Effects of indomethacin, luteinizing hormone (LH), prostaglandin E2 (PGE2), trilostane, mifepristone, ethamoxytriphetol (MER-25) on secretion of prostaglandin E (PGE), prostaglandin F2alpha (PGF2alpha) and progesterone by ovine corpora lutea of pregnancy or the estrous cycle

Two experiments were conducted to determine the luteotropin of pregnancy in sheep and to examine autocrine and paracrine roles of progesterone and estradiol-17 beta on progesterone secretion by the ovine corpus luteum (CL). Secretion of progesterone per unit mass by day-8 or day-11 CL of the estrous cycle was similar to day-90 CL of pregnancy (P > or = 0.05). In experiment 1, secretion of progesterone in vitro by slices of CL from ewes on day-8 of the estrous cycle was increased (P < or = 0.05) by LH or PGE2. Secretion of progesterone in vitro by CL slices from day-90 pregnant ewes was not affected by LH (P > or = 0.05) while PGE2 increased (P < or = 0.05) secretion of progesterone. Day 8 ovine CL of the estrous cycle did not secrete (P > or = 0.05) detectable quantities of PGF2alpha or PGE while day-90 ovine CL of pregnancy secreted PGE (P < or = 0.05) but not PGF2alpha. Secretion of progesterone and PGE in vitro by day-90 CL of pregnancy was decreased (P < or = 0.05) by indomethacin. The addition of PGE2, but not LH, in combination with indomethacin overcame the decreases in progesterone by indomethacin (P < or = 0.05). In experiment 2, secretion of progesterone in vitro by day-11 CL of the estrous cycle was increased at 4-h (P < or = 0.05) in the absence of treatments. Both day-11 CL of the estrous cycle and day-90 CL of pregnancy secreted detectable quantities of PGE and PGF2alpha (P < or = 0.05). In experiment 1, PGF2alpha secretion by day-8 CL of the estrous cycle and day-90 ovine CL of pregnancy was undetectable, but was detectable in experiment 2 by day-90 CL. Day 90 ovine CL of pregnancy also secreted more PGE than day-11 CL of the estrous cycle (P < or = 0.05), whereas day-8 CL of the estrous cycle did not secrete detectable quantities of PGE (P > or = 0.05). Trilostane, mifepristone, or MER-25 did not affect secretion of progesterone, PGE, or PGF2alpha by day- 11 CL of the estrous cycle or day-90 CL of pregnancy (P > or = 0.05). It is concluded that PGE2, not LH, is the luteotropin at day-90 of pregnancy in sheep and that progesterone does not modify the response to luteotropins. Thus, we found no evidence for an autocrine or paracrine role for progesterone or estradiol-17 36 on luteal secretion of progesterone, PGE or PGF2alpha.     

Effects of indomethacin, luteinizing hormone (LH), prostaglandin E(2) (PGE(2)), trilostane, mifepristone, ethamoxytriphetol (MER-25) on secretion of prostaglandin E (PGE), prostaglandin F(2alpha) (PGF(2alpha)) and progesterone by ovine corpora lutea of pregnancy or the estrous cycle

Two experiments were conducted to determine the luteotropin of pregnancy in sheep and to examine autocrine and paracrine roles of progesterone and estradiol-17 beta on progesterone secretion by the ovine corpus luteum (CL). Secretion of progesterone per unit mass by day-8 or day-11 CL of the estrous cycle was similar to day-90 CL of pregnancy (P >/= 0.05). In experiment 1, secretion of progesterone in vitro by slices of CL from ewes on day-8 of the estrous cycle was increased (P /= 0.05) while PGE(2) increased (P /= 0.05) detectable quantities of PGF(2alpha) or PGE while day-90 ovine CL of pregnancy secreted PGE (P /= 0.05). Trilostane, mifepristone, or MER-25 did not affect secretion of progesterone, PGE, or PGF(2alpha) by day-11 CL of the estrous cycle or day-90 CL of pregnancy (P >/= 0.05). It is concluded that PGE(2), not LH, is the luteotropin at day-90 of pregnancy in sheep and that progesterone does not modify the response to luteotropins. Thus, we found no evidence for an autocrine or paracrine role for progesterone or estradiol-17 36 on luteal secretion of progesterone, PGE or PGF(2alpha).     

