Prostaglandin E2 levels in uterine tissues and its relationship with uterine and luteal progesterone during the estrous cycle in dairy cows

Endometrial tissue homogenates obtained at luteal and follicular stages of the estrous cycle were determined for prostaglandin E(2) and progesterone contents by EIA and RIA, respectively. In Experiment 1, the concentrations and changes of PGE(2) in uterine tissues collected by biopsy before slaughfter and subsequent samples collected at 30, 60 and 90 min after slaughter were measured. No significant differences were observed in the concentration of PGE(2) preslaughter or at 30 and 60 min post slaughter. However, there was a significant decrease (P<0.01) in PGE(2) concentration 90 min post slaughter. In Experiment 2, the concentrations of PGE(2) in the ipsilateral and contralateral horns in relation to corpus luteum function were compared. A significant (P<0.05) interaction was found between stages of estrous cycle (luteal vs follicular) based on CL progesterone content, and type of uterine horn (ipsilateral vs contralateral) on uterine PGE(2) levels. The PGE(2) concentration was significantly higher (P<0.01) at luteal phase than at follicular phase. During the luteal phase PGE(2) concentrations in tissues of the uterine horn ipsilateral to the corpus luteum was significantly higher (P<0.01) than the contralateral horn. The PGE(2) concentration was low and did not differ significantly between horns during follicular phase. A parallel increase (luteal: high) and decrease (follicular: low) in PGE(2) and progesterone concentrations were observed. Correlations were observed for CL progesterone and uterine PGE(2) concentrations as well as for PGE(2) and progesterone concentrations in uterine tissues (r=0.70 and r=0.60, respectively). The results show that the increase in PGE(2) concentrations in uterine tissues coincides with the high uterine progesterone concentrations during luteal phase.     

Morphology of the genital organs of the female red-rumped agouti (Dasyprocta leporina,Linnaeus, 1758) during estrous cycle phases and in advanced pregnancy

The study investigated the gross and microscopic anatomy of the genital organs of 20 agoutis at different stages of the estrous cycle and four in the final trimester of pregnancy. Specimens were euthanized and their reproductive organs were fixed in a 4% paraformaldehyde or 2.5% glutaraldehyde solution and submitted to routine histological techniques for light and scanning electron microscopy. In the ovary, during the proestrus phase, we observed developing follicles and corpus luteum (CL) in regression; during estrus, there were Graafian follicles; during metestrus, there was a hemorrhagic corpus, whereas in diestrus, there was a mature CL. The uterus was partially double because the cervix was cranially septate but caudally, the septum disappeared, forming a single ostium that opened into the vagina. Changes occurred along the estrous cycle in the uterine and vaginal epithelia, that is, an increase in the uterine epithelium height accompanied by an increase of thickness of the vaginal epithelium during the follicular phase and a decrease of thickness of both epithelia during the luteal phase. The endometrial lining was composed of a simple cuboidal epithelium to simple columnar epithelium with basal nuclei. The vaginal mucosa consisted of epithelium that varied from nonkeratinized stratified squamous (luteal phase) to keratinized stratified squamous (follicular phase). The clitoris was external to the vagina. It presented two protruding lateral keratinized spicules and a centralized urethra, with no common parts between the urinary and genital tracts. Anatomical and histological changes were observed mainly in the cervix, vagina and spicules of the clitoris during the EC.     

Local vascular pathway for progesterone transfer to the brain after nasal administration in gilts

In the present study we examined whether local transfer of intranasally administrated tritiated progesterone (3H-P4) would increase its concentration in blood supplying the brain and hypophysis in comparison with other organs. Additionally, the effect of estrous cycle on the P4 transfer was evaluated on isolated gilts' heads. In the first experiment 3H-P4 was instilled into the nasal cavities of anaesthetized, immature pigs (n=10). Simultaneous blood samples were collected for radioactivity measurement every minute from the same occluded carotid artery through two catheters; one catheter was pointed towards the head, the other one towards the heart. In eight animals the ratio calculated between the 'head' and 'heart' samples was significantly (p<0.05) higher than 1 and reached a mean (+/- SEM) level of 3.23 +/- 0.81. In two animals a much higher ratio was observed. A head/heart ratio>1 indicates an existence of local transfer of 3H-P4 from venous blood to the carotid blood. In the second experiment, heads of 26 mature, cycling gilts were perfused through the right carotid artery with autologous blood. The outflow from the left carotid artery was collected as 1 min samples. 3H-P4 was infused into the angularis oculi veins. Transfer of 3H-P4 from the venous blood into the arterial blood reached the mean (+/- SEM) level of 4.11 +/- 1.08 pg/ml on days 2-4, 3.2 +/- 0.70 on days 17-21 and 0.94 +/- 0.22 pg/ml on days 15-16 of the estrous cycle. No 3H-P4 transfer was observed on days 9-11. These findings demonstrate that nasally administered progesterone can reach the brain in locally higher concentration through the vascular pathway. Moreover, the between-vessel transfer of P4 is significantly affected by the stage of the estrous cycle.     

