Oxytocin modulates the pulsatile secretion of prostaglandin F2alpha in initiated luteolysis in cattle

Subluteolytic doses of prostaglandin F2alpha analogue (oestrophan) given i.m. and oxytocin (OT) antagonist (CAP) and noradrenaline (NA) infused into the abdominal aorta were used to test the importance of luteal OT in pulsatile secretion of prostaglandin F2alpha (PGF) during luteolysis in heifers (n = 17). In experiment 1, heifers were pre-infused for 30 minutes with saline on either day 17 of the oestrous cycle (group 1; n = 4) or on day 18 of the oestrous cycle (group 2; n = 3), and with CAP (8 mg per animal) on day 17 of the oestrous cycle (group 3; n = 4). Next, heifers were injected with oestrophan (30 microg per animal). Injection of oestrophan in Group 3 increased OT concentrations (P < 0.001) to values similar to those observed during spontaneous luteolysis (50 to 70 pg ml(-1)). PGFM concentrations in this group also increased (P < 0.001), but were lower (P < 0.05) than the values in groups 1 and 2, CAP given prior to oestrophan decreased both PGFM elevation (P < 0.06) and its area under the curve (P < 0.01), compared to the saline pretreated heifers. In experiment 2 NA (4 mg) was infused twice for 30 minutes at five hour intervals to release OT on day 17 of the oestrous cycle (n = 6). However, during hormone analysis it appeared that three of six heifers had elevated PGFM concentrations (group 1) and three others did not (group 2). NA caused the correlated increase of progesterone and OT secretion (r = 0.68; P < 0.05) in both groups but it only influenced PGF secretion in group 1 only (P < 0.05). We postulate that OT can amplify and modulate the course of induced luteolysis as a regulator of the amplitude of pulsatile PGF secretion. PGF analogue stimulates secretion of endogenous PGF from the uterus in cattle and this may be an important component of the luteolytic response to exogenous PGF.     

Oxytocin-induced changes in plasma 13,14 dihydro-15-keto prostaglandin F2 alpha concentrations on days 10, 20 and 30 postpartum in the bovine

Eighteen suckled Brahman cows were allotted randomly to treatments arranged in a three-period crossover design according to calving date and prior treatment such that each cow received 30, 150 and 300 IU oxytocin (OT) i.v. on d 10, 20 or 30 postpartum. Blood was collected via an indwelling jugular catheter every 15 min for 195 min. Samples collected before OT administration were used to determine basal plasma 13,14-dihydro-15-keto prostaglandin F2 alpha (PGFM) concentration. Day, time and the day X dose interaction affected PGFM (P less than .0001). All doses of OT elevated PGFM on all days postpartum (P less than .0001). Basal PGFM was greater (P less than .0001) on d 10 (252.2 +/- 51.2 pg/ml) than on d 20 (78.2 +/- 14.8 pg/ml) or on d 30 (64.8 +/- 7.4 pg/ml). The rise in PGFM in response to OT was greatest on d 10 and decreased (P less than .001) with increasing days postpartum. On d 10, 150 IU of OT caused a greater (P less than .0007) rise in PGFM than either 30 or 300 IU. On d 20, the 300-IU dose raised PGFM more (P less than .005) than either 30 or 150 IU, whereas on d 30 no differences among doses were detected. Cows had higher basal PGFM and a greater response to OT on d 10 postpartum than on d 20 or 30; cows were more responsive on d 20 than on d 30. All doses of OT elevated PGFM at all three times postpartum; however, differences between doses were not detected.(ABSTRACT TRUNCATED AT 250 WORDS)     

Luteolysis and associated interrelationships among circulating PGF2α, progesterone, LH, and estradiol in mares