Prolactin, progesterone and luteinizing hormone secretion after bromocriptine (CB-154) treatment in cyclic sows

The effect of bromocriptine (CB-154) on prolactin, progesterone and luteinizing hormone (LH) secretion was studied in cyclic sows. Four sows were given subcutaneous injections of bromocriptine on Day 14 of the estrous cycle (70 mg CB-154) and again on Day 16 of the cycle (50 mg CB-154). Two control sows were injected with vehicle at similar time intervals. Blood samples were taken four times daily (0700, 1100, 1500 and 1900 h) from Day 11 of the estrous cycle to Day 2 of the following estrous cycle. Prolactin peaks during the estrous cycle were not observed after CB-154 treatment. CB-154 treatment had no effect on plasma LH concentration, but plasma progesterone concentrations appeared to fluctuate more and slowly decreased.     

Prostaglandin metabolism in the ovine corpus luteum: catabolism of prostaglandin F(2alpha) (PGF(2alpha)) coincides with resistance of the corpus luteum to PGF(2alpha)

To examine possible mechanisms involved in resistance of the ovine corpus luteum to the luteolytic activity of prostaglandin (PG)F(2alpha), the enzymatic activity of 15-hydroxyprostaglandin dehydrogenase (PGDH) and the quantity of mRNA encoding PGDH and cyclooxygenase (COX-2) were determined in ovine corpora lutea on Days 4 and 13 of the estrous cycle and Day 13 of pregnancy. The corpus luteum is resistant to the action of PGF(2alpha) on Days 4 of the estrous cycle and 13 of pregnancy while on Day 13 of the estrous cycle the corpus luteum is sensitive to the actions PGF(2alpha). Enzymatic activity of PGDH, measured by rate of conversion of PGF(2alpha) to PGFM, was greater in corpora lutea on Day 4 of the estrous cycle (P < 0.05) and Day 13 of pregnancy (P < 0.05) than on Day 13 of the estrous cycle. Levels of mRNA encoding PGDH were also greater in corpora lutea on Day 4 of the estrous cycle (P < 0. 01) and Day 13 of pregnancy (P < 0.01) than on Day 13 of the estrous cycle. Thus, during the early estrous cycle and early pregnancy, the corpus luteum has a greater capacity to catabolize PGF, which may play a role in the resistance of the corpus luteum to the actions of this hormone. Levels of mRNA encoding COX-2 were undetectable in corpora lutea collected on Day 13 of the estrous cycle but were 11 +/- 4 and 44 +/- 28 amol/microgram poly(A)(+) RNA in corpora lutea collected on Day 4 of the estrous cycle and Day 13 of pregnancy, respectively. These data suggest that there is a greater capacity to synthesize PGF(2alpha), early in the estrous cycle and early in pregnancy than on Day 13 of the estrous cycle. In conclusion, enzymatic activity of PGDH may play an important role in the mechanism involved in luteal resistance to the luteolytic effects of PGF(2alpha).     

Effect of epostane, a competitive inhibitor of the 3beta-hydroxysteroid dehydrogenase enzyme, in cyclic sows

The effect of epostane, a 3beta-hydroxysteroid dehydrogenase (3beta-HSD) inhibitor, on the levels of plasma cortisol and progesterone and on the length of the estrous cycle in sows was examined. Epostane was administered orally on Days 0 to 2 (n = 3), Days 4 to 6 (n = 3), Days 10 to 12 (n = 2) or Days 17 to 19 (n = 3) of the estrous cycle, or subcutaneously on Day 0 (n = 3), Day 4 (n = 3), Day 10 (n = 4) or Day 17 (n = 3). Eleven days after the first dose of epostane, the treatments were repeated. One group of sows (n = 3) that was bled during a single estrous cycle served as controls. Cortisol levels in each of the eight groups of sows that received epostane did not differ (P>0.05) from those in control sows. In contrast, progesterone was lowered (P<0.01) when epostane was given by injection on Day 4, 10 or 17, or when given orally on Days 4 to 6 and 10 to 12. Although epostane reduced progesterone levels, the estrous cycle was not shortened. The interestrous interval for the sows (n = 14) that completed their experimental estrous cycle before they were sacrificed at approximately one week after the last dose of epostane was 21.6 +/- 2.71 d. It was concluded that epostane, as administered in this study, lowered progesterone levels but did not shorten the estrous cycle.     