The influence of reproductive phase on lipopolysaccharide stimulation of prostacyclin production and cyclooxygenase-2 protein concentrations in reproductive and systemic arteries of the sheep

Cyclooxygenase, the enzyme that converts arachidonate to prostaglandins, plays a regulatory role in vasodilation under normal and pathological conditions. Studies were conducted to determine the effects of reproductive phase and lipopolysaccharide (LPS) on production of PGI2 and amounts of cyclooxygenase protein in uterine, mammary, mesenteric, and renal arteries. Arteries were collected from ewes during the follicular (Day 0 = estrus) or luteal (Day 10) phase of the estrous cycle and were cultured in the presence of LPS. After 24 h, media were collected and analyzed for 6-keto-PGF1alpha, the stable metabolite of PGI2. In addition, arteries were collected and homogenized and the relative concentration of cyclooxygenase was determined via Western analysis. Lipopolysaccharide stimulated PGI2 production in all four-artery types from both follicular and luteal phase ewes (p < 0.001). Upon LPS stimulation, uterine and mammary arteries produced more PGI2 compared to mesenteric and renal arteries (p = 0.04). The phase of estrous cycle did not affect PGI2 production by any of the artery populations exposed to LPS (p = 0.35). There was no cyclooxygenase-2 in untreated uterine and mammary arteries and no cyclooxygenase-2 was detected in untreated or LPS-treated mesenteric and renal arteries. In contrast, LPS-treated uterine and mammary arteries from luteal phase ewes had higher (p = 0.064) cyclooxygenase-2 concentrations than those from follicular phase ewes. These results suggest that the hormone conditions of the follicular (high estrogen) and luteal (high progesterone) phases of the ovarian cycle play a role in regulating uterine and mammary artery but not mesenteric and renal artery response to LPS.     

Functional anatomy of the female genital organs of the wild black agouti (Dasyprocta fuliginosa) female in the Peruvian Amazon

This study examined anatomical and histological characteristics of genital organs of 38 black agouti females in the wild in different reproductive stages, collected by rural hunters in the North-eastern Peruvian Amazon. Females in the follicular phase of the estrous cycle had greater antral follicle sizes than other females, the largest antral follicle measuring 2.34mm. Antral follicles in pregnant females and females in luteal phase of the estrous cycle had an average maximum diameter smaller than 1mm. In black agouti females in follicular phase, some antral follicles are selected to continue to growth, reaching a pre-ovulatory diameter of 2mm. Mean ovulation rate was 2.5 follicles and litter size was 2.1 embryos or fetuses per pregnant female, resulting in a rate of ovum mortality of 20.8%. Many follicles from which ovulation did not occur of 1-mm maximum diameter luteinize forming accessory CL. The constituent active luteal tissues of the ovary are functional and accessory CL. Although all females had accessory CL, transformation of follicles into accessory CL occurred especially in pregnant females, resulting in a contribution from 9% to 23% of the total luteal volume as pregnancy advances. The persistence of functional CL throughout pregnancy might reflect the importance for the maintenance of gestation and may be essential for the continuous hormonal production. The duplex uterus of the agouti female is composed by two completely independent uterine horns with correspondent separate cervices opening into the vagina. In pregnant females, most remarkable observed uterine adaptations were induced by the progressive enlargement caused by the normal pregnancy evolution. The wild black agouti showed different vaginal epithelium features in accordance with the reproductive state of the female.     

