Failure of exogenous LH to prevent PGF2α-induced luteolysis in beef cows

Henderson and McNatty (Prostaglandins 9:779, 1975) proposed that LH from the preovulatory LH surge attached to receptors on luteal cells and that this attachment might protect the early corpus luteum from PGF_2α induced luteolysis. To test this hypothesis, experiments were performed on heifers at day 10–12 of the cycle. Both jugular veins were catheterized and infusions of either saline (0.64 ml/min) or LH-NIH-B9 (10 μg/min; 0.64 ml/min) were given. Saline infusions were from 0–12 h; LH infusions were for 10 h and were preceded by a 2 h saline infusion. All animals were given 25 mg PGF_2α im at 6 h (6 h into the saline infusion and 4 h into the LH infusion). Blood samples were taken at 0.5 h, 1 h and 4 h intervals from 0–12, 13–18 h and 22–24 h respectively. Serum was assayed for LH and progesterone by radioimmunoassay methods. Two animals received saline and two received LH in each experiment. Eact treatment was replicated 6 times. LH infusion resulted in a mean serum LH of 57 ng/ml compared to 0.90 ng/ml in saline infused animals. This elevation of LH did not alter PGF_2α induced luteolysis as indicated by decline in serum progesterone. This experiment does not support the hypothesis that the newly formed corpus luteum is resistant to PGF_2α because of protection afforded by the protestrus LH surge.     

Effects of PGE1 or PGE2 on luteal function in pseudopregnant rats

Effects of PGE₁ or PGE₂ on luteal function were studied in 163 pseudopregnant rats. PGE₁ (10, 100, or 300μg) given intrauterine every 6 hr did not shorten pseudopregnancy (P < 0.05), however, the same doses of PGE₂ given intrauterine every 6 hr advanced luteolysis (P < 0.05). PGE₁ (100 or 300μg) given every 4 hr intramuscular maintained levels of progesterone in peripheral blood above controls (P < 0.05) while 100 or 300μg of PGE₂ hastened the decline in progesterone (P < 0.05). The antiluteolytic effect of PGE₁ was not via an inhibition of PGF secretion (P < 0.05) by the uterus or by induction of ovulation in treated animals. Moreover, PGE₁ (100, 200, or 500μg) given intramuscular every 4 hr from day 4 of pseudopregnancy until the next proestrus delayed luteal regression around 3 days (P < 0.05). PGE₂ at doses of 100, 200, or 500μg every 4 hr given intramuscular consistently shortened pseudopregnancy (P < 0.05). Lower doses were without effect (P < 0.05). Based on the above data it is concluded that PGE₂ is consistently luteolytic whereas PGE₁ is not luteolytic in pseudopregnant rats and that PGE₁ may be an antiluteolysin.     

Effects of prostaglandins E1 and F2α on serum luteinizing hormone concentration and on some sexual functions in male rabbits

PGE₁(50μg/animal) and PGF_2α (250 μg/animal) caused a transient in serum LH at 5 min after injection. PGE₁ (250 μg/animal) had a biphasic effect on serum LH. A small peak was obtained at 5 min, and a second, larger peak at 60 min after injection. It is suggested that the first peak is a result of the stress associated with injection of the PGs, whereas the second peak represents a physiological effect of PGE. Subcutaneous injection of PGE₁ (1 mg in arachis oil b.i.d.) for 10 days did not affect the concentration of LH in serum, the function of the accessory sexual glands or the sexual activity. PGF_2α, given at the same dose and in the same manner, increased the sexual activity but left all other variables unaffected. The pituitary responsiveness to LH-RH was unaltered by the treatment with PGE₁ and PGF_2α.     

