Interrelationships between progesterone, 13,14-dihydro-15-keto PGF-2 alpha (PGFM) and LH in cyclic and early pregnant cows

Plasma progesterone and LH secretion patterns were examined in 18 mature dairy cows during the oestrous cycle and after insemination. Blood samples were collected every 15 min for 8 h per day on Days 3, 5, 6, 7, 8, 9, 10, 12, 14, 16, 17, 18, 19, 20 and 21 of the oestrous cycle, then, in the same cows, at the same times during early pregnancy. PGF-2 alpha secretion rates (as determined by plasma PGFM concentrations) were also monitored on Days 14, 16 and the day of, or equivalent to, luteal regression. Mean daily plasma progesterone concentrations were similar until Day 16 in cyclic and pregnant cows, after which values in non-pregnant animals declined. Regression analysis indicated that progesterone concentrations were best described by a quadratic expression with fitted maximum values on Day 13 in non-pregnant animals but values increased linearly over the whole period to Day 21 in pregnant cows. The frequency, amplitude and area under the curve of LH episodes showed no significant differences between cyclic and pregnant animals. In pregnant cows, the amplitude and area under the curve of progesterone episodes increased linearly between Days 8 and 21, although no such increase occurred in cyclic cows. Low-level PGFM episodes were present in cyclic and pregnant cows on Days 14 and 16 after oestrus, and high amplitude episodes occurred in non-pregnant cows during luteal regression. Pregnant cows showed a significant depression of the amplitude, but not the frequency of episodes at the expected time of luteal regression. These results confirm that the corpus luteum of pregnancy secretes an increasing amount of progesterone per se and per unit of LH until at least Day 21 after mating. They further suggest that the corpus luteum of the cyclic cow may experience small episodes of PGF-2 alpha and be subjected to initial degenerative changes by Day 14 after oestrus, some time before the onset of definitive luteolysis.     

Characteristics of prostaglandin F measurements in the ovarian circulation during the oestrous cycle and early pregnancy in the cow

Holstein or crossbred beef cows were anaesthetized on Days 15 to 17 after oestrus; the ovarian artery (OA), ovarian (utero-ovarian) vein (OV) and a peripheral artery (PA), were catheterized for chronic blood sampling. Beginning on the day after surgery, 6 sequential blood samples were collected every 30-40 min twice daily from 8 cyclic and 6 pregnant cows during Days 16-20: 818 blood samples (including 216 OA and PA concurrent arterial pairs) were collected. Overall least squares means for PGF concentrations (pg/ml) in the OV, OA and PA of cyclic cows were 562, 228 and 106, respectively. A significant (P less than 0.01) OA-PA difference (122 pg/ml) suggests that a local transfer system, between uterine venous effluent and ovarian arterial affluent, is functional in the cow. A transfer efficiency of about 1% was estimated. In cyclic cows differences in OA-PA concentrations of PGF were minimal on Days 16-18 and increased to about 160 pg/ml during luteal regression (Days 19-20). In pregnant cows a biphasic OA-PA pattern of difference in PGF between days was detected, with a peak on Day 18 (136 pg/ml) which was not apparent on Days 19-20. Amplitude of PGF spikes in the OA was significantly higher in cyclic (725 pg/ml) than in pregnant cows (397 pg/ml). We suggest that pregnancy suppresses PGF delivery to the ovarian circulation, resulting in maintenance of the corpus luteum in pregnant cows.     

A morphometric analysis of the corpus luteum of the cow during the estrous cycle and early pregnancy

