Follicular development,oocyte viability and recovery in relation to follicular steroids,prolactin and glycosaminoglycans throughout the estrous period in superovulated heifers with a normal LH surge,no detectable LH surge,and progestin inhibition of LH surge

Estrous cycles of heifers (n = 137) were synchronized with prostaglandin (PGF_2α) and follicular development stimulated with follicle stimulating hormone. Twenty-eight animals were administered Norgestomet implants 12 hr prior to the initial PGF2α injection to suppress the LH surge that initiates ovulation. Animals were ovariectomized every 12 hr after the initial PGF2α (7–9/time, 12–108 hr and at 192 and 240 hr post PGF2α) and divided into three treatment groups to consist of: 1) animals exhibiting a normal luteinizing hormone (LH) surge (n = 86), 2) animals in which no LH surge was detected (n = 23), and 3) suppression of the LH surge via Norgestomet implants (72–108 hr, n = 28). Follicular diameter was measured and follicular fluid was collected for analysis of prolactin, estradiol, progesterone and glycosaminoglycan concentrations. Progesterone concentrations were increased in animals exhibiting an LH surge as compared to animals in which no LH surge was detected; primarily in large follicles (> 8 mm diameter) after the LH surge. Animals not exhibiting an LH surge also had increased follicular progesterone concentrations compared to Norgestomet-implanted animals (242.3 ± 36.3 vs 86.7 ± 6.4 ng/ml, respectively, P < .01), indicating some LH stimulation. Follicular estradiol in animals exhibiting an LH surge increased up to the time of LH surge detection and then declined whereas animals with no LH surge detected had follicular estradiol concentrations that declined after the PGF_2α injection. No differences were noted between those that did not exhibit an LH surge or in which the LH surge was suppressed with Norgestomet in relation to follicular estradiol concentrations. Follicular estradiol concentrations increased with follicular size in all treatment groups (P < .01). Follicular concentrations of prolactin were increased in small follicles (P < .05; ≤ 4 mm diameter) and follicular prolactin increased from 12 to 36 hr post PGF2α injection, then declined after the LH surge. Follicular glycosaminoglycan concentrations decreased with increases in follicular size (P < .01) and were higher in animals that did not exhibit an LH surge (P < .01). No differences in follicular glycosaminoglycans were noted between Norgestomet-implanted animals and those not exhibiting an LH surge. In the animals representing days 4 and 6 of the subsequent estrous cycle (192 and 240 hr post PGF2α), numbers of small-sized follicles were increased. Follicular progesterone and estradiol concentrations were related to atretic large follicles unovulated from the prior estrus and a wave of growth in small and medium follicles. Follicular prolactin and glycosaminoglycans increased with time of the new estrous cycle and were increased in smaller follicles (P < .01). Suppression of LH with progestin implants (Norgestomet) may relate to early effects of progesterone, which may not be totally eliminated at target tissues and subsequently alters the LH surge, steroidogenesis of the follicle, and ovulation. Oocytes were predominantly found in the follicular fluid from animals in which an LH surge was detected and in the buffer wash of follicles in which no LH surge was detected. Oocyte viability was higher in animals exhibiting an LH surge (75% viable) whereas the oocytes of Norgestomet-implanted animals were 75% degenerate.     

Timing of preovulatory endocrine events, estrus and ovulation in Brahman x Hereford females synchronized with norgestomet and estradiol valerate