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.     

Presence of the acute phase protein, bikunin, in the endometrium of gilts during estrous cycle and early pregnancy

Noninvasive, epitheliochorial placental attachment in the pig is regulated through endometrial production of protease inhibitors. The objective of the present study was to determine if the light-chain serine protease inhibitor of the inter-alpha-trypsin inhibitor family, bikunin, is produced by the porcine endometrium during the estrous cycle and early pregnancy. Western blot analysis revealed the presence of bikunin in uterine flushings of gilts collected during the luteal phase of the estrous cycle and early pregnancy (Days 12-18). However, bikunin unbound to the inter-alpha-trypsin heavy chains was detected only in endometrial explant culture medium obtained from estrus and pregnant (Days 12, 15, and 18) gilts. Endometrial bikunin gene expression was lowest on Day 10 of the estrous cycle and pregnancy, followed by a 30- to 77-fold increase on Day 15 of the estrous cycle and pregnancy. Bikunin gene expression decreased on Day 18 of the estrous cycle, whereas endometrial bikunin gene expression continued to increase in pregnant gilts. Bikunin mRNA was localized to the uterine glands between Days 15 and 18 of the estrous cycle and pregnancy. In addition to its role as a protease inhibitor, bikunin functions in stabilization of the extracellular matrix, which suggests that bikunin could be involved with facilitating placental attachment to the uterine epithelial surface in the pig.     

Repeatability of blood serum progesterone levels in dairy heifers on day 7 of the estrous cycle

Thirty normally cycling dairy heifers were used to determine the repeatability of blood serum progesterone levels on Day 7 ± 0.25 d of the estrous cycle. The experimental group consisted of 16 Holsteins and 14 dairy crossbreds ranging in age from 18 to 24 months. Day of the estrous cycle was determined from twice daily observations for standing heat (Day 0). Serum progesterone levels for Day 7 ± 0.25 d were determined by radioimmunoassay from blood samples collected by jugular venipuncture over three to four consecutive estrous cycles. Levels of blood serum progesterone for Day 7 ± 0.25 d ranged from 0.57 to 6.03 ng/ml. Least square means for the Holstein (2.74 ng/ml) and dairy crossbred (3.38 ng/ml) groups were different (P<0.006). The repeatability for levels of blood serum progesterone on Day 7 of the estrous cycle was low (0.0115).     

Monitoring ovarian cyclicity in postpartum Murrah buffalo through milk progesterone enzyme immunoassay

Milk samples were collected from Murrah buffalo between Day 30 and Day 120 post partum and analysed for progesterone concentration to monitor ovarian cyclicity. Progesterone levels were low (1 to 5 ng/ml) during the anestrous period. Levels were also low around estrus, but they began to increase at Day 6 postestrus; high levels (15 to 32 ng/ml) were maintained for different periods. There was a marked drop in progesterone level after Day 16 to 18 of the estrous cycle in those animals which returned to estrus. Progesterone levels remained high in buffalo which did not return to estrus, indicating that these animals were pregnant. Some of the progesterone cycles were not associated with the expression of estrus. This study indicated that a milk progesterone enzymoimmunoassay can be used to detect early pregnancy as well as conditions such as silent estrus and anovulatory estrus; it can thus help reduce the long intercalving period in buffalo.     

