Changes in equine endometrial oestrogen receptor alpha and progesterone receptor mRNAs during the oestrous cycle, early pregnancy and after treatment with exogenous steroids

Two experiments were performed to determine changes in the abundance of oestrogen and progesterone receptor (ER alpha and PR) mRNAs in equine endometrium during the oestrous cycle and early pregnancy, and under the influence of exogenous steroids. In Expt 1, endometrial biopsies were obtained from non-mated mares during oestrus and at days 5, 10 and 15 after ovulation, and from pregnant mares at days 10, 15 and 20 after ovulation. There were overall effects of day on the abundance of ER alpha (P = 0.0001) and PR (P = 0.0014) mRNAs. The amount of ER alpha mRNA decreased at day 10 of pregnancy, and PR mRNA was reduced at day 5 in non-mated mares and at day 15 of pregnancy, compared with oestrous values. Experiment 2 was conducted to determine the effects of exogenous steroids on endometrial ER alpha and PR mRNAs. Endometrial biopsies were obtained from 19 anoestrous mares that had been treated with vehicle, oestradiol, progesterone, or oestradiol followed by progesterone for either a short or a long duration. The steroid treatment affected the abundance of ER alpha mRNA (P = 0.0420), which was higher (P < 0.05) in the oestradiol group than in the group treated with oestradiol followed by long duration progesterone. The steroid treatment did not affect the abundance of PR mRNA. These results demonstrate that the amount of steroid receptor mRNA changes with the fluctuating steroid environment in the uterine endometrium of cyclic and early pregnant mares, and that the duration of progesterone dominance may affect ER alpha gene expression. In addition, factors other than steroids may regulate ER alpha and PR gene expression in equine uterine endometrium.     

Effect of plane of nutrition on endometrial sex steroid receptor expression in ewes

The effect of plane of nutrition on progesterone receptor (PR) and estrogen receptor alpha (ERalpha) expression in ovine endometrium was investigated. Rasa Aragonesa ewes (n=26) were fed diets to provide either 1.5 (Group C) or 0.5 (Group L) times the daily maintenance requirement and were slaughtered at Days 5 or 10 of the estrous cycle (Day 0=estrus). PR and ERalpha immunoreactivity were analyzed in eight endometrial cell compartments, defined by cell type and location. Group L had less PR immunostaining on Day 5 (P<0.05), which is consistent with lesser endometrial content of progesterone found in such animals. Most cell types of Group C had down regulation of PR at Day 10, but in Group L, this pattern was observed only in three cell compartments. The lesser PR contents found at Day 5 in Group L ewes may explain the lack of inhibition of PR. No effect of treatment or day of the estrous cycle was observed in ERalpha. Results indicate that endometrial PR is affected in a cell type, in specific manner, by plane of nutrition.     

Concentration of steroids in bovine peripheral plasma during the oestrous cycle and the effect of betamethasone treatment.

Testosterone, oestradiol and progesterone were measured in peripheral plasma during the oestrous cycle of 6 heifers. Oestradiol and progesterone results confirmed earlier reports. Concentration of testosterone on the day of oestrus was 40+/-3 pg/ml (mean+/-S.E.M.), and two peaks were detected during the cycle, one 7 days before oestrus (1809+/-603 pg/ml) and the other (78+/- 7 pg/ml) on the day before the onset of oestrus. The concentration of progesterone declined in most cases 1 day after the maximum concentration of testosterone. Betamethasone treatment in 5 heifers extended luteal function by an average of 10 days: plasma androstenedione and oestradiol concentrations were unaltered; cortisol values were depressed for at least 16 days after treatment; testosterone concentrations were lowered by 13+/-2-4% during treatment, and except in one heifer the peak on Day -7 was abolished.     