The purpose of these studies was to investigate the pattern and timing of preovulatory endocrine events, estrus and ovulation in Brahman X Hereford (F1) heifers synchronized with norgestomet and estradiol valerate. In Exp. 1, 66 nulliparous and 191 primiparous Brahman X Hereford (F1) heifers were used to estimate the interval from norgestomet implant removal to onset of estrus. The mean interval from implant removal to onset of estrus was 29.8 +/- .5 h, with 80.9% exhibiting estrus within 48 h. Endocrine and reproductive characteristics were examined in detail during Exp. 2 with 37 primiparous heifers. Continuous observation for estrus, 6-h or 2-h blood sampling and ovarian palpation per rectum were employed. All animals were artificially inseminated 48 h after implant removal. Mean interval from implant removal to onset of estrus and to onset of the luteinizing hormone (LH) surge were closely related (r = .91; P less than .0001). Mean intervals from implant removal to ovulation, onset of estrus to ovulation and onset of LH surge to ovulation were 59.1 +/- 2.5 h, 23.3 +/- 1.4 h and 23.1 +/- 1.6 h, respectively. Approximately 73% of heifers exhibited estrus within 54 h after implant removal (optimal timing); conception rate was 59.3% in this subgroup. Conception rate of heifers that did not exhibit estrus within 54 h after implant removal or exhibited an LH surge later than 12 h after estrus (delayed timing) was 10%. Assessment of plasma estradiol-17 beta concentrations suggested that retarded selection and(or) maturation of the preovulatory follicle following implant removal delayed estrus and lowered conception in up to 28% of females timed-inseminated at 48 h.     

Cumulus and oocyte maturation and in vitro and in vivo fertilization of oocytes in relation to follicular steroids,prolactin, and glycosaminoglycans throughout the estrous period in superovulated heifers with a normal LH surge,no detectable LH surge,and progestin inhibition of LH surge

Crossbred heifers (n = 103) were synchronized to estrus with prostaglandin (PGF_2α) and superovulated with follicle stimulating hormone (FSH-P). Animals were ovariectomized every 12 hr after the PGF_2α injection (n = 7 to 9/time) up to 108 hr to monitor the follicular, hormonal, and oocyte changes associated with follicular development and ovulation. Twenty-eight animals were implanted with Norgestomet implants 12 hr before PGF_2α and ovariectomized at 72, 84, 96, and 108 hr post PGF_2α injection to monitor effects of progesterone and suppression of the luteinizing hormone (LH) surge on oocyte maturation and quality. Follicular fluid was collected and analyzed for progesterone, estradiol, prolactin, and glycosaminoglycan content in conjunction with cumulus maturation and nuclear stage of oocyte maturation. Analysis of in vivo matured oocytes by in vitro fertilization was carried out at 60, 72, 84, and 96 hr post PGF_2α and in vitro matured oocytes at 12 to 108 hr post PGF_2α. No developmental changes in cumulus cells surrounding the oocyte of small follicles was noted (≤ 4 mm dia) indicating a static population. Medium (> 4 ≤ 8 mm) and large size (> 8 mm) follicles developed to the corona radiata and loose cumulus stages in animals in which an LH surge was detected but cumulus status remained primarily in the tight cumulus stage for animals without an LH surge. The estradiol-to-progesterone ratio for tight cumulus (TC), corona radiata (CR), and loose cumulus (LC) stages was 1.8 ± .1, 1.0 ± .1, and .4 ± .2, respectively (P < .01). Nuclear maturation of oocytes in small follicles from animals without a detectable LH surge seem to indicate early maturation (48 to 72 hr post PGF_2α) in conjunction with a high percent of degenerate oocytes not seen in animals exhibiting an LH surge. Oocytes from medium size follicles matured to germinal vesicle breakdown (GVBD) and early meiosis (metaphase I; MI) stages of development in all treatments. Most oocytes were degenerate in Norgestomet-implanted animals. Oocytes from large follicles (> 8 mm dia) from animals exhibiting an LH surge were in MI and metaphase II (MII) stages (48 to 84 hr post PGF_2α) in preparation of ovulation whereas oocytes from animals not exhibiting an LH surge had oocytes that early matured to MII (48 to 72 hr post PGF_2α), later regressing to degenerate oocytes (84 to 108 hr). Follicular progesterone, estradiol, and prolactin increased with oocyte maturation, particularly in medium and large follicles. In vivo matured oocytes for fertilization (60, 72, 84, and 96 hr post PGF_2α) were nude (from the oviduct) and primarily CR from follicles. Tubal oocytes (37%) were fertilized more frequently by a single sperm than follicular oocytes (14.3%; P < .01) and single sperm penetration peaked at 72 hr post PGF_2α. Follicular hormone concentrations were not related to sperm penetration. Oocytes (n = 101) matured in vivo had lower fertilization potential from ovaries producing < 14 or > 50 follicles (39.3%) as compared to 21 to 45 aspirated follicles (68.2%; P < .05), with a peak penetration at 32 follicles (86.7% penetration). No treatment differences (LH surge or no detectable LH surge) were noted in relation to in vivo matured oocytes. Oocytes with single sperm penetration had the lowest estradiol/progesterone ratio of 2.2 vs polyspermic penetration of 13.7.     