Increase in progesterone and luteal blood flow without a luteolytic response after prostaglandin F2α treatment in early luteal-phase heifers

Acute changes in circulating progesterone concentration and luteal blood flow in heifers after a conventional dose of prostaglandin F(2α) (PGF; 25mg dinoprost, i.m.) were compared between the early luteal phase (Day 3) and midluteal phase (Day 10; Day 0=ovulation), using four groups (Day-3 control, Day-3 PGF, Day-10 control, and Day-10 PGF; n=6 heifers/group). Blood samples were collected at 0, 2, 5, 10, 15, 30, 60, and 120 min (0 min=treatment). Percentage of luteal area with color-Doppler blood-flow signals was estimated at 0, 10, and 30 min. In the Day-3 and Day-10 PGF groups, progesterone increased to a peak at 15 min. In the Day-3 PGF group, progesterone decreased to the pretreatment concentration by 60 min but did not decrease to below the pretreatment concentration during the 2-h experimental period. In the Day-10 PGF group, progesterone decreased to below pretreatment concentration by 30 min, indicating a luteolytic response. In the Day-3 and Day-10 PGF groups, luteal blood flow increased within 10 min and remained elevated until the last examination at 30 min. The absence of a decrease in progesterone to below pretreatment concentrations in the Day-3 PGF group indicated that luteolysis does not necessarily follow a transient increase in progesterone and a concomitant increase in luteal blood flow. The immediate transient increase in progesterone and an increase in luteal blood flow without a subsequent decrease in progesterone to below pretreatment concentrations after PGF treatment in early luteal-phase heifers are novel findings.     

Prolongation of the luteal phase by prostaglandin E(2) during the estrous cycle in the cow. A preliminary report

An experiment was conducted to determine the effects of prostaglandin E(2) (PGE(2)) on ovarian progesterone secretion during the estrous cycle in the cow. Intraluminal uterine catheters were implanted in three beef cows (2 treated, 1 control), and 1.3 mg of PGE(2) were infused into the uterus every 4 hours from days 9 through 21 post-estrus. Blood samples were collected from the jugular vein at 2-hour intervals from days 9 to 21 and twice daily from day 22 to 28 post-estrus. Progesterone was measured by applying a specific, direct plasma radioimmunoassay in all samples without extraction. Intrauterine infusion with PGE(2) resulted in maintenance of luteal-phase progesterone secretion until day 21 post-estrus, 4 days after luteal regression occurred in the vehicle-treated cow. In this study, we demonstrated that PGE(2) can prolong the presence of luteal phase plasma progesterone concentrations by possibly stimulating in vivo steroidogenesis by the corpus luteum during the estrous cycle in the cow.     

Counter current transfer and back transport of 3H-PGF2 alpha in the cow's broad ligament vasculature ipsilateral and contralateral to the corpus luteum

Eight cows of similar age (5-7 years) were chosen for the experiment. Isolated reproductive tract was supplied with autologous oxygenated and heated (40 degrees C) blood through the uterine artery and ovarian artery. 3H-PGF2 alpha in total dose of 2 MBq (10(7) cpm) was injected into each of the uterine lumen of isolated organ. Blood samples were collected at 5 min intervals during 120 min of experiment using cannulae inserted into the branches of uterine arteries about 1 cm below the horns and from ovarian arteries inserted 0.5 cm below the ovaries. The concentration of 3H-PGF2 alpha found in blood plasma taken from uterine artery or from ovarian artery on the side with active corpus luteum (CL) was significantly lower (p less than 0.001) compare with contralateral side to active CL. Radioactive PGF2 alpha found in branches of uterine arteries on both ipsilateral and contralateral side to CL was significantly higher (p less than 0.001) compare to ovarian artery of the same side. It is concluded that absorption of 3H-PGF2 alpha from uterine lumen into venous blood as well as its counter current transfer in area of broad ligament vasculature were reduced on the side of uterine horn with active CL probably as an effect of estrogen:progesterone ratio on vascular constriction in area of uterine vasculature.     