Corpora lutea were collected from cows on Days 6, 8, 10, 12, 14, 16, 18 and 19 of the estrous cycle and early pregnancy (n=2/d) and were examined by light microscopy. Mean lutein cell diameter was significantly (P<0.05) greater in pregnant than in cyclic cows on Days 6, 8, 10, 12, 16, 18 and 19 (cyclic versus pregnant: Day 6: 13.9 +/- 0.22 vs 14.9 +/- 0.24; Day 8: 13.8 +/- 0.20 vs 15.4 +/- 0.2; Day 10: 14.8 +/- 0.24 vs 17.4 +/- 0.24; Day 12: 13.2 +/-0.25 vs 17.9 +/- 0.31; Day 16: 13.9 +/- 0.28 vs 16.5 +/- 0.31; Day 18: 13.0 +/- 0.22 vs 16.5 +/- 09.36, and Day 19: 15.0 +/- 0.23 vs 17.6 +/- 0.33 mum, respectively). The distribution of cell sizes was leptokurtotic throughout the estrous cycle and the first 10 d of pregnancy, but tended towards bimodality after Day 14 of pregnancy. The proportion of lutein cell cytoplasm occupied by vacuoles was lower in pregnant than in cyclic cows from the 12th day post estrus, but there was a marked (P<0.05) increase in vacuolation of cells from cows undergoing luteolysis. Stainable intercellular collagen was also less abundant in pregnant than cyclic cows from the 12th day post estrus. The higher rate of progesterone secretion of pregnant, compared with cyclic cows may be attributed to the greater numbers and greater contribution to luteal mass of large lutein cells in the corpus luteum of pregnancy.     

Hormonal changes during luteal regression in farmed fallow deer, Dama dama

Concentrations of progesterone, oxytocin and PGFM (pulmonary metabolite of PGF-2 alpha) were measured in plasma from peripheral blood samples collected from 5 fallow does every hour or 2 h for 12-h periods on Days 15-20 inclusive of the oestrous cycle (i.e. luteolysis). For 3 does that exhibited oestrus on Day 21, plasma progesterone concentrations fluctuated between 3 and 10 ng/ml on Days 15-18 inclusive. Thereafter, values declined progressively to attain minimum concentrations of less than 0.05 ng/ml on Day 20. Basal concentrations of plasma oxytocin and PGFM fluctuated between 5 and 20 pg/ml and 10 and 100 pg/ml respectively. Episodic pulses of plasma oxytocin (greater than 300 pg/ml) occurred on Days 15 and 16, whereas pulses of plasma PGFM (greater than 400 pg/ml) occurred on Days 19 and 20. There was little apparent correlation between episodic pulses of the two hormones. For 2 does that exhibited oestrus on Day 22, plasma progesterone concentrations declined to minimum values of 1.0-1.5 ng/ml by Day 20. One of these does showed very high levels of oxytocin secretion throughout the sampling period while the other showed an apparent paucity of oxytocin secretory periods. Two does hysterectomized on Day 13 of their second oestrous cycle failed to exhibit further oestrous cycles. Continual elevation of plasma progesterone concentrations (2-6 ng/ml) for an 8-month period indicated persistence of the corpus luteum after hysterectomy. It is concluded that luteolysis in fallow deer involves episodic secretion of both oxytocin and PGF-2 alpha.     

Uterine blood flow of cows during the oestrous cycle and early pregnancy: effect of the conceptus on the uterine blood supply.

Blood flow to each uterine horn of cows during the oestrous cycle and early pregnancy was determined daily by use of electromagnetic blood flow probes placed around both middle uterine arteries. The pattern of blood flow to uteri of pregnant and non-pregnant cows was similar until Day 14 after mating or oestrus. Between Days 14 and 18 of pregnancy blood flow to the uterine horn containing the conceptus increased (P less than 0.01) 2- to 3-fold, whereas blood flow to the non-gravid uterine horn in these cows remained constant. No corresponding increase in blood flow to the uterine horn ipsilateral to the ovary bearing the CL was observed in non-pregnant cows during this 4-day period. By Day 19 of pregnancy, blood flow to the gravid uterine horn had returned to a level similar to that observed on Day 13. Blood flow to both uterine horns of pregnant cows remained constant from Days 19 to 25 and then increased to the gravid horn (P less than 0.01) markedly until Day 30 whereas blood flow to the non-gravid horn remained low. Uterine blood flow during the oestrous cycle of non-pregnant cows was positively correlated (P less than 0.01) with systemic concentrations of oestradiol and the ratio of oestradiol (pg/ml) to progesterone (ng/ml). There was no association between oestradiol concentrations and blood flow to the gravid uterine horn. These data indicate local control of uterine blood flow by the bovine conceptus which may function to create optimal conditions for the continuation of pregnancy.     