Lack of LH response to exogenous estradiol in heifers with ACTH-induced ovarian follicular cysts.

The aim of the present study was to examine the LH response to exogenous estradiol in 4 heifers with ACTH-induced ovarian follicular cysts. During the control experiment, administration of estradiol 24 hr after PGF2alpha in luteal phase heifers resulted in a LH response in all 4 heifers. The LH response was obtained between 16-20 hr after estradiol administration. The peak LH concentration (Mean +/- SEM; 5.1 +/- 0.8 ng/ml) during the control study was significantly different (P<0.05) from the concentration after cyst formation. None of the 4 heifers responded to estradiol after ovarian cyst formation. This result suggests that heifers with ACTH-induced ovarian follicular cysts may have a defective hypothalamio-pituitary response to exogenous estradiol similar to cows with spontaneous ovarian cysts.     

Endocrine pathogenesis of postweaning anestrus in swine: response of the persistently anestrous sow to hormonal stimuli

The endocrine function of the individual components of the hypothalamo-hypophyseal-ovarian axis of the postweaning anestrous sow was evaluated by monitoring the sow's response to exogenous estradiol, gonadotropin releasing hormone (GnRH), and gonadotropins. Sows (4 to 6/group) not returning to estrus by 42.8 +/- 3.1 days after weaning were assigned to 1 of the following treatments: 10 micrograms of estradiol benzoate (EB)/kg of body weight; 200 micrograms of GnRH, 1,000 IU of pregnant mare's serum gonadotropin (PMSG); 1,000 IU of human chorionic gonadotropin (HCG); or 4 ml of saline solution plus 2 ml of corn oil. A preovulatory-like surge of luteinizing hormone [(LH) greater than 12 hours in duration] was observed in all weaned sows treated with EB. All EB-treated sows exhibited estrus and ovulated but none conceived. Sows given GnRH had transiently increased (less than 3 hours) LH concentrations that were not associated with estrus or ovulation. Treatment with PMSG caused an increase in peripheral concentrations of 17 beta-estradiol that was followed by an LH surge, estrus, ovulation, and conception. Treatment with HCG caused an increase in circulating concentrations of 17 beta-estradiol that was accompanied by a surge of LH in some sows and ovulation in all sows. Not all sows treated with HCG exhibited estrous behavior, but conception occurred in 2 of 3 sows that were mated at estrus. None of the sows treated with saline plus corn oil had consistent changes in circulatory concentrations of 17 beta-estradiol or LH and none exhibited estrus or ovulated.(ABSTRACT TRUNCATED AT 250 WORDS)     

Blood LH level and induced ovulation after hCG and PMSG treatment in ovarian quiescent cattle

Ovarian quiescent cattle bearing follicle with palpable size were treated with single intramuscular injection of 750-6,000 IU of human chorionic gonadotrophin (hCG) in 13 cases and 1,000-2,000 IU of pregnant mare serum gonadotrophin (PMSG) in 5 cases. Changes of blood luteinizing hormone (LH) level, estrus and ovulation after the treatments were examined. After the hCG treatment LH level became slightly high from 0.2-0.6 ng/ml of pre-treatment to 0.3-1.9 ng/ml of post-treatment and maintained the level up to ovulation without the ovulatory LH surge. Ovulation was induced about 36 hr after the treatment in 12 cases. The ovulations were all silent ovulations. After the PMSG treatment LH level became slightly high from 0.6 ng/ml of pre-treatment to 1.3 ng/ml of post-treatment and the level lasted until the ovulatory LH surge. The ovulatory LH surge occurred about 39 hr after the PMSG treatment in 4 cases with a peak of about 32 ng/ml. Ovulation was induced about 74 hr after the treatment in all 5 cases. Four cases showed estrus but one in which the LH surge could not be confirmed did silent estrus preceding the induced ovulations. It was demonstrated that hCG induced ovulation without the LH surge but PMSG induced the ovulatory LH surge and the subsequent ovulation in ovarian quiescent cattle.     