The changing concentrations and temporal relationships among a PGF2α metabolite (PGFM), progesterone (P₄), LH, and estradiol-17β (E₂) before, during, and after luteolysis were studied in 10 mares. Blood samples were collected every hour for ≥4 d beginning on day 12 after ovulation. The luteolytic period extended from a decrease in P₄ at a common transitional hour (Hour 0) at the end of preluteolysis and beginning of luteolysis to a defined ending when P₄ reached 1 ng/mL. The length of luteolysis was 22.9 ± 0.9 h, contrasting with 2 d in published P₄ profiles from sampling every 6 to 24 h. In mares with complete data for Hours −40 to −2 (n = 6), PGFM concentrations remained below assay sensitivity (n = 2) or two or three small pulses (peak, 29 ± 4 pg/mL) occurred. During luteolysis, the pulses became more prominent (peak, 193 ± 36 pg/mL). Rhythmicity of PGFM pulses was not detected by a pulsatility program during preluteolysis but was detected in seven of nine mares during luteolysis and postluteolysis combined. The nadir-to-nadir interval for LH pulses and the peak-to-peak interval between adjacent pulses were longer (P < 0.05) during preluteolysis than during luteolysis (nadir to nadir, 5.2 ± 0.3 h vs 3.6 ± 0.4 h; peak to peak, 9.4 ± 1.0 h vs 4.7 ± 0.5 h). Unlike reported findings in cattle, concentrations of P₄ decreased linearly within the hours of each PGFM pulse during luteolysis, and a positive effect of an LH pulse on P₄ and E₂ concentration was not detected. The reported balancing of P₄ concentrations between a negative effect of PGF2α and a positive effect of LH in heifers was not detected in mares.     

Nocturnal melatonin and prolactin plasma concentrations in sheep selected for fertility in autumn lambing'.

Ewes selected for fertility in autumn lambing were used to evaluate correlated responses in nocturnal hormone levels. Four jugular blood samples were obtained during nighttime in August from each of 113 selected and 69 control ewes. Melatonin levels were lower for selected ewes (143 +/- 14 pg/mL) than for control ewes (184 +/- 13 pg/mL), and melatonin levels decreased with increases in estimated breeding values (EBV) for fertility (-2.23 +/- 0.79 pg mL(-1) x %(-1)). Prolactin levels were higher for selected ewes (90 +/- 7 ng/mL) than for control ewes (52 +/- 7 ng/mL), but significant line x ewe age interaction was also observed, with smaller differences in prolactin levels in 2-yr-old and older ewes (74 +/- 7 vs. 56 +/- 9 ng/mL for select and control ewes, respectively; P < 0.20 before and P = 0.05 after logarithmic transformation). Prolactin levels increased with both fertility EBV (1.23 +/- 0.53 ng mL(-1) x %(-1)) and maternal birth weight EBV (9.0 +/- 4.0 ng x ml(-1) kg(-1)). Heritability estimates were 0.43 (P < 0.02) for melatonin levels and 0.11 (P > 0.25) for prolactin levels. Thus, we conclude that selection for fertility in autumn lambing has affected patterns of melatonin and prolactin secretion during the dark phase.     

Effect of progesterone supplementation on pregnancy and embryo survival in ewes

Two hundred eighteen ewes were used in experiments 1) to develop a progesterone supplementation regimen capable of sustaining serum concentrations of progesterone at about 2.0 ng/ml for a period of 50 d (Exp. 1) and 2) to determine the effects of progesterone supplementation (d 6 to 50 after mating) on pregnancy and embryo survival rates in mated ewes (Exp. 2). In ovariectomized ewes in Exp. 1, s.c. administration of four cylindrical (9.5 x 60 mm) silastic implants, containing 20% (1.1 g) progesterone by weight, sustained mean serum concentrations of progesterone of 1.9 +/- .07 ng/ml compared with 1.03 +/- .05 ng/ml in ewes bearing two implants. In Exp. 2 each ewe (n = 159) was mated to two fertile rams at a spontaneous estrus (d 0) during mid-breeding season. Mean ovulation rate, determined on a subgroup of 46 ewes, was 1.45 +/- .05. On d 6, ewes were assigned randomly to control (two implants containing no progesterone) or progesterone-treated (four implants similar to those used in Exp. 1) groups. From d 7 to 50 after mating, progesterone concentrations in serum were greater (P less than .001) in progesterone-treated (four implants similar to those used in Exp. 1) groups. From d 7 to 50 after mating, progesterone concentrations in serum were greater (P less than .001) in progesterone-treated (3.50 +/- .06) than in control (2.65 +/- .05) ewes. Pregnancy rates (86% and 83%) and calculated embryo survival rates (77% and 78%) were similar (P greater than .05) for the control and progesterone-treated groups, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Reproductive characteristics of endophyte-infected or novel tall fescue fed ewes