Myometrial activity and plasma progesterone and oxytocin concentrations in cycling and early-pregnant ewes

Myometrial activity and plasma progesterone (P) and oxytocin (OT) were measured in early pregnant (n = 5) and cycling (n = 5) ewes. Electromyography (EMG) leads and jugular and inferior vena cava (IVC) catheters were surgically placed in ewes about 1 wk before data collection. When ewes returned to estrus, they were bred to either an intact or vasectomized ram. Continuous EMG data were collected, and blood samples were collected twice daily from day of estrus (Day 0) until Day 18. Ewes bred with an intact ram were checked surgically for pregnancy on Day 20. Computerized, quantitative analysis of EMG events showed no difference in signal from the right to left uterine horns, and no differences between pregnant and cycling ewes (p less than 0.05) until Days 14-18 when nonpregnant ewes returned to estrus and had increased EMG activity. The mean number of EMG events 180-900 s in length decreased in pregnant ewes, but this difference was not significant (p less than 0.05). Jugular plasma progesterone (P) levels confirmed corpus luteum (CL) formation in all ewes, and no differences in P between pregnant and nonpregnant ewes were measured until Days 14-18, when cycling ewes underwent luteolysis and pregnant ewes maintained CL. IVC plasma oxytocin concentrations were increased in pregnant ewes compared to concentrations in nonpregnant ewes on Days 5-13 (p less than 0.05), and the difference was largest at Day 6 (means +/- SEM pg/ml: pregnant = 68.7 +/- 13.9, nonpregnant = 30.9 +/- 19.9).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of an agonist of gonadotrophin releasing hormone in cattle. I. Hormone concentrations and oestrous cycle length

Four trials were completed to study the effects of a single intramuscular injection of 5 μg of an agonist of gonadotrophin releasing hormone (Hoe 766) on plasma concentrations of LH and progesterone, and on oestrous cycle length in normally cycling dairy cows.The first trial (four cows) showed that a mid-cycle injection of Hoe 766 temporarily increases plasma LH from less than 5 ng/ml to over 20 ng/ml within 2.5 h. Average plasma progesterone concentrations ranged from 4.8 to 7.0 ng/ml compared to 3.3 ng/ml in the control animal.The second trial (22 cows) showed that an injection of Hoe 766 on Cycle Day 3, 6 or 9 (Oestrus = Cycle Day 0) increased average plasma progesterone concentrations during Cycle Days 13, 14 and 15 by 1.2 ng/ml. Each of three cows injected on Cycle Day 16 maintained plasma concentrations above 3.9 ng/ml until Cycle Day 19 and corpus luteum (CL) size was maintained until Cycle Day 21. Except for the group of cows injected on Cycle Day 3, all other groups had temporarily reduced concentrations of plasma progesterone when sampled 24 h after Hoe 766 administration.The third trial (216 cows) showed that a single injection of Hoe 766 made between Cycle Day 1 and 10 did not alter oestrous cycle length (21.5 vs 21.3 days). In contrast, in the fourth trial (371 cows), a single injection of Hoe 766 between Cycle Days 12 to 16 altered the distribution of cycle lengths of 17–29 days, the average cycle length and the incidence of ovulation without detected oestrus. Compared to matched control cows, fewer Hoe 766-treated cows were detected in oestrus (73.9% vs 90%), or had cycle lengths of less than 20 days (4.7% vs 22.2%). These effects were most pronounced among cows injected on Cycle Day 16 when only 51.7% were detected in oestrus and their average cycle length was 24.1 days.These effects were not due to the formation of a secondary CL. Rather, the injection of Hoe 766 stimulated CL function and appeared to prevent or delay normal luteolysis when administered from Cycle Day 12.     