Effects of melatonin and progesterone administered to ewes in spring and summer

Melatonin (MEL) was evaluated for effects on LH, prolactin (PRL) and fertility in spring (Exp. 1, 2) and summer (Exp. 3 to 5). In Exp. 1, 17 ovariectomized ewes bearing estradiol implants were fed 3 mg MEL or vehicle for 44 d beginning May 1. Melatonin decreased (P less than .001) PRL levels but had no effect on LH secretion and response to GnRH. In Exp. 2, 12 ewes each received a 40-d MEL ear implant or a sham implant on March 31. Progesterone-releasing pessaries (CIDR) were applied for 12 d and were withdrawn concomitant with ram joining on May 7. Neither treatment stimulated follicular development or induced estrus or ovulation. Exp. 3 and 4 were contemporary 2 x 2 factorial trials with 24 ewes at each of two locations. Melatonin implants were administered on June 29 and CIDR on July 22. The CIDR were removed and rams (Exp. 3, vasectomized; Exp. 4, fertile) were joined on August 3. Days from introduction of rams to estrus were reduced (P less than .05) by CIDR but not by MEL. All ewes lambed in Exp. 4, and days to estrus and conception were reduced (P less than .001) by CIDR but not by MEL. Exp. 5 was designed like Exp. 4 except that MEL implants were inserted June 20 and rams were joined August 8. Intervals from introduction of rams to estrus were reduced (P less than .01) by both MEL and CIDR treatments.(ABSTRACT TRUNCATED AT 250 WORDS)     

Characterization of ovine follicles destined to form subfunctional corpora lutea

A study was done to test whether ovulatory follicles destined to form subfunctional corpora lutea differed from normal ovulatory follicles in steroidogenic function. Twenty-five ewes were treated with prostaglandin F2 alpha on d 11 of the estrous cycle, then unilaterally ovariectomized before (n = 13) or after (n = 12) the surge of luteinizing hormone (LH) at the induced estrus to collect "control" follicles, which would have produced normal corpora lutea. In 15 ewes, the second ovary was removed 63 to 84 h later to collect "treated" follicles before (n = 7) or after (n = 8) the second expected surge of LH. Five ewes (control) were allowed to ovulate from the remaining ovary at first estrus and another five (treated) at the second estrus (3 to 4 d later). Treated ewes had lower serum progesterone than control ewes during the ensuing cycle (P less than .05). Treated follicles contained less estradiol in the theca (4.4 +/- .6 vs 10.0 +/- 2.5 ng; P less than .05), less androstenedione (.1 +/- .1 vs 1.0 +/- .2 ng) and estradiol (.5 +/- .1 vs 2.9 +/- 2.2 ng) in the granulosa (P less than .05) and less progesterone in the follicular fluid (.8 +/- .4 vs 3.3 +/- .8 ng; P less than .05) than control follicles, when removed before the surge of LH. Follicles removed after the surge of LH did not differ. In conclusion, ovulatory follicles with low steroidogenic function became corpora lutea that secreted lower-than-normal quantities of progesterone.     

Feed intake and serum GH, LH and cortisol in gilts after intracerebroventricular or intravenous injection of urocortin

In three experiments (Exp), ovariectomized gilts received intracerebroventricular (ICV; Exp 1 - with restraint, Exp 2 - without restraint) or intravenous (i.v.; Exp 3) injections of urocortin or saline to assess effects on feed intake and serum GH, LH, and cortisol. Following a 20-hr fast, feed was presented at 1 hr (Exp 1) or 30 min (Exp 2 and 3) after injection (time = 0 hr) of saline or 5 (U5) or 50 (U50) μg/pig (Exp 1 and 2) or 5 μg/kg BW (Exp 3) of urocortin. Blood samples were collected every 15 min from –2 to 6 hr relative to injection and hormone data pooled 2 hr before and hourly after treatment. Treatment with U50 decreased feed intake, relative to saline (treatment x time interaction; P < 0.05), when delivered ICV but not i.v. A treatment by time interaction was detected for GH (Exp 1, 2, 3) and LH (Exp 1 and 2; P < 0.01). Serum GH increased over time (relative to −2 hr; P < 0.05) following treatment with urocortin but not saline regardless of route of administration. Conversely, in Exp 1 (U5 and U50) and Exp 2 (U50), LH decreased relative to −2 hr with a delayed decrease during Exp 1. Serum cortisol was not affected by treatment in Exp 1, but increased following urocortin in Exp 2 and 3 (treatment by time interaction, P < 0.01). These data provide evidence that urocortin modulates GH and LH concentrations and suppresses feed intake in gilts via mechanisms which may be independent of cortisol and may depend upon dose and route of administration.     

