/103 = 53.4% vs 54/98 = 55.1%, for Con vs PG, respectively; p>0.9), the Day 56 pregnancy rate (81/103 = 78.6% vs 74/98 = 75.5%, for Con vs PG, respectively; p>0.6), or the final pregnancy rate (98/103 = 95.1% vs 89/97 = 91.8%, for Con vs PG, respectively; p>0.3). CONCLUSIONS: Resynchrony of AA cows treated using the present protocol increased the proportion of non-pregnant cows detected in oestrus between Days 14 and 28 and increased the Day 28 pregnancy rate. This resynchrony protocol may be useful for increasing the proportion of the herd pregnant in the first month of the breeding programme, especially in herds that have a history of a low proportion of non-pregnant cows detected in oestrus between Days 14 and 28. There was no benefit in treating NDO/CL+ cows with PG compared to no treatment. KEYWORDS: Dairy cows, anoestrus, treatment, resynchrony, prostaglandin, progesterone, oestradiol benzoate.