Effect of indomethacin and 7-oxa-13-prostynoic acid on luteinization of transplanted rat ovarian follicles induced by luteinizing hormone and prostaglandin E2

The ability of prostaglandin E₂ (PGE₂) to initiate luteinization was demonstrated using a system of in vitro incubation of ovarian follicles followed by transplantation. Follicles from diestrous rats were incubated with 0.05 to 50 μg/ml PGE₂, 10 μg/ml luteinizing hormone (LH), or alone in Krebs-Ringer bicarbonate buffer plus glucose for 2 hr. Then follicles were transplanted under the kidney capsules of hypophysectomized recipients, with follicles exposed to PGE₂ on one side and those exposed to LH or buffer only on the other side. As determined at autopsy 4 days later and confirmed by histological examination, follicles exposed to PGE₂ at concentrations of 0.5 μg/ml or greater, or to LH, transformed into corpora lutea, but control follicles regressed. Incubation of follicles with LH in the presence of indomethacin, an inhibitor of prostaglandin synthesis, significantly reduced the incidence of luteinization. Prostaglandin E₂ (10 μg/ml) was able to override the inhibition of luteinization by indomethacin (150 μg/ml). The prostaglandin analogue 7-oxa-13-prostynoic acid (100 μg/ml) failed to prevent luteinization in response to either 5 μg/ml LH or 1 μg/ml PGE₂. Results with PGE₂ and with indomethacin suggest a role for prostaglandins in the luteinizing action of LH.We have reported previously that in vitro exposure of diestrous rat follicles to luteinizing hormone (LH) will result in transformation of the follicles to corpora lutea following transplantation under the kidney capsules of hypophysectomized rats. Dibutyryl cyclic AMP (DBC) mimics this effect of LH, and transplants produce progesterone in measurable amounts after both LH and DBC exposure when prolactin is administered in vivo to recipients.Kuehl et al. have suggested that prostaglandins may act as obligatory intermediates in the effect of LH on the ovary, acting between LH and adenylate cyclase. Preliminary results indicated that prostaglandin E₂ (PGE₂) could induce luteinization in our system. The extent of prostaglandin involvement in luteinization was further investigated in this work, using two reported antagonists of prostaglandin action, indomethacin and 7-oxa-13-prostynoic acid. Indomethacin has been found to inhibit synthesis of prostaglandins E₂ and F_2α; 7-oxa-13-prostynoic acid, which acts as a competitive antagonist of prostaglandins, prevented the effect of LH and prostaglandins E₁ and E₂ on cyclic AMP production in mouse ovaries.     

Induction of 20α-hydroxysteroid dehydrogenase in rat corpora lutea of pregnancy by prostaglandin F2α

20α-OH-SDH is a marker of luteolysis in rat corpora lutea and appearance of this enzyme is inhibited by prolactin but stimulated by LH or hCG. PGF₂α induced 20 α-OH-SDH activity in corpora lutea of pregnant rats and a significant fall in peripheral plasma progesterone concentrations when administered i.m. for two consecutive days. Rats treated with PGF₂ α on days 8 and 9 of pregnancy were resorbing implants by day 10. Exogenous progesterone, but not estrogen, prevented implant resorption, yet 20 α-OH-SDH appeared in the corpora marking luteolysis. HCG, LH and prolactin, but not FSH, prevented pregnancy termination and inhibited induction of 20 α-OH-SDH in rats treated with PGF₂ α in early pregnancy. PGF₂α also induced 20α-OH-SDH in luteal tissue of intact and hypophysectomized rats treated on days 14 and 15 of pregnancy, but neither exogenous steroids or gonadotrophins blocked the induction of the enzyme in rats treated at this time. The increase in lutein 20α-OH-SDH activity during the peripartal period was partially blocked by administration of the prostaglandin biosynthesis inhibitor, indomethacin, suggesting a role for endogenous prostaglandins in the induction of 20α-OH-SDH at term. It appears that PGF₂α acts directly on the ovary to induce 20α-OH-SDH activity by preventing the luteotrophic action of prolactin. Other luteal NADPH-dependent dehydrogenase activities are not markedly stimulated following PGF₂α administration.     

Prostacyclin (PGI2) effets on anterior pituitary hormones in the rat in vivo

Intravenous injection of 600 μg PGE₂ or PGI₂ significantly increased serum LH and prolactin levels in estradiol treated ovariectomized rats. There was no effect on serum FSH concentration. PGE₂ and PGI₂ stimulated LH release in a non-dose dependent manner, while prolactin levels were positively correlated with the dose administered following PGI₂ treatment. 6-keto-PGF_1α at a comparable dose had no effect on pituitary hormone levels. Subcutaneous administration of 1 mg/kg or 60 mg/kg PGI₂ for seven days significantly depressed serum LH level both in male and female rats. These doses had no effect on serum FSH or prolactin levels.     