Quantitative changes in ovarian 17 alpha-hydroxylase/C17,20-lyase and aromatase activities during the estrous cycle of the hamster

Aromatase activity of the microsomal fraction of ovarian homogenates, measured by a tritium exchange assay using androstenedione (A-dione) as substrate, did not change during the 4-day estrous cycle of the hamster. In contrast, 17 alpha-hydroxylase, measured by a tritium exchange assay with progesterone as substrate, and particularly C17,20-lyase activity, evaluated by acetic acid production from progesterone, drastically decreased on the afternoon of proestrus (Day 4). The latter two activities remained low on Days 1 and 2 but increased dramatically on Day 3 and the morning of Day 4. The serum concentration of A-dione and 17-hydroxyprogesterone reached a peak at 1600 hr of proestrus and then decreased rapidly as the enzyme activities decreased. A-dione levels were undetectable on Day 1 but 17-hydroxyprogesterone levels remained elevated through Day 2. The results are consistent with the view that 17 alpha-hydroxylase and C17,20-lyase are activities of a single cytochrome P-450, as has been shown for testis and adrenal. Increases in hydroxylase and lyase activities occur concomitantly with decreases in the serum concentrations of progesterone, suggesting that the latter steroid may play a role in controlling these enzymes.     

Changes in tissue histamine during the estrous cycle, pregnancy and pseudopregnancy in the golden hamster

The histamine content of reproductive tissues and skeletal muscle was determined in the golden hamster during the estrous cycle, pregnancy, and pseudopregnancy. Histidine decarboxylase activity was measured in uterine implantation sites and intersites from Day 4 to Day 10 of pregnancy. Histidine decarboxylase was also measured in mesometria and placentas on selected days of gestation. During the estrous cycle, uterine and skeletal muscle histamine levels were highest on Day 2 and lowest on Day 4 of the cycle. The ovarian histamine content did not change significantly among the different stages of the cycle. While the histamine content of uterine implantation sites of attachment was high on Days 4 and measurable on Days 5 and 6 of pregnancy, the levels were below the limits of detection by Day 7. On the other hand, the highest levels of histamine were in the uterine interimplantation sites on Days 8 and 9. The ovarian levels of histamine were highest on Day 13 of pregnancy. Histamine in skeletal muscle did not change significantly during pregnancy. The histidine decarboxylase activity in the implantation sites began rising on Day 9 and increased dramatically on Day 10. Placental histidine decarboxylase activity was very high on Days 13 and 15. Overall, we observed changes in uterine and skeletal muscle histamine during the estrous cycle that may be explainable in light of previously reported changes in mast cell numbers and circulating estrogens. During pregnancy, histamine levels of implantation sites and implantation intersites varied, as did the histamine content of ovarian tissue. Histidine decarboxylase activity rises in the uterus and placental tissue after the formation of the placenta.     

Pattern of ovarian progesterone secretion during the luteal phase of the ovine estrous cycle

Pulsatile secretion of progesterone has been observed during the late luteal phase of the menstrual cycle in the rhesus monkey and human. As the luteal phase progresses in each of these species, there is a pattern of decreased frequency and increased amplitude of progesterone pulses. The present study was designed to determine the pattern of progesterone secretion during the late luteal phase (Days 10-16) of the normal ovine estrous cycle. Five unanesthetized ewes, each bearing an indwelling cannula in the utero-ovarian vein, were bled every 15 min from 0800 h on Day 10 through 0800 h on Day 16 of the estrous cycle. With the computer program PULSAR, it was determined that progesterone secretion was episodic, with pulsations observed on all days. Analysis of variance was used to determine differences in frequency, amplitude, and interpeak interval (IPI) of progesterone pulses among ewes and days. The ewes averaged 8.0 +/- 0.63 pulses of progesterone per 24 h. Mean frequency of pulses was not different between days but showed differences between ewes. Mean amplitude of progesterone pulses was 7.0 +/- 0.27 ng/ml, with no differences observed either between days or between ewes. Mean IPI was 197 +/- 7.1 min, and, like frequency, the IPI was not different between days, but varied between ewes. No consistent temporal relationship was found between progesterone pulses and luteinizing hormone (LH), as determined by bioassay and radioimmunoassay, on Day 14 of the cycle in one ewe. The results indicate that progesterone secretion is episodic during the luteal phase of the ovine estrous cycle and is independent of LH pulses.(ABSTRACT TRUNCATED AT 250 WORDS)     

Estrus and pregnancy after synchrony with lutalyse in conjunction with Syncro-Mate-B.