Reduced pregnancy rates often occur in ruminants grazing endophyte-infected (EI) tall fescue. The objectives were to characterize basal and oxytocin-induced PGF_2α concentrations in serum and reproductive function in ewes fed tall fescue seed and to determine whether addition of fish meal (FM) to a diet of EI fescue would alter PGF_2α production. Ewes were fed a diet with novel or non-toxic endophyte-infected (NE) or EI tall fescue seed containing either corn gluten meal (CG) or FM. Serum concentrations of prolactin, a measure of severity of fescue toxins, were reduced in ewes fed EI compared with NE forage seed (forage × day, P < 0.02) and were greater in NEFM than NECG-fed ewes (P < 0.03). Size and number of corpora lutea (CL), determined by trans-rectal ultrasonography, were similar between diets (P > 0.10). Serum concentrations of progesterone were reduced in ewes with two CL fed EI compared with NE seed (forage × CL number × day, P < 0.001). Oxytocin-induced PGFM concentrations during the luteal phase were determined as a measure of uterine function. On the day of oxytocin administration, peak plasma concentrations of PGFM were reduced in EI compared with NE-fed ewes (forage × time, P < 0.003), but FM did not influence PGFM concentrations. Estrous cycle length was more variable in EI than NE-fed ewes. There appears to be some asynchrony between NE and EI-fed ewes leading to changes in uterine responsiveness to oxytocin. Inclusion of FM did not alter uterine responsiveness to oxytocin.     

Temporal Relationships Between Oxytocin and 13,14-Dihydro-15-Keto-Prostaglandin F2α Pulses in Ovariectomized Ewes

The primary objective was to evaluate the role of non-ovarian oxytocin in the initiation of pulses of PGF_2α, as measured by peripheral concentrations of 13,14-dihydro-15-keto-prostaglandin F_2α (PGFM). A 2 × 2 factorial arrangement of estradiol and progesterone treatments was administered to groups of five ewes after ovariectomy on Day 12. Progesterone (10 mg) was administered at 0700 and 1900 hr on Day 12, and then either progesterone or its vehicle was administered on Days 13 and 14. Silastic implants, either empty or containing estradiol, was administered at ovariectomy. Oxytocin and PGFM were measured in jugular blood samples withdrawn from an indwelling catheter at 5-min intervals for 8 hr on Day 15. Statistically significant pulses of oxytocin, presumably of posterior pituitary origin, were detected in all ewes. Approximately one-half of the oxytocin pulses preceded a pulse in PGFM concentrations by 10 min or less. These pulses tended (P = 0.09) to have a longer duration than those not linked to pulses of PGFM. The number of PGFM pulses that followed or did not follow an oxytocin pulse by 10 min or less was similar (P > 0.2). The amplitude and duration of oxytocin-linked PGFM pulses were greater (P = 0.05) than non-linked pulses. Although several explanations for the lower than anticipated temporal relationship between oxytocin and PGFM pulses are possible, the finding that oxytocin-related PGFM pulses are distinguishable from other pulses is consistent with the concept that oxytocin initiates robust pulses in PGF_2α secretion.     