Effects of sodium pentobarbital on release of gonadotropins in ewes immediately after ovariectomy and after treatment with gonadotropin-releas ing hormone

Two experiments were conducted with ewes 9 to 11 days after estrus to determine whether the secretion of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) are controlled differentially. In experiment 1, gonadotropin-releasing hormone (GnRH) was injected (100 (μg/ewe) at time = 0 min into ewes in four treatment groups. The treatment groups (9 ewes/group) were: 1) periodic iv sodium pentobarbital (NaPen) vehicle from 0 min; 2) periodic iv NaPen from 0 min; 3) vehicle iv for 120 min then iv NaPen from 120 min; 4) vehicle iv for 150 min then iv NaPen from 150 min. A surgical plane of anesthesia was maintained from the initiation of NaPen injection until the experiment ended. Jugular blood was sampled at 30-min intervals from ?30 to + 210 min for LH and FSH assays, and profiles of hormone concentrations were compared by time-trend analyses. GnRH released LH (P<.001) and FSH (P<.001), but NaPen did not affect the profiles of hormone concentrations; this indicated that NaPen did not reduce the ability of the pituitary to secrete gonadotropins in response to GnRH. Experiment 2 was a 2x2 factorial with ovariectomy (time = 0 hr) and NaPen as the main effects. One group of ovariectomized (n = 6) and one group of sham ovariectomized (n = 6) ewes were anesthetized only during surgery, while a group of ovariectomized (n = 7) and a group of sham ovariectomized (n = 6) ewes were kept at a surgical plane of anesthesia until 10 hr after surgery. Patterns of LH and FSH were compared in jugular blood collected hourly from 0 hr until 10 hr after surgery and in samples collected at 24 hr intervals from -24 to +72 hr of surgery. After ovariectomy, LH increased (P<.001) hourly and daily, but anesthesia suppressed (hourly, <.001 and daily, P<.005) these increases, which resulted in an interaction (hourly, P<.001 and daily, P<.01) of ovariectomy and anesthesia. FSH after ovariectomy increased hourly and daily (hourly, P<.02 and daily, P<.001), but the effect of anesthesia and interaction of ovariectomy and anesthesia were not significant. Because NaPen did not alter secretion of LH or FSH after exogenous GnRH in experiment 1 while it blocked the postovariectomy increase in LH but not FSH in experiment 2, we concluded that the postovariectomy increase in LH resulted from increased hypothalamic secretion of GnRH. The mechanisms responsible for the postovariectomy increase in FSH secretion are not identical to those for LH. The mechanisms that control the postovariectomy secretion of FSH might involve factors that are not suppressible by NaPen or, alternatively, the differences in LH and FSH release after ovariectomy might reflect the removal of ovarian factors that suppress FSH but not LH secretion in intact ewes.     

The effect of pulsatile gonadotropin-releasing hormone and estradiol administration on luteinizing hormone and follicle-stimulating hormone concentrations in pituitary stalk-sectioned ovariectomized pony mares