Effect of exogenous gonadal steroids and pregnancy on uterine luminal prostaglandin F in mares

Two experiments were conducted to assess the effect of exogenous hormone treatment on uterine luminal prostaglandin F (PGF). In the first experiment ovariectomized pony mares received either corn oil (21 days, n = 3), estradiol valerate (21 days, n = 3), progesterone (21 days, n = 3) or estradiol valerate (7 days) followed by progesterone (14 days, n = 4). Progesterone treated mares had higher (P<.01) uterine luminal PGF compared with all other groups, and no differences were detected between other treatment comparisons. In Experiment II, uterine fluid was collected from 4 ovariectomized horse mares before and after treatment with estradiol valerate (7 days) followed by progesterone (50 days). Pretreatment uterine luminal PGF levels were lower (P<.001) than post-treatment levels (.03 vs 76.80 ng/ml). In a third experiment PGF was measured in uterine fluid of pony mares on days 8, 12, 14, 16, 18 and 20 of the estrous cycle and pregnancy. In nonpregnant mares a day effect P<.03) was observed in which uterine fluid PGF increased during the late luteal phase and declined thereafter. In contrast, no day effect was observed in pregnant animals and uterine luminal PGF was lower (P<.001) than in cycling animals. These studies indicate that exogenous progesterone administration results in a large increase in uterine luminal PGF, whereas, pregnancy results in suppression. Taken collectively with previous work from our laboratory, these results suggest that while the endometrium of pregnant mares is capable of producing large amounts of PGF, the presence of a conceptus impedes its synthesis and/or release which allows for luteal maintenance.     

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.     

The effect of oestradiolum benzoicum and progesterone on the noradrenalin content in organs of the female reproductive system of sexually immature pigs

The paper describes the effect of exogenous oestradiol and progesterone on noradrenalin (NA) content in the ovary, tuba uterina, uterus, and vagina, and on the morphological features of adrenergic nerves in these organs in sexually immature female pigs. 30 animals were used, of body weight 37 to 43 kg, and age 115 to 125 d. The animals were divided randomly into 4 groups. Group I (n = 6) constituted the control, group II (n = 8) was given oestradiolum benzoicum, group III (n = 8) was given progesterone, and group IV (n = 8) was given oestradiol and progesterone. Oestradiol injections resulted in a 1.5-fold increase of NA content in the ovary, a 4.5-fold increase in the tuba uterina, and a decrease of NA content in the uterine horn, cervix, and in the vagina, to 0.6 of the control NA level. Progesterone injections did not cause changes in NA content in the ovary, while there was 8.5-fold increase of NA in the oviduct, a 4-fold increase in the uterile horn, a 6-fold increase in the cervix, and a 3-fold increase in the vagina compared to the control. Joint injections of oestradiol and progesterone resulted in a decrease of NA content in the ovary to about 0.5 of the control level, and in an increase of NA content of about 5-fold in the tuba uterina, 7-fold in the uterine horn, 8-fold in the cervix, and 2-fold in the vagina. The increase of NA content in the organs was coupled with a slight increase in the density of the adrenergic innervation and the intensity of fluorescence of these nerves compared to the control. The results were compared to the results of similar but not identical studies, carried out on other mammals.     

The effect of oestradiolum benzoicum and progesterone on AChE activity in the nerves of the female reproductive system of immature pigs

The present paper describes the effect of exogenous oestradiol and progesterone on the AChE activity in nerves of the ovary, oviduct, uterus, and vagina, and on morphological features of the AChE-positive nerves of these organs in sexually immature pigs. Studies were carried out on 30 sexually immature pigs, 37 to 43 kg in weight, at the age of 115 to 125 d, of the BWP (Big White Polish) race. The animals were divided into 4 groups. Group I (n = 6) constituted the control. Animals in group II (n = 8) received oestradiolum benzoicum. Those in group III (n = 8) were given progesterone, and in group IV (n = 8) oestradiol and progesterone. Oestradiol injections resulted in a 1.2-fold increase of AChE activity in the ovary, 1.3-fold in the oviduct, 1.7-fold in the uterine horn, and a 1.2-fold decrease of this activity in the cervix, while no differences were noted in the vagina. Progesterone injections did not affect AChE activity in the ovary, while a 1.1-fold increase was observed in the oviduct, 1.2-fold in the uterine horn and 1.1-fold in the cervix compared to the control. A 1.1-fold decrease of this activity was observed in the vagina. Joint injections of oestradiol and progesterone resulted in about a 1.4-fold increase of AChE activity in the ovary, 1.3-fold in the oviduct, 1.4-fold in the uterine horn, 1.6-fold in the vagina, while no changes were observed in the cervix. The results suggest the effect of the administered hormones upon AChE activity in the organs under study, mainly in the oviduct, uterine horn, and vagina. They also point to the possibility of a synergic effect of oestradiol and progesterone as regards to an increase of the activity.     

