Reversal of the effects of 4-amino-antipyrine on myometrial and cervical electromyographic activity by PGF2 alpha in the ovariectomized estradiol-replaced non-pregnant ewe

Cervical and uterine electromyograms (EMG) have been recorded from ovariectomized non-pregnant ewes. When the animals were infused with saline, the frequency of EMG events lasting less than 180 seconds was not different from those lasting more than 180 seconds. During infusion of estradiol at 100 micrograms X 24 hours-1 into the maternal jugular, the frequency of events less than 180 seconds increased significantly in the myometrium and in the cervix. Contracture activity (events lasting more than 180 seconds) was not significantly different in the myometrium compared to the cervix before estradiol administration. During estradiol infusion, the contracture activity remained unchanged. During 4-amino-antipyrine (4AA) administration, the contracture activity decreased significantly in the myometrium, while an insignificant change occurred in the cervix. This state was associated with a decrease in the venous PGFM:6-keto F1 alpha plasma ratio. Infusion of PGF2 alpha (.5 micrograms min-1 and 1.0 microgram X min-1 for ten minutes) into the femoral artery resulted in a significant increase in the frequency of events less than 180 seconds in both the myometrium and cervix. For the duration of the ten-minute infusion, the activity was contracture-like. These findings suggest that the cervix may not only be influenced by mechanical properties (stretch) and local paracrine factors but also by various stimulators and inhibitors irrespective of the myometrium.     

Prostaglandin receptors EP and FP are regulated by estradiol and progesterone in the uterus of ovariectomized rats

Background  

Prostaglandins are important for female reproduction. Prostaglandin-E2 acts via four different receptor subtypes, EP1, EP2, EP3 and EP4 whereas prostaglandin-F2alpha acts through FP. The functions of prostaglandins depend on the expression of their receptors in different uterine cell types. Our aim was to investigate the expression of EPs and FP in rat uterus and to identify the regulation by estradiol, progesterone and estrogen receptor (ER) selective agonists.     

Early identification of non-pregnant and pregnant ewes in the field using circulating progesterone concentration

A method for early indentification of non-pregnant and pregnant ewes is described. It is applicable to field research situations where mating data for individual ewes cannot be collected and requires three plasma progesterone measurements from each ewe over a 12-days period.Ewes were diagnosed non-pregnant according to whether their lowest progesterone concentration (p) was below or above a “discriminatory value”. This value was chosen after examining the overall frequency distribution of values of log₁₀p.In experiment 1, $\text{25}\text{27}$ non-pregnant ewes and $\text{64}\text{65}$ pregnant ewes (20 to 31 days post-mating) were diagnosed correctly (96.7% accuracy). In experiment 2, $\text{40}\text{41}$ non-pregnant ews and $\text{45}\text{48}$ pregnant ewes (21 to 34 days post-mating) were diagnosed correctly (95.5% accuracy).     

Plasma progesterone concentrations derived from the administration of exogenous progesterone to ovariectomized mares.

Six ovariectomized mares were divided into 3 groups to determine the effects of exogenous progesterone in oil and repositol progesterone on plasma progesterone concentrations. Progesterone in oil was administered in 7 daily injections in Exp. I. Progesterone concentrations were not maintained greater than 1.0 ng/ml for 24 h with 50 mg/day. However, they remained greater than 1.0 ng/ml during the last 4 days of 100 mg/day and greater than 1.5 ng/ml throughout the injection sequence of 200 mg/day. Repositol progesterone was administered on Days 1 and 7 in Exp. II. At 500 mg, progesterone concentrations peaked in 6 h but returned to near 1.0 ng/ml in 2 days. At 1000 mg and 2000 mg, plasma progesterone was maintained at approximately 2.0 and 4.0 ng/ml respectively for 7 days after injection on Day 1 and was 1.5 and 3.5 ng/ml respectively, 11 days after injection on Day 7. An indication of a cumulative effect on plasma progesterone was observed following repeated dosages of both progesterone in oil and repositol progesterone.     

Peripheral progesterone levels in pregnant and non-pregnant heifers following use of HCG.