Extension of oestrous cycles and prolonged secretion of progesterone in non-pregnant cattle infused continuously with oxytocin

The experimental objective was to evaluate how continuous infusion of oxytocin during the anticipated period of luteolysis in cattle would influence secretion of progesterone, oestradiol and 13,14-dihydro-15-keto-prostaglandin F-2 alpha (PGFM). In Exp. I, 6 non-lactating Holstein cows were infused with saline or oxytocin (20 IU/h, i.v.) from Day 13 to Day 20 of an oestrous cycle in a cross-over experimental design (Day 0 = oestrus). During saline cycles, concentrations of progesterone decreased from 11.0 +/- 2.0 ng/ml on Day 14 to 2.0 +/- 1.3 ng/ml on Day 23; however, during oxytocin cycles, luteolysis was delayed and progesterone secretion remained near 11 ng/ml until after Day 22 (P less than 0.05). Interoestrous interval was 1.6 days longer in oxytocin than in saline cycles (P = 0.07). Baseline PGFM and amplitude and frequency of PGFM peaks in blood samples collected hourly on Day 18 did not differ between saline and oxytocin cycles. In Exp. II, 7 non-lactating Holstein cows were infused with saline or oxytocin from Day 13 to Day 25 after oestrus in a cross-over experimental design. Secretion of progesterone decreased from 6.8 +/- 0.7 ng/ml on Day 16 to less than 2 ng/ml on Day 22 of saline cycles; however, during oxytocin cycles, luteolysis did not occur until after Day 25 (P less than 0.05). Interoestrous interval was 5.9 days longer for oxytocin than for saline cycles (P less than 0.05). In blood samples taken every 2 h from Day 17 to Day 23, PGFM peak amplitude was higher (P less than 0.05) in saline (142.1 +/- 25.1 pg/ml) than in oxytocin cycles (109.8 +/- 15.2 pg/ml). Nevertheless, pulsatile secretion of PGFM was detected during 6 of 7 oxytocin cycles. In both experiments, the anticipated rise in serum oestradiol concentrations before oestrus, around Days 18-20, was observed during saline cycles, but during oxytocin cycles, concentrations of oestradiol remained at basal levels until after oxytocin infusion was discontinued. We concluded that continuous infusion of oxytocin caused extended oestrous cycles, prolonged the secretion of progesterone, and reduced the amplitude of PGFM pulses. Moreover, when oxytocin was infused, pulsatile secretion of PGFM was not abolished, but oestrogen secretion did not increase until oxytocin infusion stopped.     

Uterine oxytocin receptors in cyclic and pregnant cows.

Binding of [3H]oxytocin to uterine subcellular preparations ('oxytocin receptor concentrations') was measured in uterine tissue of heifers and multiparous dairy cows at various stages of the oestrous cycle and during early pregnancy. A method for the assay of ovine uterine oxytocin receptors was optimized for use on bovine tissue. Oxytocin receptor concentrations were increased in cyclic animals around the period of luteolysis and oestrus, rising on Day 15 in endometrium and on Day 17 in myometrium while pregnant animals showed no comparable rise. Receptor concentrations then declined on Day 3 after oestrus in myometrium and on Day 5 in endometrium. Some cyclic animals did not show the expected rise in receptors in the late luteal phase; these animals had abnormally high progesterone concentrations for this stage of the cycle. In animals slaughtered on Day 18 after oestrus and/or insemination which had low oxytocin receptor levels, plasma progesterone concentrations were consistently high; while all animals showing the late luteal phase elevation in receptor values had low progesterone concentrations. Oxytocin receptor and progesterone concentrations were negatively correlated (P less than 0.05). These data support the hypothesis that oxytocin receptor level is a key factor in the process of luteolysis in cattle and that in pregnancy there is suppression of uterine oxytocin receptor at the expected time of luteolysis. We suggest that uterine oxytocin receptor levels are partly controlled by circulating steroid hormones and are suppressed during early pregnancy.     

Characterization of the uterine environment during early conceptus expansion in the bovine