Effects of suckling, progestogen-impregnated pessaries or hysterectomy on ovarian function in autumn-lambing postpartum ewes

In three experiments, we examined the effects of suckling, progestogen treatment, hysterectomy or exogenous gonadotropin releasing hormone (GnRH) on ovarian function in autumn-lambing, postpartum ewes. In each experiment, GnRH was injected on approximately d 25 postpartum. Suckling reduced (P less than .01) GnRH-induced release of luteinizing hormone (LH) but not of follicle stimulating hormone (FSH), and reduced (P less than .05) the proportion of ewes that developed corpora lutea in response to GnRH. Suckling had no effect on duration (8.8 d) of GnRH-induced luteal phases. Progestogen prior to GnRH increased (P less than .01) the duration of the first luteal phase (10.1 vs 7.6 d; progestogen-treated ewes vs control ewes), but progestogen did not affect the release of LH or FSH. Progestogen treatment did not alter the interval from parturition to the first detected estrus (42.6 d). The concentration of 13,14-dihydro-15-keto-PGF2 alpha (PGFM) just after lambing was greater than 400 pg/ml of jugular plasma, but concentrations of PGFM declined thereafter. Hysterectomy the day after lambing hastened (P less than .001) the decline in concentrations of PGFM, indicating that prostaglandins from the postpartum uterus probably caused the high concentrations of PGFM in jugular plasma. Hysterectomy reduced (P less than .05) the interval from parturition to detectable luteal function (19.6 vs 25.3 d) and enhanced (P less than .001) luteal production of progesterone. This study of autumn-lambing ewes indicates that the uterus has a negative effect on ovarian function and that suckling and progestogen affect ovarian response to GnRH.     

Effect of repeated administration of oxytocin during diestrus on duration of function of corpora lutea in mares

OBJECTIVE: To determine whether IM administration of exogenous oxytocin twice daily on days 7 to 14 after ovulation blocks luteolysis and causes prolonged function of corpora lutea (CL) in mares. DESIGN: Prospective study. ANIMALS: 12 mares. PROCEDURES: Beginning on the day of ovulation (day 0), jugular blood samples were collected every other day until day 40 for determination of progesterone concentration. On day 7, mares (n = 6/group) were treated with saline (0.9% NaCl) solution (control group) or oxytocin. Beginning on day 7, control mares received 3 mL of sterile saline solution every 12 hours, IM, and oxytocin-treated mares received 60 units of oxytocin every 12 hours, IM, through day 14. Mares were considered to have prolonged CL function if progesterone concentration remained > 1.0 ng/mL continuously through day 30. RESULTS: The proportion of mares with prolonged CL function was significantly higher in the oxytocin-treated group (6/6), compared with the control group (0/6). All control mares underwent luteolysis by day 16, at which time their progesterone concentrations were < 1.0 ng/mL. In contrast, all 6 oxytocin-treated mares maintained progesterone concentrations > 1.0 ng/mL continuously through day 30. CONCLUSIONS AND CLINICAL RELEVANCE: IM administration of 60 units of oxytocin twice daily on days 7 to 14 after ovulation was an efficacious method of inhibiting luteolysis and extending CL function in mares. Disrupting luteolysis by administering exogenous oxytocin during diestrus appears to be a plausible and practical method of long-term suppression of estrus in mares.     

Effects of estradiol-17 beta, naloxone and gonadotropin releasing hormone on postpartum secretion of luteinizing hormone in fall-lambing ewes

The interaction among exogenous estradiol-17 beta, naloxone and gonadotropin releasing hormone (GnRH) in the control of luteinizing hormone (LH) secretion was studied in intact postpartum ewes nursing their offspring. One-half of 30 fall-lambing ewes were implanted subcutaneously with an estradiol-17 beta containing Silastic capsule between postpartum d 1 and 12 which doubled their serum concentrations of estradiol (16.0 +/- .1 vs 8.4 +/- .1 pg/ml). Blood samples were collected from implanted and non-implanted ewes at 15-min intervals for 5 h on d 3, 8, 13, 20 and 28 postpartum. Pre-injection samples were collected for 1 h, and ewes were injected with saline, naloxone (NAL;1 mg/kg) or GnRH (100 micrograms/ewe). When averaged across all days and implant groups, serum LH in the three post-NAL samples was higher (P less than .05) than in the three pre-NAL samples (3.6 +/- 1.2 vs .6 +/- .2 ng/ml). Post-GnRH concentrations of serum LH were lower (P less than .05) in estradiol-implanted ewes than in non-implanted ewes on d 8 and 13, but there were no differences in any LH characteristics on d 20 and 28 after implant removal on d 12. In non-implanted ewes, serum LH responses to GnRH increased (P less than .05) eightfold from d 3 (3.8 +/- 1.4 ng/ml) to d 8 (31.6 +/- 1.4 ng/ml), remained elevated through d 20, but declined by d 28 (10.8 +/- 1.4 ng/ml).(ABSTRACT TRUNCATED AT 250 WORDS)     