Progesterone concentration in jugular blood of bred beef heifers was determined on days 9 and 16 in two trials. Human chorionic gonadotrophin (HCG) was administered to some of the heifers in each trial in an attempt to improve pregnancy percentage.In Trial 1, 183 heifers were divided into a control group and three groups of animals which were subjected to a 9-day estrous synchronization treatment prior to breeding. The treatment consisted of an ear implant containing 17 α acetoxy-11-beta-methyl 17 nor preg 4-ene 3, 20 dione (norgestomet) left in place for 9 days and an injection of 5 mg estradiol valerate (EV) and 3 mg of norgestomet given at the time of implantation. The heifers in one group received .25 mg estradiol-17β at time of implant removal; heifers in the 2nd group received 1500 IU of HCG in 5% beeswax and 95% sesame oil at breeding time, while heifers in the 3rd group received a placebo injection containing 5% beeswax and 95% sesame oil at breeding time. No differences in serum levels of progesterone were observed (P>0.5) between treatments or between pregnant and non-pregnant heifers on day 9 or 16 (P>.05). Pregnancy percentage in heifers receiving HCG was similar to that noted in the control heifers or the placebo injected heifers while injection of estradiol 17β decreased the proportion of heifers which became pregnant.In trial 2, 58 heifers which had been bred 1 or 2 times without becoming pregnant were divided into a control group and a group in which heifers received 1000 IU of HCG 96 hr. after observed estrus. In heifers receiving HCG, serum levels of progesterone were higher (P<.01), on day 9 and 16 post estrus than in controls but no difference in serum progesterone was noted (P>.05) between pregnant and non-pregnant heifers on day 9 or 16. The proportion pregnant did not differ (P>.05) between heifers receiving HCG and control heifers.     

A specific progesterone receptor of myometrial cytosol from the rhesus monkey.

A specific progesterone receptor of myometrial cytosol from the rhesus monky is described. Characterization of the receptor by sucrose density gradient centrifugation revealed 2 peaks at 4s and 7.5s. The 4s peak seen in all groups (castrate; castrate plus estrogen treated; castrate plus estrogen and progesterone treated) contained little specific progesterone binding but the 7.5s peak, seen only in the estrogen-treated animal, was specific for progesterone. Competition studies revealed the reeceptor affinities to be: progesterone 100, 5alpha dehydroprogesterone 81.9 melengestrol acetate 72.5, norgestrel 53, desoxycorticosterone 25.9, 5beta-dihydroprogesterone 1.2, and 17 hydroxyprogesterone less than 1. Receptor levels measured from Scatchard plot analysis of of equilibrium data were 7 fM/mg cytosol protein (castrate), 45.2 fM/mg (p less than .01, estrogen treated), and 10.5 (estrogen plus progesterone treated). The association constant (approximately 5 x 10(-9)M) was similar in all 3 groups.     

Progesterone maintenance of the placental progesterone receptor and placental growth in ovariectomized rats

These studies examine the trophic effects of progesterone (P) on the progesterone receptor (Rp) and growth of the decidua basalis (DB) and junctional zone (JZ) in the rat placenta. Pregnant rats were ovariectomized (Ovx) in mid-pregnancy and received steroid replacement therapy consisting of implantation of P pellets (25 mg) and injections of estradiol (E), 2 micrograms s.c., daily. Placental protein synthesis, measured by 3H-leucine incorporation in vitro, decreased more than 99% within 24 h of Ovx. However, treatment with P immediately after castration maintained control levels of synthesis. Delay of P treatment for 4 h caused a 60% decline in protein production measured 20 h later (p less than 0.01). Intraperitoneal implantation of a 50-mg pellet of the antiprogestin, RU-38486, in intact pregnant rats decreased protein synthesis by 50% within 6 h and by more than 90% 12 h and 24 h post-implantation (p less than 0.01). Growth of DB and JZ in Ovx rats treated for 48 h with P and/or E was studied both histologically and by changes in protein and DNA content. Rp binding activity was also measured by exchange assay under equilibrium conditions. Only P was able to reverse the effects of Ovx on growth of the DB and JZ. P also maintained Rp levels in the DB above those observed in Ovx and Ovx + E-treated groups (p less than 0.01). The Rp may be a constitutive product in the JZ since binding activity was not altered by Ovx or by steroid treatments. This study shows that P is clearly a trophic hormone of the maternal and chorioallantoic placenta and is essential for placental growth, cellular differentiation, and histological integrity.     