Role of prostaglandin E2 in basal and noradrenaline-induced progesterone secretion by the bovine corpus luteum

The role of prostaglandin E2 (PGE2) in basal and noradrenaline (NA)-stimulated utilization of high density lipoprotein (HDL) as a source of cholesterol for progesterone synthesis was examined. In Experiment 1, a cannula was inserted into the aorta abdominalis through the coccygeal artery (cranial to the origin of the ovarian artery) in mature heifers, to facilitate infusion of NA (4 mg/30 min; n = 3) on day 10 of the estrous cycle. Three other heifers were similarly cannulated to serve as control. Before, during, and after NA or saline infusion, blood samples from the vena cava were collected every 5-15 min for analysis of PGE2, progesterone, and cholesterol. Each NA infusion stimulated (P < 0.01) secretion of both hormones in heifers. Short-duration increases (P < 0.05) in progesterone were observed due to the infusion of NA while cholesterol was not altered significantly. In addition, increases in PGE2 concentrations (P < 0.05) compared to controls were seen after NA infusion. Therefore, we used an in vitro model to verify the effect of PGE2 on HDL utilization by luteal cells from day 5 to 10 of the estrous cycle. In the preliminary experiment, 10(-6) M of PGE2 out of four different doses examined was selected for further studies, since it evoked the highest release of progesterone. In the next experiment, it was found that HDL increases progesterone secretion by luteal cells and both PGE2 and LH increased (P < 0.05) the response to HDL while NA did not. In the last in vitro experiment, progesterone stimulated PGE2 secretion by luteal cells. In conclusion, PGE2 may be directly involved in the utilization of cholesterol from HDL for progesterone synthesis. Furthermore, PGE2 may influence NA-stimulated progesterone secretion by the corpus luteum (CL). It is concluded that there is a positive feedback loop between progesterone and luteal PGE2 during days 5-10 of the estrous cycle.     

Blood flow to the genital tract of oestrous and dioestrous guinea-pigs

Ovarian, uterine and vaginal blood flow were determined in 22 virgin guinea-pigs by the tracer microsphere technique. Measurements were made during oestrus, when cornified cells appeared in the vaginal smear (Day 1), or during the luteal phase of the cycle (Day 11). The total rate of blood flow to the genital tract was 0-58 ml.min-1 on Day 11 and 2-92 ml.min-1 on Day 1. This difference was largely due to an 8-fold increase in uterine blood flow from 0-26 to 2-01 ml.min-1. Although uterine weight increased over the same period, there was a significant increase in uterine tissue perfusion from 0-32 to 1-18 ml.min-1.g-1. The vagina exhibited a similar pattern, including a significant increase in tissue perfusion. Ovarian blood flow decreased from a value of 0-19 ml.min-1 during the luteal phase to 0-10 ml.min-1 at oestrus. Perfusion of the ovarian tissue was considerably greater on Day 11 than on Day 1 (2-86 versus 1-39 ml.min-1.g-1).     

Ovarian and placental production of progesterone and oestradiol during pregnancy in reindeer