Hormonal, estrual, ovulatory and milk traits in postpartum dairy cows following multiple daily injections of oxytocin

Release of oxytocin at suckling or milking may delay onset of estrous cycles in postpartum cows. Twenty lactating Holsteins of mixed parity were given 100 mU oxytocin iv (n = 10) or 2 ml saline (control; n = 10) via jugular catheters at 0530, 0930, 1730 and 2130 daily from calving (d o) until 28 d postpartum. All cows were milked twice daily at 0130 and 1330. Blood was collected thrice weekly (Monday, Wednesday, Friday at 0530) for 12 wk and analyzed by radioimmunoassay for progesterone and 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM) in serum. On d 12, blood was collected every 15 min for 6 h via jugular catheters and concentrations of luteinizing hormone (LH), cortisol and PGFM were determined. Rate of involution of the reproductive tract was estimated twice weekly by palpation per rectum. Overall mean, baseline concentrations, number of pulses/6 h, and pulse duration of LH on d 12 were similar among treatment groups. However, oxytocin seemed to reduce (P less than .10) pulse amplitude of LH in multiparous cows (.4 +/- .2 vs .8 +/- .1 ng/ml), but not in primiparous cows. Concentrations of cortisol and PGFM in serum on d 12 were unaffected by treatment. The average intervals from calving to first ovulation, based on changes of progesterone in serum and the intervals to first estrus, were similar between treatment groups. Rates of involution of the cervix and uterus also were similar between treatments. Milk yield, percent protein in milk and somatic cell counts did not differ between treatment groups. However, percent fat in milk tended to be higher (P less than .10) in cows given oxytocin than in controls (3.99 +/- .22 vs 3.68 +/- .21). These data indicate that multiple daily injections of oxytocin did not affect: 1) length of anestrus and anovulation in postpartum dairy cows, 2) LH release and 3) rates of cervical and uterine involution.     

Effect of oxytocin on expression of cytosolic phospholipase A2 mRNA and protein in ovine endometrial tissue in vivo

The induction of endometrial prostaglandin (PG) F₂ synthesis by oxytocin is dependent upon activation of phospholipase (PL) A₂ and mobilization of arachidonic acid. The objective of this study was to determine if oxytocin stimulates PGF₂ synthesis by inducing synthesis of cytosolic PLA₂ (cPLA₂). In Experiment 1, 15 ovariectomized ewes were given progesterone and estradiol to simulate an estrous cycle. Ewes were then given an injection of oxytocin on Day 14 of the simulated estrous cycle. Jugular blood samples were collected and assayed for 13,14-dihydro-15-keto-prostaglandin F₂ (PGFM). Uteri were collected at 0, 7.5, 25, 90, or 240 min postinjection (n = 3 ewes/time point). Total RNA was isolated from caruncular endometrium and subjected to dot-blot analysis. Oxytocin induced a rapid and transient increase in serum PGFM (P < 0.01). However, endometrial concentrations of cPLA2 mRNA did not change following oxytocin administration (P > 0.10). In Experiment 2, 11 ovary-intact ewes were given oxytocin (n = 5) or saline (n = 6) on Day 15 after estrus. Jugular blood samples were collected and assayed for serum concentrations of PGFM. Uteri were collected at 15 min postinjection. Homogenates were prepared from caruncular endometrium and subjected to Western blot analysis. Concentrations of PGFM were higher in oxytocin treated ewes compared to saline treated ewes at 15 min postinjection (P < 0.01). Endometrial concentrations of cPLA₂ protein were greater in the cytosolic than in the microsomal fraction (P < 0.01). Oxytocin did not affect the amount of cPLA₂ protein in either fraction (P > 0.10). In conclusion, oxytocin did not effect expression of either cPLA₂ mRNA or protein in ovine endometrium. Oxytocin may stimulate PGF₂ synthesis by activating cPLA₂ protein that is already present in an inactive form.     