Estradiol-induced progesterone receptor synthesis in normal and diabetic ovariectomized rat uterus

Estrogen stimulation of progesterone-receptor (Prog R.) synthesis is an important parameter of the sex hormones activity at the uterine level. Experimental diabetes in the rat has been shown to perturb protein synthesis in some tissues and to reduce, under certain circumstances, estrogen and androgen activity on their respective target tissues. The present work tended to evaluate the effect of streptozotocin diabetes on estradiol (E2) stimulation of Prog. R and on Prog R. kinetics in the rat uterus. Two groups of diabetic rats were primed for three consecutive days with 5 microg. E2 s.c. (EP). One group received an acute i.p. injection of progesterone (P), 1 h before sacrifice (Inj), the other group did not (n Inj). Two other groups, not primed with E2 (nEP) were similarly injected or not with P. Four groups of non diabetic animals served as controls. Estrogen priming induced a 20-25% increase in DNA content, both in controls and in diabetics. Protein content was also increased to almost the same extent in diabetics and controls; protein concentration remained however slightly lower in cytosol of EP diabetics as compared to controls. Prog R. increased about 7-fold in cytosol and 4-5-fold in nuclei of EP control and diabetic groups. Cytosol to nuclei ratios of Prog R. decreased similarly in Inj. EP diabetics and controls, compared to the corresponding n Inj. groups. It is concluded that estrogen priming stimulated Prog R., total protein and DNA synthesis to the same extent in diabetic as in control rats Prog R. kinetics was unaltered in diabetics. This finding might be relevant to situations like early pregnancy, when Prog R. levels change rapidly and specifically in relation with the time and the site of implantation.     

Estradiol and progesterone differentially regulate formalin-induced nociception in ovariectomized female rats

Clinical and preclinical studies have found sex-specific differences in the discrimination and perception of inflammatory stimuli. The emerging picture suggests that the biological basis of these differences resides in the regulatory activity of gonadal hormones in the central nervous system. This study describes the effects of ovarian hormones in inflammatory pain processes. Ovariectomized rats received estradiol and/or progesterone, and the number of paw flinches was measured after 1, 2.5 or 5% formalin administration. Both estradiol and progesterone altered the number of flinches only after 1% formalin administration. Estradiol significantly reduced the overall number of flinches during Phase II of the formalin nociceptive response while progesterone attenuated Phase I of the response. After co-administration of estradiol and progesterone, progesterone reversed estradiol's analgesic effect in Phase II, however, estradiol did not reverse progesterone's analgesic activity in Phase I. To determine if estradiol effects are receptor-mediated, tamoxifen (selective estrogen receptor mediator, 15 mg/kg) or alpha-estradiol (an inactive isomer of estradiol, 20 microg) were utilized. Tamoxifen decreased the number of formalin-induced flinches during Phase II while alpha-estradiol did not affect any formalin-induced responses. When co-administered with estradiol, tamoxifen failed to reverse estradiol's effect, suggesting both tamoxifen and estradiol activate similar intracellular mechanisms. Although Western blot analysis detected the presence of estradiol alpha and beta and progesterone B receptors in the spinal cord, hormone replacement treatments had no effects on the levels of these receptors. We postulate that the mechanisms by which estradiol and progesterone induce analgesia occur through the activation of their receptor at the spinal cord level.     