Hourly pulses of gonadotropin-releasing hormone (GnRH) or bi-daily injections of estradiol (E2) can increase luteinizing hormone (LH) secretion in ovariectomized, anestrous pony mares. However, the site (pituitary versus hypothalamus) of positive feedback of estradiol on gonadotropin secretion has not been described in mares. Thus, one of our objectives involved investigating the feedback of estradiol on the pituitary. The second objective consisted of determining if hourly pulses of GnRH could re-establish physiological LH and FSH concentrations after pituitary stalk-section (PSS), and the third objective was to describe the declining time trends of LH and FSH secretion after PSS. During summer months, ovariectomized pony mares were divided into three groups: Group 1 (control, n = 2), Group 2 (pulsatile GnRH (25 μg/hr), n = 3), and Group 3 (estradiol (5 mg/12 hr), n = 3). All mares were stalk-sectioned and treatment begun immediately after stalk-section. Blood samples were collected every 30 min for 8 h on the day before surgery (DO) and 5 d post surgery (D5) to facilitate the comparison of gonadotropin levels before and after pituitary stalk-section. Additionally, jugular blood samples were collected every 12 hr beginning the evening of surgery, allowing for evaluation of the gonadotropin secretory time trends over the 10 d of treatment. On Day 10, animals were euthanized to confirm pituitary stalk-section and to submit tissue for messenger RNA analysis (parallel study). Plasma samples were assayed for LH and FSH by RIA. Mean LH secretion decreased from Day 0 to Day 5 in Groups 1 and 3, whereas LH secretion tended (P < 0.08) to decrease in Group 2 mares. On Day 5, LH was higher (P < 0.01) in Group 2 (17.26 ± 3.68 ng/ml; LSMEANS ± SEM), than either Group 1 (2.65 ± 4.64 ng/ml) or group 3 (4.28 ± 3.68 ng/ml). Group 1 did not differ from Group 3 on Day 5 (P < 0.40). Similarly, mean FSH levels decreased in all groups after surgery, yet Group 2 mares had significantly (P < 0.001) higher FSH concentrations (17.66 ± 1.53 ng/ml) than Group 1 or Group 3 (8.34 ± 1.84 and 7.69 ± 1. 63 ng/ml, respectively). Regression analysis of bi-daily LH and FSH levels indicated that the time trends were not parallel. These findings indicate: 1) Pituitary stalk-section lowered LH and FSH to undetectable levels within 5 d after surgery, 2) pulsatile administration of GnRH (25 μg/hr) maintained LH and FSH secretion, although concentrations tended to be lower than on Day 0, and 3) E2 did not stimulate LH or FSH secretion.     

Ovarian follicular changes after weaning in sows.

Three experiments investigated ovarian follicular development in sows whose litters were weaned at 28 to 31 d of lactation. Unilateral ovariectomy near the time of weaning was used to assess early follicular characteristics and to identify those sows that would not return to estrus within 10 d after weaning. This allowed segregation of and exclusion from the study those sows that had a prolonged interval from weaning to first estrus. In Exp. 1, 82 and 72% of the large follicles that were marked at 48 or 72 h after weaning (10 sows per time point) were subsequently identified as corpora lutea. In Exp. 2, sows (seven to nine per time point) were unilaterally ovariectomized at 0, 6, 12, 18, 24, or 48 h after weaning, and follicular fluid was evaluated for changes in steroid concentrations. Progesterone concentrations in fluid from medium-sized (4 to 6 mm) follicles increased by 6 h after weaning and then declined through 24 h concomitant with increases in testosterone and estradiol. For Exp. 3, follicular fluid and granulosa cells from individual follicles were obtained from sows (seven to nine per time point) at 0, 6, and 24 h after weaning. In follicular fluid, insulin-like growth factor I (IGF-I) concentrations were not correlated (P greater than .05) with concentrations of progesterone, testosterone, or estradiol, or with granulosa cell production of estradiol during culture in androstenedione-supplemented medium.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of estradiol-17β, para-chlorophenylalanine and quipazine on tonic and LHRH-induced secretion of luteinizing hormone in ovariectomized ewes