Estrous response and pregnancy rates are decreased for cows given Syncro-Mate-B (SMB) during metestrus (Day 1 to 5 of an estrous cycle). Data indicate these decreases are due, in part, to retention of a functional corpus luteum (CL). Our objective was to determine whether PGF2alpha administered in conjunction with SMB would improve estrous response and pregnancy rates in metestrous cows with no detrimental effects to cows in other stages of the estrous cycle. Three hundred seventy-three suckled beef cows were observed for estrus for 21 d before SMB administration to determine stage of an estrous cycle. Blood samples were collected 14 and 7 d before treatment and at SMB administration. Serum was assayed for concentration of progesterone to verify stage of estrous cycle or noncyclicity. All cows received the standard SMB regime and were allotted by age and stage of cycle to one of two groups. Cows denoted SMB + L received 25 mg of PGF2alpha 8 d after implantation, whereas cows denoted SMB served as controls. On Day 10, SMB implants were removed and females were observed for subsequent estrus. At this time, calves were removed from their dams for 48 h. Artificial insemination was performed 12 hr after observation of a standing estrus. Timed insemination was performed at 48 hr after implant removal for cows not inseminated at 24 or 36 hr after implant removal. Interval to synchronized estrus (within 5 d of implant removal) was lengthened for metestrous cows compared to cows in other stages of the cycle irrespective of treatment (P < 0.001). Cows receiving PGF2alpha had a greater pregnancy rate at 5 d compared to controls (P = .0672). Interval to estrus, estrous response, and pregnancy rate to A1 at d 28 or end of breeding season were not affected by administration of PGF2alpha in conjunction with SMB when compared to the standard SMB protocol.     

Pregnancy rates and metabolic profiles in cattle treated with propylene glycol prior to embryo transfer

The objective of this study was to determine the effect of a sustained propylene glycol administration to recipients of frozen/thawed in vivo derived bovine embryos. Heifers were treated with oral propylene glycol for the last 20 days before embryo transfer (n = 142), and untreated as controls (n = 133). Progesterone, insulin, insulin-like growth factor-I, glucose, urea and triglyceride were analysed in blood on Day 0 and Day 7 of the estrous cycle corresponding to embryo transfer. The heifers were selected as recipients when showing progesterone levels <2.0 ng/ml (Day 0) and >2.5 ng/ml (Day 7), according to corpus luteum quality on Day 7 by technicians unaware of animals treated. Within treated animals, significantly more recipients were selected, and increased progesterone, corpus luteum quality, pregnancy and calving rates were recorded. Day 7 progesterone concentrations were higher in heifers treated and transferred. Propylene glycol increased insulin and insulin-like-growth factor-I, but glucose, urea and triglyceride did not vary. Furthermore, insulin-like-growth factor-I, glucose and triglyceride increased at estrous time, but urea decreased and insulin remained unaltered. Together with the sustained gain in pregnancy rates throughout the experiment (2 years), other evidences suggested that the observed effects did not rely on nutritional deficiency. Thus, propylene glycol improved pregnancy rates after embryo-transfer, and progesterone, insulin and insulin-like-growth factor-I are probably involved in this effect.     

Effect of prostaglandin F2 alpha at different stages of guinea-pig pregnancy

It is believed that suppression of the processes by which prostaglandin F2 alpha is released from the uterus during the estrous cycle is vital to maintenance of pregnancies in guinea-pigs. Prostaglandin F2 alpha was injected into pregnant guinea-pig at four different stages of gestation to investigate the effect increased prostaglandin might have. The study revealed an alteration in the sensitivity of the pregnancy to prostaglandin F2 alpha as pregnancy progressed. Recovery from the prostaglandin insult was more likely if the injection was given after Day 24 than before Day 18. In some animals the serum progesterone levels fell following the injection and then subsequently recovered. It appears that effects which are potentially hazardous to the pregnancy are countered in a variety of ways.     