Endometrial expression of progesterone receptor and uteroglobin genes during early pregnancy in the rabbit

The progesterone receptor (PR) plays a pivotal role in the maturation process of the secretory endometrium, implantation and maintenance of pregnancy in rabbits. To determine the dynamics of PR gene expression and its physiological significance, the endometrial expression of PR and PR mRNA were evaluated and compared with the expression of the progesterone-regulated uteroglobin (UG) gene during 0–5 days post-coitus in rabbits. The results of immunoblot experiments indicated the presence of PR in endometrial cell extracts from days 1–4 of pregnancy with maximum PR immunostaining on day 2, followed by a marked diminution until its complete disappearance on day 5. When endometrial PR mRNA content was assessed by Northern blots, the results were similar to those of PR immunostaining, with maximal concentrations on the second day after mating. However, PR mRNA levels were still high on day 3, despite the concomitant decrease in immunostainable PR. Endometrial UG gene expression, on the other hand, exhibited a different time sequence. Thus, the UG content in uterine flushings progressively increased from day 3 after mating, reaching maximal levels on the fifth day. The endometrial UG mRNA content presented a similar profile, as its maximum concentration occurred on days 4–5. The overall results indicate that endometrial PR is down-regulated at both the mRNA and protein levels, possibly by endogenous progesterone during early pregnancy. The striking observation that maximal expression of endometrial UG gene products occurred when PR and its mRNA are no longer detectable suggests an important role for this progesterone-binding uterine protein during the preimplantation period. © 1993 Wiley-Liss, Inc.     

Peripheral plasma concentrations of oestradiol, progesterone, cortisol, LH and prolactin during the oestrous cycle in the cow, with emphasis on the peri-oestrous period

Interrelationships of circulating hormone levels and their implications for follicular development were studied throughout the oestrous cycle with emphasis on the perioestrous period in heifers and cows. The oestradiol level showed a major peak (45 pmol/1) before and coinciding with oestrus, and a second peak (27 pmol/1) around day 5–6 (day 0: day of first standing oestrus); it was low during the luteal phase of the cycle when progesterone was higher than 14 nmol/1 from day −12 to day −2. Large antral follicles, which had developed during the luteal phase, did not secrete significant amounts of oestradiol, degenerated after luteolysis, and were replaced by a newly developing follicle which became preovulatory. Parallel with this development the oestradiol level increased from the onset of luteolysis to reach a plateau about 26 h before the onset of oestrus. The interval between the onset of luteolysis and the onset of oestrus was 58 h; luteolysis proceeded at a slower rate in heifers than in cows. At 4.6 h after the onset of oestrus the maximum of the LH surge was recorded; the LH surge appeared to be postponed in the period October–December in comparison to the period August–September. The maximum of the LH surge was higher in heifers (45 μg/l) than in cows (30 μg/l), but its duration was similar (8.0 h). The oestradiol level decreased significantly from 6 h after the maximum of the LH surge, and standing oestrus (duration 18 h) was terminated almost at the same time as the return to basal values of oestradiol. Cortisol and prolactin levels did not show a peak during the peri-oestrus period. Cortisol fluctuated irrespective of the stage of the oestrus cycle and prolactin was significantly higher during the luteal phase.

The results of this study indicate that development of the preovulatory follicle starts in the cow at the onset of luteolysis, about 2.5 days before the preovulatory LH surge, and that oestradiol secretion by this follicle is possibly inhibited by the LH surge.     

The effect of intrauterine and cervical manipulation on the equine oestrous cycle and hormone profiles.

Endometrial biopsy or endometrial biopsy and uterine culture taken on Day 4 after oestrus induced lysis of the corpus luteum (CL), resulting in a sharp decline in serum progesterone concentration and shortened the interoestrous interval in 8/12 and 32/33 oestrous cycles, respectively, during 2 experiments. Cervical dilatation 4 days after oestrus shortened the interoestrus interval in 5/10 and 0/5 oestrous cycles. Endometrial biopsy and culture on Days 1 and 3 after oestrus also induced CL lysis during 4 of 7 cycles. Total oestrogen (oestrone plus oestradiol) concentrations increased at the onset of the subsequent oestrus in mares biopsied on Day 4 of dioestrus or in control cycle oestrous periods. Endometrial biopsy also induced lysis of the CL in mares with persistent luteal function. It is postulated that intracervical or intrauterine manipulations during the luteal phase of the oestrous cycle may directly, or indirectly, stimulate the release of an endogenous luteolysin (prostaglandin) resulting in CL regression, followed by oestrus and ovulation in the mare.     