Fifty-three Angus and Hereford beef cows were utilized to investigate the effect of the conceptus on uterine environment during the period of pregnancy recognition. Blood samples were collected on Days 10, 12, 14, 16 and 18. Cows were randomly assigned to be either mated on the subsequent oestrus or serve as cyclic controls. Blood samples were then collected daily from Day 10 until slaughter on Day 15, 16 or 17 from the initiation of oestrus. Uteri were flushed with physiological saline and flushings analyzed for quantitative and qualitative protein changes, calcium, oestradiol-17β and prostaglandin F content. Endometrial explants of caruncular and intercaruncular tissue, and conceptus tissue recovered from pregnant cows were cultured with [³H]-leucine to determine quantitative and qualitative polypeptide synthesis and release. Plasma progesterone concentrations were similar between pregnant and cyclic cows from Day 10 through 17. Only the uterine content of prostaglandin F significantly increased in the ipsilateral horn of pregnant cows on Days 16 and 17. This increase in prostaglandin content was related to the increase in conceptus length from 25 to 40–80 mm. Conceptus production of bovine trophoblastic protein-1 was also first clearly detectable in fluorographs of medium from conceptuses measuring 25 mm. The complexity of the polypeptides present in the medium increased with conceptus development. Polypeptide synthesis by the endometrium was similar between tissues and days; however, production of two groups of low molecular weight basic polypeptides continued to be intensified on fluorographs from the pregnant horn on Day 17 compared to cyclic cows.     

Prostaglandin secretion by perifused bovine endometrium: secretion towards the myometrial and luminal sides at day 17 post-estrus as altered by pregnancy

Bilateral perifusion devices were utilized to measure prostaglandin secretion towards luminal and myometrial sides of bovine endometria. Tissues were collected at Day 17 post-estrus from cyclic (n = 4), pregnant (n = 5) and bred but subsequently non-pregnant (n = 6) cows. Tissue from each cow was placed into two perifusion devices, perifused with Krebs-Ringer Bicarbonate solution (3 ml/10 min) for 2.5 h and fractions collected every 10 min. Oxytocin (1 IU/ml) was perifused during fractions 7-12 to the luminal side of one device and to the myometrial side of the other device. Regardless of status, prostaglandin secretion rates (PGF and PGE2) were higher (P less than 0.01) from the luminal side than the myometrial side. Secretion rates of PGF were lower (P less than 0.01) for endometria from pregnant cows than for endometria from cyclic or bred/non-pregnant cows, whereas secretion rates of PGE2 were not affected by pregnancy status. Regardless of the side of perifusion, secretion rates of PGF and PGE2 from endometria of cyclic and bred/non-pregnant cows were elevated (P less than 0.01) throughout the period of oxytocin treatment, whereas prostaglandin secretion by endometria from pregnant cows was not stimulated by oxytocin. Decreased secretion of PGF from endometria of pregnant cows suggests that the corpus luteum and pregnancy are maintained because of an inhibition of endometrial prostaglandin synthesis or an inability to respond to stimulators of prostaglandin synthesis (i.e. oxytocin).     

Immunosuppressive activity associated with early pregnancy in the bovine

Immunosuppressive activity was assessed in uterine flushings (UF) and uterine vein serum and plasma from nonpregnant and early-pregnant cows, and in media from the short-term culture of Day 18 bovine embryos. The preparations were tested for their ability to inhibit [3H] thymidine (3H-TdR) incorporation into phytohemagglutinin-stimulated bovine lymphocytes. On Days 2-3 (called Day 3), Days 9-10 (called Day 10), and Days 17-19 (called Day 18) of the estrous cycle (estrus = Day 0) and pregnancy, untreated and superovulated cows were anesthesized and jugular vein and uterine vein blood was collected. The uteri were removed and flushed to obtain UF and embryos. Uterine flushings were concentrated and tested for immunosuppressive activity at 400 micrograms uterine protein/ml culture fluid. Uterine flushings from both Day 18 pregnant and Day 18 nonpregnant cows were immunosuppressive (8/8), whereas Day 10 UF were usually not immunosuppressive (7/10). Day 3 UF were usually stimulatory or only marginally suppressive (8/8). Uterine vein serum and plasma from Day 18 cows were not suppressive when compared to jugular vein serum or plasma from the same cow; neither were Day 18 uterine vein serum or plasma suppressive when compared to those same samples taken from Day 3 cows. Embryo culture media obtained from the 48-h culture of Day 18 embryos was consistently suppressive. The activity was lost after dialysis in 1000-Mr cut-off tubing, removed by charcoal, and reduced by protease digestion. These results suggest two mechanisms whereby the embryo could escape immune rejection: 1) the progesterone-induced secretion of a uterine immunosuppressive substance(s) and 2) the production by the embryo of a low molecular weight immunosuppressive substance(s).     