Hormonal profiles of late gestation ewes following intra-uterine inoculation with and without lux-modified Escherichia coli

The objectives of these investigations were to develop an ovine model for Escherichia coli (E. coli)-induced preterm delivery, and monitor ewe hormonal response. EXP 1: Ewes (105 +/- 13 days of gestation) were allotted to the following intra-uterine inoculations: Saline-(CON; n=5); 1 x 10(6) CFU/ml (Low Treatment, LT; n=6); or 1 x 10(7) CFU/ml (High Treatment, HT; n=6) E. coli. Twenty-four h after inoculation, the HT ewes had increased (P<0.05) cortisol compared to LT and CON ewes, and HT and LT ewes had increased (P<0.05) progesterone compared to CON ewes. Preterm delivery was 33% for LT ewes and 0% for HT and CON ewes. EXP 2: Ewes (124 +/- 18 days of gestation) were allotted to the following intra-uterine inoculations using lux-modified E. coli: Trial-1: Luria Broth (LB; CT1; n=5); 4.0 x 10(6) CFU (n=5), 20.0 x 10(6) CFU (n=5); and Trial-2: LB (CT2; n=5), 1.2 x 10(6) CFU (n=5), and 5.6 x 10(6) CFU (n=5) E. coli-lux. Preterm delivery occurred between 48 and 120 h post-inoculation in 60, 25, 60 and 75% of ewes infected with 1.2, 4.0, 5.6, and 20 x 10(6) CFU, respectively. Serum cortisol and progesterone did not differ (P>0.05) between CT1 or CT2 and inoculated ewes. In summary, 25 to 75% of ewes inoculated preterm delivered. However, variable results in cortisol and progesterone profiles between Control and inoculated ewes were observed between the two studies.     

Hormonal induction of lactation in non-pregnant ewes

Exogenous hormone treatments designed to induce nonpregnant ewes to lactate at levels sufficient to rear orphan lambs were investigated in three breeds (Blackface, Dorset and Greyface) and in a total of 14 groups. The hormone treatments consisted of an induction phase lasting 1-6 weeks during which most ewes received daily subcutaneous injections of 75 mg of progesterone and 100 microg of oestradiol-17beta, and a trigger phase lasting 1 or 2 weeks during which 5 mg of oestradiol-17beta and/or 10 mg of dexamethasone were given daily and the daily dose of progesterone was reduced to zero. From the end of the trigger phase for at least 2 weeks, milk production was determined three times daily by hand milking after prior intravenous injections of 5 IU of oxytocin. The linear dimensions of the udder were measured in each ewe at 5-6 day intervals throughout the induction and trigger phases. Udder sizes increased in response to hormone treatment in all cases, and the rate of increase was usually 1.75-11.3 times greater during the trigger phase than during the induction phase. Accumulation of milk in the udder during the trigger phase resulted in mean milk yields of 212-763 ml on the first day of milking, which were higher than the mean yields of 130-354 ml on the second day. Thereafter the mean daily milk yields increased progressively to reach 579-1301 ml after 14 days of milking. Group comparisons revealed the following: an induction phase of at least 4 weeks duration was required to ensure that all hormone-treated ewes produced 800 ml or more of milk/day by 14 days of milking; during the trigger phase, oestradiol-17beta alone was a more effective lactogenic agent than dexamethasone alone, and dexamethasone apparently hindered the lactogenic actions of oestradiol-17beta when both hormones were given together; extending the duration of the trigger phase from 1 to 2 weeks did not improve subsequent milk yields; and there were no significant breed differences in milk yield responses to similar hormone treatments. It is concluded that a 4-6 week induction phase followed by a 1-week trigger phase using the progesterone and oestradiol-17beta doses noted above but excluding dexamethasone would induce in most nonpregnant ewes lactation at levels sufficient to rear orphan lambs. Compared to the compositions of normal colostrum and milk, the milk from some of the present ewes had lower dry matter contents, fat concentrations and immunoglobulin-G concentrations and generally similar lactose concentrations. These differences were not considered to be sufficient to jeopardize the survival of lambs reared by such ewes.     