Midterm abortion in hamsters induced by Silastic-PVP-PGE2 tubes

Intrauterine insertion of a 0.5 cm long Silastic-PVP tube containing 750 μg PGE₂ (lyophilized sodium salt) caused midterm abortion in hamsters within 48 hours. An earlier study using a similar Silastic-PVP tube delivery system showed that 200 μg of PGF_2α (Tham) was sufficient to induce abortion in 100% of pregnant hamsters (18). Prostaglandin E₂ is, therefore, about 3.5–4 times less potent than PGF_2α as an abortifacient in the hamster. The release of ³H-PGE₂ from Silastic-PVP tube and is also described. It is suggested that an increase in LH release might be one of the factors leading to luteolysis; and that either PGE₂ exerts a direct luteolytic effect or this effect is manifested after its being converted to PGF_2α.     

Effect of prostaglandin F2α on the hCG-stimulated progesterone production by human corpora lutea

The effect of prostaglandin PGF_2α on the hCG stimulated and basal progesterone production by human corpora lutea was examined . hCG (40 i.u./ml) stimulated progesterone formation in corpora lutea of early (days 16–19 of a normal 28 day cycle), mid (days 20–22) and late (days 23–27) luteal phases. This stimulation was inhibited by PGF_2α (10 μg/ml) in corpora lutea of mid and late luteal phases. PGF_2α alone did not show a consistent effect on basal progesterone production. The inhibition of hCG stimulated progesterone production by PGF_2α at times corresponding to luteolysis indicates a role for that prostaglandin in the process of luteolysis in the human corpus luteum.     

Effect of prostaglandin F2α on ovarian and pituitary function in the mid-luteal phase

The effects of PGF_2α infusion in a dose of 25 μg/min for 5 hours on serum levels of estradiol-17β, progesterone, LH, FSH, TSH and prolactin, and on the pituitary hormone responsiveness to LRH and TRH were studied in 10 apparently healthy cycling women in the mid-luteal phase. No systematic alteration was seen in the pituitary and ovarian hormone levels during PGF_2α infusion, and the pituitary hormone responses to releasing hormones were unaffected. Ovarian steroid production increased in response to increased gonadotropin levels after LRH injection during PGF_2α administration. These results confirm that PGF_2α is not luteolytic in humans and no apparent relationship between PGF_2α and pituitary hormone secretion exists.     

An intra-corpus luteum site for the luteolytic action of prostaglandin F2α in the rhesus monkey

It has not been possible to demonstrate prostaglandin F₂α (PGF₂α) participation in primate luteolysis under conditions of systemic administration or of acute intraluteal injection. These study designs were hampered by the short biological half-life in the first instance and brevity of administration in the latter. In this study, luteolysis has resulted from chronic, intraluteal delivery of PGF₂ α. Using the Alzet osmotic pump-cannula system, normally cycling rhesus monkeys were continuously infused, until menses occurred, with PGF₂ α (10 ng/1/hr) directly into the corpus luteum (CL, n=6), into the stroma of the ovary bot bearing the corpus luteum (NCL, n=3), or subcutaneously (SC, n=5). An additional 5 monkeys received vehicle (V) into the corpus luteum. All experiments commenced 5–7 days after the preovulatory estradiol surge. Luteal function was assessed by the daily measurements of plasma progesterone, estradiol, and LH. Intraluteal PGF₂α caused premature functional luteolysis in all monkeys, as reflected by a highly significant decline in circulating progesterone and estradiol and the early onset of menstruation, when compared to the other groups. V, NCL, and SC infusions had no effect on either circulating steroid levels or luteal phase lengths. None of the experimental groups showed any change in plasma LH concentrations. These are the first data to indicate that PGF₂α can induce functional luteolysis in the primate, and the site of action appears to be the corpus luteum.     