Lack of stimulation of relaxin secretion in lactating sows by suckling in vivo or by oxytocin in vitro.

After parturition, eight sows were zero weaned by removing all piglets 6 h after birth; a further 18 sows suckled at least ten piglets each. Blood samples were collected on Day 4 after zero weaning or on Days 4, 14 and 21 of lactation and the sampling frequency increased during suckling bouts. Ovaries were recovered from sows on these days and corpora lutea were either extracted for estimation of relaxin and progesterone concentration, fixed for immunohistochemical analysis or incubated in vitro in the presence or absence of luteinizing hormone (LH) or oxytocin. Luteal weight and progesterone were higher in the zero-weaned sows than in lactating sows (P less than 0.05 and less than 0.001, respectively); relaxin content was below detection by Day 14. This was supported by immunohistochemical staining for relaxin, which showed limited immunostaining in zero-weaned and Day 4 sows, but none in the tissue recovered on Days 14 and 21, which showed typical signs of regression. Secretion of progesterone and relaxin by luteal tissue in vitro was highest in zero-weaned sows (P less than 0.05), decreased as lactation progressed and neither LH nor oxytocin had any significant effect. Concentrations of plasma relaxin were all less than 0.2 ng/ml in three of the four zero-weaned and Day-4-suckled sows assayed; there was no detectable increase during suckling bouts. It was concluded that during lactation the old corpus luteum of pregnancy is not able to release relaxin in response to suckling in vivo or to oxytocin treatment in vitro.     

Effect of continuous infusion of oxytocin on length of the oestrous cycle and luteolysis in cattle

In Exp. I oxytocin (60 micrograms/100 kg/day) was infused into the jugular vein of 3 heifers on Days 14-22, 15-18 and 16-19 of the oestrous cycle respectively. In Exp. II 5 heifers were infused with 12 micrograms oxytocin/100 kg/day from Day 15 of the oestrous cycle until clear signs of oestrus. Blood samples were taken from the contralateral jugular vein at 2-h intervals from the start of the infusion. The oestrous cycle before and after treatment served as the controls for each animal. Blood samples were taken less frequently during the control cycles. In Exp. III 3 heifers were infused with 12 micrograms oxytocin/100 kg/day for 50 h before expected oestrus and slaughtered 30-40 min after the end of infusion for determination of oxytocin receptor amounts in the endometrium. Three other heifers slaughtered at the same days of the cycle served as controls. Peripheral concentrations of oxytocin during infusion ranged between 155 and 641 pg/ml in Exp. I and 18 and 25 pg/ml in Exp. II. In 4 our of 8 heifers of Exps I and II, one high pulse of 15-keto-13,14-dihydro-prostaglandin F-2 alpha (PGFM) appeared soon after the start of oxytocin infusion followed by some irregular pulses. The first PGFM pulse was accompanied by a transient (10-14 h) decrease of blood progesterone concentration. High regular pulses of PGFM in all heifers examined were measured between Days 17 and 19 during spontaneous luteolysis. No change in length of the oestrous cycle or secretion patterns of progesterone, PGFM and LH was observed.(ABSTRACT TRUNCATED AT 250 WORDS)     

Uterine secretion of prostaglandin F2 alpha in response to oxytocin in ewes: changes during the estrous cycle and early pregnancy.