Binding characteristics of progesterone and antiprogestin ZK 98.299 in human endometrial and myometrial cytosol

The binding of ZK 98.299, a synthetic progesterone antagonist, with human endometrium and myometrium cytosol was studied and compared with that of progesterone. Progesterone showed specific saturable binding to its receptors in both endometrium and myometrium. ZK 98.299 and progesterone were mutually competitive for binding to progesterone receptors; however, the relative binding affinity of ZK 98.299 was 16% that of progesterone. ZK 98.299 exchanged the progesterone-labelled receptor sites. [3H]ZK 98.299 showed specific binding which was linearly related to the cytosol protein concentration. The binding was not saturable at 15 nM of ligand. The binding capacity and binding affinity of ZK 98.299 receptor was less than that of progesterone. Progesterone also partially displaced the binding of [3H]ZK 98.299. This study suggest that ZK 98.299 and progesterone both bind to the same protein. However, whether ZK 98.299 binds to progesterone receptors alone or even to other functionally related sites is not known. It appears that ZK 98.299 when present in higher concentration than progesterone would be an effective receptor ligand.     

Relaxin in the male

Relaxin, a hormone usually associated with pregnancy, has been found in the semen plasma of many species. The prostate gland appears to be a source of relaxin. Relaxin stimulates sperm motility from suboptimal samples and increases sperm penetration into oocytes. Thus, relaxin may be an effective therapeutic agent in male infertility.     

Effects of estrogen and progesterone on uterine sialic acid in ovariectomized rats

The effects of estradiol-17β and progesterone on uterine sialic acid of ovariectomized rats have been examined. In contrast to a previous report, progesterone was found in two of three experiments of different design to increase uterine sialic acid concentration above that produced by estradiol-17β alone; in the third experiment, it had no significant effect. This effect of progesterone was independent of the duration of treatment with exogenous hormones or of whether or not uterine luminal fluid was removed by blotting before assaying sialic acid. In a factorially designed experiment with four levels of estradiol-17β and three of progesterone, a dose-response relationship was found between estradiol-17β, but not progesterone, and uterine sialic acid concentration. It is concluded that, in some circumstances, estrogen and progesterone can act synergistically to increase uterine sialic acid concentration.     

The effect of progesterone on the activity-wheel running of ovariectomized rats

Progesterone treatment failed to significantly depress activity-wheel activity elicited by food deprivation in either ovariectomized or ovariectomized estrogen-treated rats. These results indicate: (1) that the lack of effect of progesterone on the activity of ovariectomized rats is not due only to their already low level of activity; and (2) that the depressant effect of progesterone on activity in the estrogen-treated ovariectomized rat is, to a large extent, a specific inhibition of estrogen facilitation of activity, rather than a nonspecific activity depressant effect.     

Effects of diosgenin on myometrial matrix metalloproteinase-2 and -9 activity and expression in ovariectomized rats

Diosgenin, a traditional Yam extraction, has been used in hormone replacement for menopausal women. We aimed to investigate the influences of diosgenin administration upon the MMP-2 and -9 activity and expression and reproductive hormones of ovariectomized (OVX) rats, a model of menopausal status. Seven-week old female Wistar rats with bilateral OVX or sham operation (controls) were divided and administered different dosages of diosgenin (0, 10, 50, or 100 mg/kg/day) for 8 weeks. Serum was then sampled for progesterone (P4) and estradiol (E2) assay and uterine horns harvested. Myometrial MMP-2 and -9 activity and expression were surveyed and myometrial collagen expression was also assayed. The results show higher body weight in OVX rats across the 8 weeks post surgery and no significant differences were noted among OVX or Sham rats with diosgenin supplements. There were lower P4 and E2 concentrations in OVX rats compared to Sham rats, and higher P4 concentration of Sham rats post diosgenin supplement. MMP-2 and -9 mRNA expression and activity was lower in OVX rats, although higher MMP-2 and lower MMP-9 activity/mRNA expression was observed in OVX rats post diosgenin supplementation. Collagen mRNA expression was higher in OVX rats compared to Sham controls, and diosgenin administration decreased collagen mRNA expression in OVX rats. In conclusion, diosgenin is associated with gelatinase expression and collagen metabolism in OVX rats. Diosgenin administration can partially reverse the effects of OVX upon MMP functions and hormone status. Adequate diosgenin supplement might modulate myometrial gelatinase expression and collagen metabolism in menopausal subjects.