Characterization of endometrial receptors for ovine trophoblast protein-1 during the estrous cycle and early pregnancy in sheep

Scatchard analysis was used to determine the distribution, number, and affinity of unoccupied receptors for ovine trophoblast protein-1 (oTP-1) in endometrium of sheep throughout the estrous cycle and early pregnancy. In Experiment I, oTP-1 receptor characteristics were determined in membrane preparations of caruncular and intercaruncular regions of endometrium collected from uterine horns ipsilateral and contralateral to the ovary bearing the corpus luteum. Receptor concentrations and affinity constants for oTP-1 were not different (p greater than 0.1) between the four endometrial regions examined, suggesting that the expression of receptors for oTP-1 occurs uniformly throughout the endometrium. Endometrial receptor characteristics for oTP-1, luteal wet weights, and progesterone contents were determined throughout the estrous cycle and early pregnancy in Experiment II. Concentration of receptors and affinity constants for oTP-1 varied throughout the estrous cycle and early pregnancy (p less than 0.01), with the pattern of change differing between cyclic and pregnant ewes (p less than 0.01). Numbers of receptors for oTP-1 were maximal on Day 4 of the estrous cycle and declined progressively to Day 12 (p less than 0.05) in both cyclic and pregnant ewes. After Day 12, the quantity of unoccupied receptors for oTP-1 increased (p less than 0.05) gradually to Day 16 in cyclic ewes, but declined (p less than 0.05) further in the endometrium of pregnant ewes. The affinity constants of endometrial receptors for oTP-1 were similar in cyclic and pregnant ewes prior to Day 12, increasing threefold from Days 4 to 12 (p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Peripheral inhibin levels in relation to climatic variations and stage of estrous cycle in buffalo (Bubalus bubalis )

The present study investigated the peripheral plasma inhibin levels in relation to 1) the stage of estrous cycle and the effect of climatic variations. Blood samples were collected from cyclic buffalo (n=5) once daily for 32 consecutive days during the tropical hot humid (summer) and cold (winter) seasons. Estrus was recorded by parading a vasectomized bull as well as by plasma progesterone determination. In the winter season, peripheral inhibin concentrations which were lowest (0.35 +/- 0.02 ng/ml) during the mid-luteal phase of estrous cycle (Day 6 to Day 14, Day 0 = day of estrus) increased significantly (P < 0.02) to 0.47 +/- 0.04 ng/ml during the late luteal phase (Day -4 to Day -2) and then further to 0.52 +/- 0.03 ng/ml (P< 0.02) during the periestrus phase (Day -1 to Day 1). Inhibin concentrations then decreased significantly (P < 0.02) to 0.40 +/- 0.03 ng/ml during the early luteal phase (Day 2 to Day 5). In the summer season the differences in peripheral inhibin concentrations among different phases of estrous cycle were found to be nonsignificant. A comparison of the circulating inhibin concentrations between the two seasons indicated that inhibin concentrations were significantly higher in the late luteal phase (P < 0.01) and periestrus phase (P < 0.05) during the winter season compared with corresponding periods during the summer season. The present study suggests that peripheral inhibin concentrations change in the estrous cycle during cooler breeding season and that environmental heat stress can cause a reduction in peripheral inhibin concentrations.     

Control of luteal relaxin release by prostaglandin F2 alpha: differences in the sow cycle and pregnancy

The effect of an in vivo prostaglandin F2 alpha (PGF2 alpha) challenge in pregnant and cyclic sows was compared to determine whether PGF2 alpha-induced release of relaxin (RLX) from the corpus luteum (CL) in late pregnancy is also effective during the cycle. Ovarian venous RLX and progesterone were monitored by radioimmunoassay and RLX localized in the CL by immunohistochemistry. In Day 108 pregnant sows, infusion of PGF2 alpha (100 micrograms) into the ovarian artery resulted in an immediate and sustained rise in ovarian venous RLX with an initial decline in progesterone levels by 30 min which then returned to pretreatment levels. In Day 13 or 15 cyclic sows with functional corpora lutea (i.e., elevated progesterone), RLX was undetectable in ovarian venous blood after 100 micrograms of PGF2 alpha. Administration of PGF2 alpha via either the jugular vein or intramuscular route was also ineffective in releasing RLX from the CL of the cycle. The intensity of RLX immunostaining of the CL was similar in saline and PGF2 alpha-treated sows. These studies indicate that the control of RLX release from the sow CL differs in the estrous cycle and pregnancy.