Experiment 1 was conducted to determine when the ovine uterus develops the ability to secrete prostaglandin F2 alpha (PGF2 alpha) in response to oxytocin and how development is affected by pregnancy. Pregnant and nonpregnant ewes received an injection of oxytocin (10 IU, i.v.) on Day 10, 13, or 16 postestrus. Jugular venous blood samples were collected for 2 h after injection for quantification of 13,14-dihydro-15-keto-PGF2 alpha (PGFM). In nonpregnant ewes, concentrations of PGFM increased following oxytocin on Day 16 but not on Day 10 or 13. Concentrations of PGFM did not increase following treatment on Day 10, 13, or 16 in pregnant ewes. Therefore, the ability of oxytocin to induce uterine secretion of PGF2 alpha develops after Day 13 in nonpregnant but not in pregnant ewes. Experiment 2 was conducted to precisely define when uterine secretory responsiveness to oxytocin develops. Pregnant and nonpregnant ewes received oxytocin on Day 12, 13, 14, or 15. In nonpregnant ewes, concentrations of PGFM increased following treatment on Days 14 and 15, but not earlier. Peripheral concentrations of progesterone showed that uterine secretory responsiveness to oxytocin developed prior to the onset of luteal regression. As in experiment 1, the increase in concentrations of PGFM following administration of oxytocin was much lower in pregnant than in nonpregnant ewes; however, some pregnant ewes did respond to oxytocin with an increase in PGFM. In experiment 3, pregnant ewes received an injection of oxytocin on Day 18, 24, or 30 postmating. Concentrations of PGFM increased following oxytocin on Days 18 and 24. The conceptus appears to delay and attenuate the development of uterine secretory responsiveness to oxytocin.     

Effect of diethylstilboestrol on the relationship between LH, PMSG and progesterone during pregnancy in the mare.

Two studies were conducted to determine the relationship between LH and progesterone and between PMSG and progesterone during pregnancy in mares. In the first, samples of jugular blood were collected daily from 7 mares from the first day of oestrus until Day 28 of pregnancy, and in the second, samples were collected weekly from 14 mares from Day 35 of gestation until parturition. In an attempt to prolong secretion of progesterone from accessory corpora lutea, 7 of these 14 mares were injected with increasing doses (2--10 mg) of diethylstilboestrol (DES) between Days 84 and 142 of gestation. The remaining 7 mares received injections of vehicle. Concentrations of LH, PMSG and progesterone in serum were determined by radioimmunoassay. From the onset of oestrus until Day 4 of gestation, serum concentrations of LH and progesterone were negatively correlated (r = 0.67, P less than 0.01), but from Days 5 to 28 a positive correlation (r = 0.80, P less than 0.01) was noted. Likewise, serum concentrations of PMSG and progesterone were highly correlated between Days 35 and 196 in mares injected with DES (r = 0.72, P less than 0.01) and the vehicle (r = 0.75, P less than 0.01). Injections of DES did not influence serum concentrations of LH, PMSG or progesterone, or affect the length of gestation. It was concluded that DES does not influence the maintenance of pregnancy in the mare.     

Relationship between plasma progesterone concentrations and pregnancy rates in cattle receiving either fresh or previously frozen embryos

Blood samples were collected for the measurement of progesterone concentrations from 320 Holstein-Friesian heifers on Days 7 and 21 post-estrus. All animals were the recipients of either a fresh or previously frozen embryo on Day 7 and were palpated for pregnancy on Day 60 post-estrus. At the time of transer, progesterone levels were highly variable and were not strongly related to subsequent pregnancy status. There was a tendency for lower pregnancy rates in heifers receiving fresh embryos if progesterone levels were less than 1 ng/ml (33 vs 64% overall), and for previously frozen embryos when progesterone concentrations were less than 3 ng/ml (34 vs 44% overall). Progesterone concentrations were not related to subjective evaluation of corpus luteum quality by palpation per rectum. No heifers which maintained pregnancy had progesterone levels less than 1 ng/ml on Day 21. Only 41% of 247 heifers receiving either fresh or previously frozen embryos that were not pregnant on Day 60 had progesterone concentrations less than 1 ng/ml on Day 21. These data suggest that many recipients that do not maintain a pregnancy will experience an extended estrous cycle after transfer.     

Luteal blood flow increases during the first three weeks of pregnancy in lactating dairy cows