The action of prostaglandin E2 and F1α on myocardial ischaemia-infarction arrhythmias in the dog

Prostaglandin E₂ and F_1α infusions have been tested for their ability to reduce the arrhythmias associated with occlusion of the left descending coronary artery in the anaesthetised dog. At 1 μg/kg/min both PGs reduced the incidence of premature ventricular contractions occurring during 25-min occlusions, while not reducing the incidence of ventricular fibrillation occurring on occlusion release. When infused for 5-min periods at 1 to 16 μg/kg/min, neither PGE₂ nor PGF_1α effectively reduced the frequency of ventricular arrhythmias occurring 24 hr after a permanent coronary occlusion.     

Reversal of indomethacin-induced inhibition of tubuloglomerular feedback by prostaglandin infusion

Experiments were performed in rats to study the effect of infusion of PGI₂, PGE₂, and PGF_2α on tubuloglomerular feedback responses (i.e. the change of SNGFR in response to a change of loop of Henle flow rate) in the presence and absence of simultaneous inhibition of endogenous PG synthesis with indomethacin. Infusion of PGI₂ or PGE₂ at rates that did not alter arterial blood pressure did not significantly modify the magnitude of feedback responses (PGI₂) 8.5 μg/hr, PGE₂ 85 μg/hr). Some inhibition of feedback responses was seen when PGI₂ and PGE₂ were administered at higher rates were associated with a reduction of blood pressure (PGI₂ 20 μg/hr, PGE₂ 200 μg/hr). PGI₂ (8.5 μg/hr) and PGE₂ (85 μg/hr) largely prevented feedback inhibition induced by indomethacin. When given subsequent to indomethacin PGI₂ and PGE₂ restored feedback responsiveness almost to normal. In contrast, PGF_2α did not influence feedback inhibition caused by indomethacin. Infusion of PGI₂ induced partial restoration of feedback responses in DOCA-salt treated animals in which the feedback system is virtually completely inactive. Our results indicate that availability of PGI₂ or PGE₂ is necessary for the normal operation of the tubuloglomerular feedback mechanism for control of nephron filtration rate.     

The lack of involvement of central nervous system in the antiarrhythmic effects of prostaglandins E2, F2α, and I2 in cat

The role of the central nervous system (CNS) in the antiarrhythmic effects of prostaglandins (PGs) E₂, F_2α, and I₂ was studied by administering each agent into the left lateral cerebral ventricle (i.c.v. administration) of chloralose-anesthetized cats. The cardiac arrhythmias were produced by intravenous (i.v.) infusion of ouabain (1 μg/kg/min). The PGs E₂, F_2α and I₂ on i.c.v. administration in the dose range of 1 ng to 10 μg failed to inhibit ouabain-induced cardiac arrhythmias. However, when infused i.v., PGE₂ (1 μg/kg/min), PGF_2α (5 μg/kg/min), and PGI₂ (2 μg/kg/min) effectively suppressed these arrhythmias. The standard antiarrhythmic drug propanolol (0.5–8.0 mg)oni.c.v.administration also significantly reduced the ouabain-induced cardiac arrhythmias. It is suggested that the CNS is not the site of action of PGs E₂, F_2α, and I₂ in antagonising the ouabain-induced cardiotoxicity in cats.     

Follicle stimulating hormone and prolactin release induced by prostaglandins in rat

Ten to 60 minutes following a single i.v. injection of PGE₂ (500 μg/rat) into male rats of 30 to 35 days of age FSH concentration in the serum was raised significantly. The rise in FSH was maintained from 10 to 60 minutes after treatment, then at 90 minutes FSH had declined and was not significantly different from that of the control before treatment. Prostaglandin E₁, E₂ or F_2α (670μg/rat) significantly increased the serum prolactin level 10 to 60 minutes after a single i.v. injection in spayed rats primed with estrogen and progesterone. And, rats primed with estrogen and progesterone. And, increases in prolactin in the serum were observed with as little as 2μg of PGE₁ or E₂, and 20μg of PGF_2α. Twenty μg of PGE₂, and 200μg of PGE₁ or F_2α gave the maximum stimulation. These results indicate that release of pituitary hormones is affected by prostaglandins.Prostaglandins (PGs) are widely distributed in mammalian tissues, and they have been reported to have an almost equally wide variety of endocrine and metabolic effects. It was recently postulated that PGs may be involved in the process of ovulation because ovulation was blocked by inhibitors of PG synthesis (1–5).     