Effect of various veterinary procedures on plasma concentrations of cortisol, luteinising hormone and prostaglandin F2 alpha metabolite in the cow

Five commonly practised veterinary procedures were studied: Palpation per rectum of the reproductive tract, intramuscular injection, single venepuncture, repeated venepuncture and jugular vein catheterisation. Plasma cortisol concentrations increased from baseline values of approximately 2 ng/ml to maximum mean values between 6.5 +/- 2.5 ng/ml and 13.8 +/- 5.6 ng/ml approximately 13 to 27 minutes after each manipulation. Baseline values occurred approximately 80 minutes later. In the control bleeding periods unacclimatised cows initially had high values of plasma cortisol (5 to 10 ng/ml) which returned to baseline after two hours, ie, before beginning any procedure. There were no statistically significant changes in luteinising hormone concentrations. The concentration of 13,14 dihydro, 15-keto prostaglandin F2 alpha (PGFM) increased from 61.0 +/- 4.6 pg/ml to 209.8 +/- 152.1 pg/ml in three out of five cows palpated on days 16 and 19 of the oestrous cycle. Increases did not occur in five other cows palpated during the follicular phase, nor in five cows palpated on day 12. However, after palpation on day 8, one animal did have concentrations of PGFM similar to those occurring during spontaneous release on days 18 to 20 of the oestrous cycle.     

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.     

Progesterone and the surge release of gonadotropins in the ewe

On day 12 of an estrous cycle, 4 groups of ewes were treated with either blank (no steroid) or 3 different sizes of progesterone-containing rubber implants to study the effect of maintained progesterone levels on preovulatory events. Following luteolysis progesterone levels were 0.55 ± 0.13 ng/ml in control ewes and 0.62 ± 0.10 ng/ml, 0.99 ± 0.09 ng/ml and 1.85 ± 0.04 ng/ml in the 3 groups of progesterone-treated ewes. Preovulatory surges of LH and FSH occurred in $\text{5}\text{5}\text{,}\text{5}\text{5}\text{;}\text{4}\text{5}\text{,}\text{3}\text{5}\text{;}\text{4}\text{5}\text{,}\text{3}\text{5}$ and $\text{0}\text{5}\text{,}\text{0}\text{5}$ ewes in these 4 groups respectively. Eleven days after implant insertion, all ewes responded to a GnRH challenge. The height of all FSH peaks was depressed by progesterone treatment (P<0.05). Three groups of ewes were ovariectomized at day 6 of a cycle and treated with estradiol-17β and progesterone-containing implants. After 8 days of treatment, progesterone implants were removed in sections to give 3 different rates of decline in serum progesterone levels. The gonadotropin surges occurring following progesterone removal were delayed by the slower rates of progesterone decline. Another group of ewes was treated as in experiment 2, but the progesterone implants were all removed together after 8 days. Subsequent replacement of progesterone implants, after 12 or 18 hours, blocked the gonadotropin surges in all, or 2 of 6 ewes respectively. When replaced after 24 hours, implants producing low progesterone (1.38 ± 0.22 ng/ml) did not block gonadotropin surges, but in 2 of 5 ewes high progesterone did (2.87 ± 0.22 ng/ml). Removal of progesterone implants, 12 or 24 hr after replacement, produced secondary gonadotropin surges of smaller magnitude than the initial peaks (P<0.01).     