Luteolytic effect of 13,14-dihydro-PGF-2 alpha in heifers

Holstein heifers (4/group) were injected intramuscularly with 0, 5, 10 or 25 mg 13,14-dihydro-PGF-2 alpha on Day 10 of the oestrous cycle. Complete luteolysis and precocious oestrus occurred in 3 of 4 heifers receiving 25 mg and 1 of 4 receiving 10 mg 13,14-dihydro-PGF-2 alpha injected i.m. These features were not affected in heifers injected with 0 or 5 mg 13,14-dihydro-PGF-2 alpha, although plasma progesterone concentrations were depressed in all treated heifers within 75 min. LH concentrations were elevated between 5 and 8 h after 13,14-dihydro-PGF-2 alpha in all treated heifers. The addition of 13,14-dihydro-PGF-2 alpha to dispersed bovine luteal cells did not affect progesterone accumulation during a 2-h period. These results suggest that 13,14-dihydro-PGF-2 alpha may play a role in PGF-2 alpha-induced luteolysis.     

Uterine estrogen receptor alpha and progesterone receptor during the follicular and luteal phase in llamas

Estrogen receptor-alpha (ERalpha) and progesterone receptor (PR) were characterized in different endometrial cell types as luminal and glandular epithelium and stroma during the follicular (FP) and the luteal phase (LP) in llamas. Animals were examined daily by transrectal ultrasonography for the determination of the presence of an ovulatory follicle and ovulation was immediately induced by a GnRH injection (Day 0). Endometrial samples were obtained by transcervical biopsies from the left uterine horn on Day 0 (FP) and 9 days after the GnRH injection (Day 9, LP). Blood samples were collected on these days for estradiol 17beta and progesterone determination by RIA. An immunohistochemical technique was used to visualize ERalpha and PR immunostaining which was then analyzed by two independent observers. Total positive area and average staining for ERalpha were affected by the phase of the ovarian activity: in the three cell types there was more positive area and intense staining during the FP than during the LP. Similar findings were observed for PR, more positive stained areas were found during the FP than during the LP in the epithelia. In addition, the three cell types had more intense staining during the FP than during the LP. An effect of the cell type for ERalpha and PR was observed; epithelia (luminal and glandular) had more positive stained areas and greater intensity than stromal cells. In conclusion, the results of the present study suggest that in llamas, like in other ruminants, estradiol has a stimulatory effect while progesterone downregulates the ERalpha and PR and that the receptor is cell type specific.     

Growth and regression of ovarian follicles during the follicular phase of the oestrous cycle in heifers undergoing spontaneous and PGF-2 alpha-induced luteolysis

Ultrasonography was used to monitor the growth, ovulation and regression of individual ovarian follicles greater than or equal to 5 mm during the late luteal and follicular phases of the oestrous cycle in heifers treated with injections of PGF-2 alpha to induce luteolysis and in heifers undergoing spontaneous luteolysis. Six heifers were given a single injection of PGF-2 alpha between Day 12 and 15 of the oestrous cycle and their ovaries were examined daily by transrectal ultrasonography until ovulation occurred. Another group of 5 heifers was examined daily by ultrasound from Day 14 or 15 of the cycle through spontaneous luteolysis and ovulation. Blood samples were taken twice daily from this group and analysed for progesterone to determine when luteolysis occurred. All heifers were checked for oestrous behaviour twice daily. Mean diameters of ovulatory follicles on each of the 3 days before oestrus were not different between PGF-2 alpha-treated and untreated heifers. In both groups there was large variation among heifers in the sizes and growth rates of the ovulatory follicles. At 3 days before oestrus the diameters of ovulatory follicles were between 7.5 and 11 mm in PGF-2 alpha-treated heifers and between 6 and 11.5 mm in untreated heifers. Non-ovulatory follicles decreased in size during the 3 days before oestrus and the number of non-ovulatory follicles within the size ranges of ovulatory follicles decreased. The ovulatory follicle was not consistently the largest follicle on the ovaries until the day of oestrus but was always one of the 2 largest follicles during the 3 days before oestrus.(ABSTRACT TRUNCATED AT 250 WORDS)     