The objectives of this experiment were to characterize luteal blood flow in pregnant and non-pregnant cows and to determine its value for early pregnancy diagnosis. Lactating dairy cows (n = 54), 5.2 ± 0.2 y old (mean ± SEM), average parity 2.4 ± 0.2, and ≥ 6 wk postpartum at the start of the study, were used. The corpus luteum (CL) was examined with transrectal color Doppler ultrasonography (10.0-MHz linear-array transducer) on Days 3, 6, 9, 11, 13, 15, 18, and 21 of the estrus cycle (estrus = Day 0). Artificially inseminated cows (n = 40) were retrospectively classified as pregnant (embryonic heartbeat on Day 25; n = 18), nonpregnant (interestrus interval 15 to 21 d, n = 18), or having an apparent early embryonic loss (interestrus interval >25 d, n = 4). There was a group by time interaction (P < 0.001) for luteal blood flow from Days 3 to 18; it was approximately 1.10 ± 0.08 cm² (mean ± SEM) on Day 3, and increased to approximately 2.00 ± 0.08 cm² on Day 13 (similar among groups). Thereafter, luteal blood flow was numerically (albeit not significantly) greater in pregnant cows, remained constant in those with apparent embryonic loss, and declined (not significantly) between Days 15 and 18 in nonpregnant cows. Luteal blood flow was greater in pregnant than in nonpregnant (P < 0.05) and nonbred cows (P < 0.05, n = 14) on Day 15 (2.50 ± 0.16, 2.01 ± 0.16, and 2.00 ± 0.18 cm², respectively) and on Day 18 (2.40 ± 0.19, 1.45 ± 0.19, and 0.95 ± 0.21 cm²). In cows with apparent early embryonic loss, luteal blood flow was 2.00 ± 0.34 and 2.05 ± 0.39 cm² on Days 15 and 18, which was less (not significantly) than in pregnant cows, but greater (P < 0.05) than in nonbred cows on Day 18. Although mean luteal blood flow was significantly greater in pregnant than nonpregnant (and nonbred) cows on Days 15 and 18, due to substantial variation among cows, it was not an appropriate diagnostic tool for pregnancy status.     

Evidence for maternal regulation of early conceptus growth and development in beef cattle

Fifty-one cyclic beef cows were mated with fertile bulls. At 36 h after the start of oestrus, cows were assigned to receive sesame oil (controls) or progesterone (100 mg) on Days 1, 2, 3 and 4 of pregnancy. Peripheral plasma concentration of progesterone was measured until slaughter on Days 5 or 14. Cows were randomly assigned to be slaughtered on Days 5 or 14 or remain intact and palpated per rectum on Day 40 to verify pregnancy. Uteri on Days 5 and 14 were flushed for recovery of luminal protein and conceptus tissue. Conceptus and endometrial tissues were cultured with [3H]leucine and submitted to two-dimensional-PAGE and fluorography. Administration of progesterone increased peripheral plasma progesterone concentration on Day 2-5. Conceptuses recovered from progesterone-treated cows on Day 14 were advanced in development compared to conceptuses from control cows. Conceptuses recovered from progesterone-treated cows were viable as polypeptides associated with maintenance of pregnancy in cattle were synthesized and released at an earlier time and pregnancy was maintained beyond Day 40. Early progesterone stimulation altered the synthesis and release of polypeptides from endometrial explant cultures on Day 5. Results indicate a role of progesterone in the maternal regulation of conceptus growth and development in early pregnancy of cattle.     

Variations in oxytocin, vasopressin and neurophysin concentrations in the bovine ovary during the oestrous cycle and pregnancy

Bovine ovaries were obtained from the abattoir and corpora lutea were classified as: (1) early luteal phase (approximately Days 1-4); (2) mid-luteal phase (Days 5-10); (3) late luteal phase (Days 11-17); (4) regressing (Days 18-20) and (5) pregnant (Days 90-230). In addition, preovulatory follicles and whole ovaries without luteal tissue were collected. Concentrations of oxytocin, vasopressin, bovine neurophysin I and progesterone were measured in each corpus luteum by radioimmunoassay. Progesterone and neurophysin I levels increased from Stage 1 to Stage 2, plateaued during Stage 3 and declined by Stage 4. Oxytocin and vasopressin concentrations increased from Stage 1 to Stage 2 but declined during Stage 3 and were low (oxytocin) or undetectable (vasopressin) in follicles, whole ovaries and pregnancy corpora lutea. Therefore the concentrations of both peptide hormones were maximal during the first half of the cycle and declined before those of progesterone. The high concentration of oxytocin within the corpus luteum coupled with the presence of bovine neurophysin I suggests that oxytocin is synthesized locally.