Administration of single short-acting doses of GnRH antagonist modifies pituitary and follicular function in sheep

Current study determined the effect of two different single subcutaneous doses (1.5 and 3 mg) of GnRH antagonist (GnRHa) on pituitary and follicular function in non-lactating cyclic ewes. Both doses abolished the pulsatile secretion of luteinizing hormone (LH) for at least 3 days and decreased mean LH concentration during 6 days (0.64 +/- 0.09 for control and 0.54 +/- 0.05, P < 0.005, and 0.46 +/- 0.02, P < 0.00001, for 1.5 and 3 mg, respectively). Supply of GnRHa decreased the number of large dominant follicles, so the total number of smaller follicles, 2-3 mm in size, increased in both treated groups from day 0, reaching its maximum at day 2 in ewes treated with 1.5 mg (19.83+/-1.05 versus 5.83 +/- 0.50 in the control, P < 0.005) and at day 4 in sheep treated with 3mg (18.67 +/- 0.65 versus 5.50 +/- 0.65 in the control, P < 0.0001). However, the analysis of follicular function in terms of inhibin A indicated a possible effect of the higher dose of GnRHa on follicular function. The pattern of inhibin secretion in the group treated with 3mg of GnRHa decreased after the first 48 h, reaching its lowest value on day 4.5 (182.59 +/- 3.75 to 140.28 +/- 9.91 pg/ml, P < 0.05) concentration significant lower than control sheep (171.93 +/- 6.21 pg/ml, P +/- 0.01) or treated with 1.5 mg (168.04 +/- 7.16 pg/ml, P+ /- 0.05). Hence, the use of 1.5 mg would be more suitable to induce the presence of a high number of follicles able to grow to preovulatory sizes.     

Delayed luteolysis and suppression of the pulsatile release of oxytocin after indomethacin treatment in the goat

Luteolysis in the goat was characterised by the pulsatile appearance of both oxytocin and 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM), the primary pulmonary metabolite of prostaglandin (PG) F2 alpha, in the peripheral circulation. The episodic, surge release of oxytocin was superimposed on declining levels which paralleled the fall in progesterone concentrations. Daily subcutaneous administration of the prostaglandin synthetase inhibitor indomethacin (10 mg kg-1) between days 11 and 16, delayed luteolysis and suppressed both the decline in oxytocin concentrations and the pulsatile appearance of both oxytocin and PGFM in the peripheral circulation. Although there was little evidence of the release of the two hormones being synchronised, the results suggest that PGF2 alpha may stimulate pulsatile oxytocin release at luteolysis.     

Involvement of beta-adrenoceptors in the regulation of luteal function in cattle.

The infusion of beta-adrenomimetic drug (bamethan sulphate--BS) on day 11 of the oestrous cycle markedly increased peripheral progesterone concentration whereas infusion of beta-antagonist (propranolol) decreased plasma progesterone levels in heifers. Oxytocin injected on days 2-6 of the oestrous cycle shortened the length of the cycle (to 7-9 days) whereas the length of the cycle in heifers injected with oxytocin followed by injections of either BS or xanthinol (theophylline nicotinate--a potent vasodilatory drug) was normal (21.8 +/- 1.7). Heifers injected with BS on days 13-21 of the oestrous cycle had increased levels of progesterone on days 13-19 of the cycle and the length of the cycle was normal (22.3 +/- 0.6 compared with 21.3 +/- 1.5 for the control). The data indicate that: (1) beta-adrenergic stimulation overcomes the action of oxytocin given on days 2-6 of the cycle, whereas the stimulation of beta-receptors on days 13-21 of the oestrous cycle raised progesterone plasma concentration but did not prevent spontaneous luteolysis. (2) Basal secretion of progesterone in cattle is partly dependent on constant beta-adrenergic stimulation.