We obtained uterine and peripheral venous plasma, and samples of luteal and placental tissues from 2- to 7-year-old, Eurasian mountain reindeer (Rangifer tarandus tarandus) from a free-living, semi-domesticated herd in northern Norway in November 1995, and February and March 1996. In November, ovarian venous blood was also collected from four animals. Plasma samples were assayed for progesterone and oestradiol. The tissue samples were examined by light and electron microscopy, steroid dehydrogenase histochemistry, and northern blot analysis for RNAs for 3beta-hydroxy-steroid dehydrogenase (3beta-HSD) and P450 (side chain cleavage (scc)). Peripheral blood was taken from non-pregnant females in the same herd on the same dates. Peripheral progesterone concentrations in pregnant reindeer (3.4 +/- 0.5 ng/ml, n = 8) clearly exceeded those in non-pregnant animals (0.40 +/- 0.14 ng/ml; P < 0.0004 , n = 10) but oestradiol levels were only marginally higher in pregnant (6.0 +/- 0.7 pg/ml) than in non-pregnant (4.8 +/- 0.5 pg/ml; P = 0.35) reindeer at the stages examined. In pregnant animals, peripheral progesterone and oestradiol concentrations rose slightly between November and March but the differences did not reach significance (progesterone, P = 0.083; oestradiol, P = 0.061). In November, progesterone concentrations in the ovarian vein (79 +/- 15 ng/ml) greatly exceeded (P < 0.03) those in the uterine vein ( 10 +/- 4 ng/ml) which in turn exceeded the levels in the peripheral blood (2.8 +/- 0.4 ng/ml; P < 0.29). Oestradiol concentrations were slightly but significantly (P < 0.05) higher in the ovarian (20 +/- 3 pg/ml) than the uterine vein (13 +/- 1 pg/ml) and, in turn, greater (P < 0.03) than in peripheral blood (4.6 +/- 0.4 pg/ml). All samples of luteal tissue consisted exclusively of normal fully-differentiated cells and stained intensely for 3beta-HSD. Isolated groups of placental cells also stained strongly for 3beta-HSD. RNA for P450 (scc) and 3beta-HSD was abundant in all corpora lutea and lower concentrations of P450 (scc) were present in the placenta. 3beta-HSD RNA in the placenta was below the limit of detection. We conclude that the corpus luteum remains an important source of progesterone throughout pregnancy in reindeer but that the placenta is also steroidogenic.     

Development of corpus luteum susceptibility to an analog of prostaglandin F2α, throughout the luteal phase in llamas (Lama glama)

The aim of the present study was to evaluate the susceptibility of the corpus luteum to d-cloprostenol (synthetic analog of PGF(2α)) throughout the luteal phase in llamas. Female llamas (n=43) were induced to ovulate by GnRH injection in the presence of an ovulatory follicle and randomly assigned into one of six groups: control and treated with an injection of d-cloprostenol on Day 3, 4, 5, 6 or 8 post GnRH. Blood samples were collected to determine plasma progesterone concentrations. There was no effect of treatment on animals injected on Day 3 or 4 post-GnRH. In animals treated on Day 5, different responses were observed. No effect of treatment was recorded in 27% of the animals whereas 55% of the llamas showed a transitory decrease followed by a recovery in plasma progesterone concentrations after d-cloprostenol injection, indicative of a resurgence of the corpus luteum, extending the luteal phase a day more than in control animals. In the remaining 18% of the animals injected on Day 5, (corresponding to those exhibiting the greatest plasma progesterone concentrations at the day of injection), complete luteolysis was observed. Plasma progesterone concentrations decreased to below 1 ng ml(-1) 24 h after d-cloprostenol in llamas injected on Day 6 or 8 post-GnRH. In conclusion, the corpus luteum of llamas is completely refractory to PGF(2α) until Day 4 after induction of ovulation, being partially sensitive by Day 5 and fully responsive to PGF(2α), by Day 6 after induction of ovulation.     

Endothelial vasodilator production by uterine and systemic arteries. IV. Cyclooxygenase isoform expression during the ovarian cycle and pregnancy in sheep

Uterine artery endothelial production of the potent vasodilator, prostacyclin, is greater in pregnant versus nonpregnant sheep and in whole uterine artery from intact versus ovariectomized ewes. We hypothesized that uterine artery cyclooxygenase (COX)-1 and/or COX-2 expression would be elevated during pregnancy (high estrogen and progesterone) and the follicular phase of the ovarian cycle (high estrogen/low progesterone) as compared to that in luteal phase (low estrogen/high progesterone) or in ovariectomized (low estrogen and progesterone) ewes. Uterine and systemic (omental) arteries were obtained from nonpregnant luteal-phase (LUT; n = 10), follicular-phase (FOL; n = 11), and ovariectomized (OVEX; n = 10) sheep, as well as from pregnant sheep (110-130 days gestation; term = 145 +/- 3 days; n = 12). Endothelial and vascular smooth muscle (VSM) COX-1 protein levels and uterine artery endothelial cell COX-1 mRNA levels were compared. Using immunohistochemistry and Western analysis, the primary location of COX-1 protein was the endothelium; that is, we observed 2.2-fold higher COX-1 protein levels in intact versus endothelium-denuded uterine artery and a 6.1-fold higher expression in the endothelium versus VSM (P < 0.05). COX-2 protein expression was not detectable in either uterine artery endothelium or VSM. COX-1 protein levels were observed to be higher (1.5-fold those of LUT) in uterine artery endothelium from FOL versus either OVEX or LUT nonpregnant ewes (P < 0.05), with substantially higher COX-1 levels seen in pregnancy (4.8-fold those of LUT). Increases in uterine artery endothelial COX-1 protein were highly correlated to increases in the level of COX-1 mRNA (r(2) = 0.66; P < 0.01) for all treatment groups (n = 6-8 per group), suggesting that increased COX-1 protein levels are regulated at the level of increased COX-1 mRNA. No change in COX-1 expression was observed between groups in a systemic (omental) artery. In conclusion, COX-1 expression is specifically up-regulated in the uterine artery endothelium during high uterine blood flow states such as the follicular phase and, in particular, pregnancy.     