Progesterone profile in buffaloes during various stages of oestrous cycle using radioimmunoassy technique

Investigation were carried out to study the norms of progesterone concentration in the blood serum of buffaloes during various phases of oestrous cycle. Twenty four animals (12 heifers and 12 cows) were used. The blood serum samples were stored at -20 degrees C until processed for progesterone assay. The progesterone concentrations were measured by the radioimmunoassay technique. The progesterone levels were 0.360 +/- 0.062 and 0.334 +/- 0.066 ng/ml on the day of oestrus in buffalo-heifers and buffalo-cows, respectively. The values were around 1 ng/ml till day 6, followed by a gradual increase to a peak average value of 4.888 +/- 0.399 and 5.119 +/- 0.415 ng/ml on day 15 of the cycle in heifers and cows, respectively. Thereafter, the progesterone concentration fell abruptly to a level similar to that at oestrus. The mean progesterone value a day before oestrus was 0.488 +/- 0.067 and 0.577 +/- 0.053 ng/ml in buffalo-heifers and buffalo-cows, respectively. The mean progesterone concentration of different days of the cycle (except day 16) did not differ significantly (P / -0.01) between heifers and cows.     

Synergistic effects of insulin-like growth factor I and gonadotrophins on relaxin and progesterone secretion by ageing corpora lutea of pigs.

Insulin-like growth factor I (IGF-I) is involved in paracrine/autocrine regulation of gonadal steroidogenesis and peptide hormone biosynthesis. This study was designed to determine whether IGF-I alone, or an interaction of IGF-I, is involved in augmenting the actions of luteinizing hormone (LH) and prolactin in controlling relaxin and progesterone secretion from ageing corpora lutea of hysterectomized gilts at days 110, 113 and 116 after oestrus. Luteal tissue slices were incubated for 8 h with IGF-I (0, 50, 300 ng ml-1), LH (0, 100, 1000 ng ml-1), and prolactin (0, 100, 1000 ng ml-1) alone or in combination. Progesterone and relaxin concentrations were determined by radioimmunoassay of spent medium and of homogenates from luteal tissue slices before and after incubation. Porcine luteal tissue from day 110 had a net output of 25 ng progesterone and 26 ng relaxin in the control and of 65 ng progesterone and 2125 ng relaxin in the combined IGF-I, LH and prolactin treatment mg-1 of luteal tissue, respectively. IGF-I, LH and prolactin alone or in combination significantly increased (P < 0.01) progesterone production by luteal tissue from day 110, but they were partially effective at day 113 and ineffective at day 116. By contrast, the same hormone treatments increased relaxin production by luteal tissue from days 110 and 113. Even at day 116, prolactin alone or with LH or IGF-I continued to stimulate relaxin production. In conclusion, IGF-I augments the ability of prolactin and LH to increase relaxin production by ageing corpora lutea; however, a decrease in progesterone secretion and an increase in relaxin secretion at day 113 indicate that different mechanisms control progesterone and relaxin secretion in pigs.     

Control of oestrus in cattle using progesterone coils

Four experiments were conducted to determine the effect of length of treatment, stage of cycle at start of treatment and administration of oestradiol benzoate or progesterone at the start of treatment with intravaginal progesterone coils on oestrous response and fertility. In Experiment 1, the number of heifers in oestrus was affected neither by injection of 5 mg oestradiol benzoate alone or with 200 mg progesterone nor by length of treatment. More heifers (P < 0.05) were in oestrus on day 2 after treatment following a 12-day treatment compared to a 9-day treatment.In Experiment 2, heifers between days 17 and 20 of the oestrous cycle received an injection of either 5 mg oestradiol benzoate alone or with 200 mg progesterone at the start of a 9-day treatment with progesterone coils. Neither the number of heifers in oestrus nor the pattern of onset were affected after treatment. In Experiment 3, heifers between days 0 and 3 of the oestrus cycle received progesterone coils for 9, 12 or 14 days. In addition, animals received (i) no further treatment, (ii) a gelatin capsule adhered to the coil containing 10 mg oestradiol benzoate (iii) a gelatin capsule adhered to the coil containing 200 mg progesterone. Following a 9- or 12-day treatment period heifers receiving the coil with the oestrogen capsule had a high oestrous response ( compared to , P < 0.05). When oestrogen was not given, there was a significant linear effect of duration of treatment on the number in oestrus (9 days, ; 12 days ; 14 days, ; P < 0.05).In Experiment 4, post-partum cows were used to compare a 9- and 12-day treatment period and half the animals in each group received either 5 mg oestradiol benzoate and 200 mg progesterone at the start of treatment or a 10 mg gelatin capsule adhered to the coil. The length of treatment affected the number of heifers in oestrus since were in oestrus after a 12-day treatment period compared with after a 9-day period (P < 0.001). There were no significant differences in the number of cows in oestrus after injection of oestrogen and progesterone ( ) or after the use of the gelatin capsule ( ).     