Effect of PGF2α on progesterone secretion and adenylate cyclase activity in ovine luteal tissue

Ovine luteal slices were used to study the effects of prostaglandins (PG) F₂α on luteinizing hormone (LH)-stimulated secretion of progesterone and adenylate cyclase activity. The accumulation of progesterone in incubation medium and adenylate cyclase activity was similar after incubation of luteal slices with Medium 199 alone or Medium 199 containing PGF₂α (250 ng/ml) for 3 hr. Addition of luteinizing hormone (LH; 100 ng/ml) resulted in a 2–3 fold increase in both the rate of progesterone accumulation and adenylate eyclase activity by 3 hr. When luteal slices were incubated in the presence of both LH and PGF₂α the rates of progesterone accumulation and adenylate cyclase activity were identical to those in flasks containing LH alone after 1 hr; however, after 3 hr both LH stimulated progesterone accumulation and adenylate cyclase activity were inhibited to levels similar to those observed in control slices.In a second experiment, after 60–120 min of exposure to PGF₂α the rate of progesterone accumulation in the medium was not different from that in untreated control slices. In addition, after this experiment the luteal slices were homogenized and the basal, sodium fluoride, LH, isoproterenol (ISO) and PGE₂ sensitive adenylate cyclase activities were determined to evaluate the hormonal specificity of the negative effect of the pretreatment with PGF₂α. Both LH and ISO stimulated adenylate cyclase activities were reduced after PGF₂α pretreatment. However, fluoride ion stimulated adenylate cyclase activity was not significantly effected by PGF₂α pretreatment and PGE₂ sensitive adenylate cyclase was effected only slightly.     

Increased blood LH and testosterone after administration of prostaglandin F2α in bulls

Two types of experiments were conducted to determine the relationship of changes in blood luteinizing hormone (LH) and testosterone in bulls given prostaglandin F_2α (PGF_2α). Episodic surges of LH and testosterone occurred in tandem, apparently at random intervals, on the average once during the 8-hr period after bulls were given saline. In contrast, after sc injection of 20 mg PGF_2α, blood serum testosterone increased synchronously to a peak within 90 minutes four-fold greater than pre-injection values, and the testosterone surges were prolonged about three-fold compared to those in controls. Each of the PGF_2α-induced surges of testosterone was preceded by a surge of blood serum LH which persisted for about 45 minutes and peaked at about 3 ng/ml. In a second experiment, PGF_2α was infused (iv, 0.2 mg/min) for 20 hr; blood plasma testosterone increased from 7.0 ± 0.6 to 16.0±1.5 ng/ml within 2.5 hr and remained near this peak for 10 hr. Then testosterone gradually declined to about 9 ng/ml at the conclusion of the 20-hr infusion. These changes in testosterone were paralleled by similar changes in blood plasma LH, although LH declined 3 hr earlier than testosterone. Random episodic peaks of blood plasma LH and testosterone typical of untreated bulls resumed within 8 hr after conclusion of PGF_2α infusion. In both experiments, the surge of testosterone after PGF_2α was preceded by increased blood LH. We conclude that increased LH after administration of PGF_2α probably caused the increased testosterone. However the mechanisms of these actions of PGF_2α remain to be determined.     

The effect of intra-uterine prostaglandin F2α on corpus luteum function in the human

Prostaglandin F_2α (PGF_2α) was administered via a Foley catheter over a 12 hour period to 8 healthy volunteers awaiting laparoscopic sterilisation. The amount of PGF_2α infused varied between 500 μg and 2000 μg every 2 hours for 6 doses. Plasma progestins and oestradiol 17β, and urinary estrogens and pregnanediol were assayed throughout the study period.There was no evidence of luteolysis in any patient although vaginal bleeding of varying duration occurred in all women within 36 hours of administration of PGF_2α.     