Short and long phases of progesterone secretion during the oestrous cycle of the African elephant (Loxodonta africana)

Serum samples were collected from 3 mature female African elephants once each week for 15-18 months. Circulating concentrations of progesterone, oestradiol and LH were determined by radioimmunoassay (RIA). The LH RIA was validated by demonstrating parallel cross-reaction with partly purified elephant LH pituitary fractions. Changing serum progesterone concentrations indicated an oestrous cycle length of 13.3 +/- 1.3 weeks (n = 11). The presumed luteal phase, characterized by elevated serum progesterone values, was 9.1 +/- 1.1 weeks (n = 11). Two abbreviated phases of progesterone in serum lasting 2-3 weeks were observed in 2 elephants, indicating short luteal phases. Oestradiol concentrations in serum were variable, with no clear pattern of secretion. More frequent blood samples were collected during periovulatory periods and 9 distinct LH peaks were detected; all were followed by rises in serum progesterone concentrations. Periovulatory changes in progesterone and LH in sera correlated with external signs of oestrus and mating behaviour.     

Salivary measurement of episodic progesterone release

Episodes of luteal progesterone release which have been identified from serum samples can also be observed in saliva. Components of these release episodes--amplitude, duration, and frequency--can be separated and characterized. In a sample of 16 cycle days collected from 6 normal women, pulse duration and frequency increased significantly from the early to the mid-luteal phase, and then decreased by the late luteal phase (2-4 days before menses). Pulse amplitude also increased early to mid-luteal, but continued to increase significantly into the late luteal, and did not decline until the peri-menstrual phase (0-1 days before menses). Circadian variation, previously considered unimportant for progesterone, was also observed in late luteal samples. Extension of studies of ultradian release characteristics to groups known to vary from controls in broad-scale progesterone patterns, such as young, nutritionally stressed, or exercising women, should help to elucidate the potential functional significance of fine-grained release components. Such knowledge can also inform protocols of anthropological studies which seek to identify broad progesterone patterns or to classify and compare reproductive status.     

Reproductive functional anatomy and oestrous cycle pattern of the female brush-tailed porcupine (Atherurus africanus,Gray 1842) from Gabon

In the present study, we examined certain features of the functional anatomy of the female genital tract of the wild brush-tailed porcupine (Atherurus africanus) to obtain data on the reproductive biology of this African forest rodent. Two consecutive experiments were performed. The aim of the first was to establish macroscopic and microscopic features of the genital organs, and to explore correlations between predominant ovarian structures and vaginal contents in 20 wild, mature females. In the second experiment, we inspected the external genitalia and vaginal smears of a further 10 females in captivity on a daily basis for 90 days. The uterus of the brush-tailed porcupine is bicornuate and composed of two separated uterine horns, a uterine body and cervix. The genital tract does not present a vaginal vestibule. Thus, there is no portion common to genital and urinary tracts. Females in the follicular phase of the oestrous cycle showed increased cornification of the vaginal epithelium and a high density of eosinophilic cells in vaginal smears. The vulva and vaginal opening were open, reddish and tumefacted. In luteal phase or in pregnancy, epithelial cornification and eosinophilic features were notably reduced and the vagina presented a pale, non-tumefacted vulva and a vaginal closure membrane. Females in captivity showed spontaneous cycles, a polyoestrous reproduction pattern and, based on features of the external genitalia and vaginal smears, their oestrous cycle length was 27.1+/-6.4 days (n=12).