Messenger RNA levels of estrogen receptors alpha and beta and progesterone receptors in the cyclic and inseminated/early pregnant sow uterus

The aim of the present study was to investigate differences in the expression of mRNAs for ERalpha, ERbeta and PR in the sow uterus at different stages of the estrous cycle as well as in inseminated sows at estrus and during early pregnancy by use of solution hybridization and in relation to plasma levels of estradiol and progesterone. Uterine samples were collected at different stages of the estrous cycle and after insemination/early pregnancy. In the endometrium, the expression of ERalpha mRNA and PR mRNA was similar for cyclic and early pregnant groups. Both were highest at early diestrus/70 h after ovulation and ERalpha mRNA was lowest at late diestrus/d 19 while PR mRNA was lowest at diestrus and late diestrus/d 11 and d 19. The expression of endometrial ERbeta was constantly low during the estrous cycle but higher expression was found in inseminated/early pregnant sows at estrus and 70 h after ovulation. In the myometrium, high expression of ERalpha mRNA and PR mRNA was observed at proestrus and estrus in cyclic sows and at estrus in newly inseminated sows. Higher expression of myometrial ERbeta mRNA was found in inseminated/early pregnant sows compared with cyclic sows, although significant only at estrus. In conclusion, the expression of mRNAs for ERalpha, ERbeta and PR in the sow uterus differed between endometrium and myometrium as well as with stages of the estrous cycle and early pregnancy. In addition to plasma steroid levels, the differences between cyclic and inseminated/early pregnant sows suggest that other factors, e.g. insemination and/or the presence of embryos, influence the expression of these steroid receptor mRNAs in the sow uterus.     

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

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

Nuclear estrogen and progesterone receptors in the oviduct of heifers under natural and superovulated estrous cycles

Oviducts from 22 crossbred heifers were examined for the presence of nuclear estrogen (ERalpha) and progesterone (PR) receptors at different phases (estrus, metaestrus and diestrus) of naturally occurring estrous cycles and estrous cycles during which superovulation was induced. Receptors were detected by immunohistochemistry in the epithelial cells, connective tissue and muscular layer of oviductal infundibulum, ampulla, ampullary/isthmic transition and isthmus. Epithelial ERalpha was found along the entire oviduct regardless of the cycle phase and of the circulating concentrations of 17beta-estradiol (E(2)) and progesterone (P(4)). Epithelial PR was found mainly at the ampullary-isthmic transition and isthmus and more intensely at the estrus phase but their amount was not correlated with P(4) concentrations. ERalpha in the connective tissue was more abundant at the infundibulum and ampulla, regardless of the phase of the estrous cycle and of E(2) and P(4) circulating concentrations. PR in the connective tissue was found mostly at the ampulla, regardless of the estrous cycle phase but no correlations were found between amount and P(4) concentrations. ERalpha staining intensity in the muscular layer was similar at all phases of the estrous cycle and at all anatomical segments of the oviducts. However, PR staining was more intense at the isthmus during the metaestrus phase and it was negatively correlated with P(4) concentrations. In general, data from the present research suggest that P(4) exerts an inhibitory role upon ERalpha and PR. Also, no differences were found between animals subjected or not to superovulation.