Effect of insulin-induced hypoglycemia on the serum concentrations of thyroxine,triiodothyronine and reverse triiodothyronine

The effect of insulin-induced hypoglycemia on serum thyroid hormone concentrations was studied in nine healthy individuals. Before, during and after the hypoglycemia blood samples were taken for measurement of the concentrations of glucose, thyroxine (T₄), triiodothyronine (T₃), reverse triiodothyronine (rT₃), catecholamines and pituitary hormones.There was no change in the mean serum T₄ level (± the standard error of the mean) of 67 ± 2 μg/l. However, the T₃ concentrations rose from a mean basal level of 1.86 ± 0.06 μg/l to a mean peak of 2.51 ± 0.21 μg/l (P < 0.01) at 45 minutes after the insulin injection, and the rT₃ concentrations fell from a mean basal level of 0.184 ± 0.008 μg/l to a mean nadir of 0.171 ± 0.022 μg/l (not a significant change). The mean peak epinephrine level was 545 ± 103 ng/l and it occurred between 30 and 45 minutes after the insulin injection; the mean peak norepinephrine level was 584 ± 114 ng/l and it occurred between 30 and 90 minutes after the injection. The growth hormone levels reached a mean peak of 26.1 ± 4.8 μg/l and the plasma cortisol levels rose to 215 ± 9 μg/l. The mean basal prolactin level was 8.5 ± 0.9 μg/l; in five subjects there was a rise to a mean peak of 50.6 ± 14.6 μg/l, whereas in the remaining four no significant increase occurred. No correlation was found between the changes in the serum T₃ concentration and any of the other factors studied.It was concluded that acute hypoglycemia is associated with a rapid increase in the serum T₃ concentration.     

Luteolysis, estrus and ovulation, and blood prostaglandin F after intramuscular administration of 15, 30 or 60 mg prostaglandin F2α

During diestrus in three consecutive estrous cycles, each of six heifers was given (im) 30 mg, 15 mg (twice at 6-hr intervals) and 60 mg prostaglandin F_2α (PGF_2α) tham salt. Neither the decline in blood progesterone, the increase in blood estradiol, the duration or the peak of the LH surge, the interval to onset of estrus, nor the interval to ovulation was affected significantly by dose of PGF_2α. Thus, relative to that after 30 mg PGF_2α im, two injections of 15 mg at 6-hr intervals or 60 mg PGF_2α did not hasten luteolysis. Thirty mg was an ample im dose of PGF_2α to cause luteolysis. Regardless of im dose of PGF_2α, blood PGF peaked at about 6.0 ng/ml within 10 minutes and returned to basal values (<1.0 ng/ml) within 90 minutes. In another trial, after a single iv injection of 5 mg PGF_2α, blood PGF peaked (25 ng/ml) within 5 minutes and returned to basal values within 15 minutes. During a 30-minute infusion (0.5 mg/minute) of PGF_2α, blood PGF plateaued at 29.5 ng/ml with a metabolic clearance rate of 17.0 liters per minute.     

Biosynthesis of prostaglandins and thromboxane B2 by fetal lung homogenates

The conversion of arachidonic acid to prostaglandins (PG's) and thromboxane B₂ (TXB₂) was investigated in homogenates from fetal and adult bovine and rabbit lungs. Adult bovine lungs were very active in converting arachidonic acid (100 μg/g tissue) to both PGE₂ (10.7 μg/g tissue) and TXB₂ (6.2 μ/g tissue). Smaller amounts of PGF_2α (0.9 μ/g) and 6-oxoPGF_1α were formed. Homogenates from fetal calf lungs during the third trimester of pregnancy were quite active in converting arachidonic acid to PGE₂, but formed very little TXB₂, PGF_2α or 6-oxoPGF_1α. Homogenates from rabbit lungs converted arachidonic acid (100 μg/g) mainly to PGE₂, both before and after birth. The amount of PGE₂ formed increased during gestation to a maximum of about 6 μg/g tissue at 28 days of gestation. It then decreased to a minimum (1.5 μg/g) which was observed 8 days after birth, followed by an increase to about 4 μg/g in older rabbits.