Changes in metabolism of serotonin (5-HT) might mediate the reduced tonic luteinizing hormone (LH) and increased pituitary responsiveness to luteinizing hormone releasing hormone (LHRH) caused by estradiol-17β (estradiol). Two experiments were conducted to determine effects of estradiol, para-chlorophenylalanine (PCPA), an inhibitor of synthesis of 5-HT, and quipazine, an agonist of 5-HT, on tonic and LHRH-induced secretion of LH in ovariectomized ewes during the summer. Tonic levels of LH were reduced, the interval from LHRH to peak of the induced surge was longer and the magnitude of release of LH was greater in ovariectomized ewes treated with estradiol than in controls. Neither PCPA nor quipazine affected tonic secretion of LH. In ovariectomized ewes not receiving estradiol, PCPA and quipazine increased the magnitude of the LHRH-induced release of LH. However, PCPA reduced pituitary sensitivity to LHRH when administered concomitantly with estradiol; treatment with quipazine attenuated this effect of PCPA. The interval to the peak of the induced surge of LH was not affected by PCPA or quipazine in estradiol-treated or control ovariectomized ewes. Based on these results it appears that 5-HT mediates or is required for estradiol to increase pituitary responsiveness to LHRH.     

Effects of suckling and low doses of estradiol on luteinizing hormone secretion during the postpartum period in beef cows

An experiment was conducted to test if suckling acutely suppressed circulating levels of LH during the postpartum period in beef cows. In addition, the influence of exogenous administration of low concentrations of estradiol on LH secretion during the postpartum period was evaluated. Twelve mature cows were randomly assigned before parturition to one of three treatments. Four intact cows were used as controls (INT). Eight cows were ovariectomized within the first 7 days following parturition. Four of these cows received a silastic 17β-estradiol implant subcutaneously at the time of ovariectomy (OVX-E); the remaining four cows received no further treatment (OVX). All cows were allowed to nurse one calf for 30 min daily between 1200 and 1230 hours for the duration of the experiment. Blood samples were collected at 12 min intervals for 6 hr before and 6 hr after suckling on days 9, 30, 44 and 58 postpartum. Mean interval (mean ± SE) to the first increase in peripheral progesterone concentrations indicative of the onset of ovarian luteal activity was detected in INT cows 37 ± 4.9 days postpartum. An acute effect of suckling on LH secretion did not occur in INT and OVX cows but mean LH concentrations were reduced in OVX-E cows following suckling on days 44 and 58. Mean LH concentrations remained low in INT cows; whereas, in OVX and OVX-E cows LH concentrations increased linearly (P<0.05) as the interval from time of ovariectomy increased. Cows in the OVX-E group had a higher mean concentration of LH than cows in the OVX group at 30, 44 and 58 days postpartum (P<0.05). Frequency of LH pulses did not differ between cows in the OVX and OVX-E groups at any period. Data from this experiment support the concept that suckling is acting in a chronic fashion to inhibit LH secretion during the postpartum period. In the absence of ovaries, chronic administration of exogenous estradiol in low concentrations has a positive effect on secretion of LH in the postpartum cow.     

First luteal tissue in ewe lambs: influence on subsequent ovarian activity and response to hysterectomy

Two experiments were conducted in peripuberal ewe lambs to determine (a) the influence of the first luteal structure [most frequently a transient (i.e., 1 to 4 d) structure] on subsequent ovarian activity and (b) a role for the uterus in its demise. In Exp. 1, 21 lambs were assigned randomly on the day of the first rise in progesterone in the plasma to (1) sham-operation, (2) removal of the nonluteal ovary, (3) removal of the luteal ovary and (4) removal of the luteal ovary plus progesterone replacement (5 mg given three times 12 h apart, initiated at surgery). No effect of treatment on subsequent ovarian activity was observed. In Exp. 2, four of 14 lambs were assigned randomly to be hysterectomized before their first rise in progesterone. The remaining 10 lambs, (five each) were sham-operated or hysterectomized on the day of the first rise in progesterone. All hysterectomized lambs (N = 9) exhibited a rise in progesterone and maintained elevated concentrations of progesterone, whereas sham-operated lambs initiated estrous cycles. Oviducts and uteri collected from lambs hysterectomized on the day of the first rise in progesterone in Exp. 2 were flushed for presence of oocytes and none were found. Similarly, no retained oocytes were found in histological preparations of first luteal structures obtained from eight lambs ovariectomized in Exp. 1. Concentrations of luteinizing hormone (LH) and prolactin were determined in daily samples collected from 19 lambs in Exp. 1. Luteinizing hormone increased and became more variable as lambs matured, whereas prolactin decreased with no detectable change in variability. It is concluded that the transient luteal structure is not required for sexual maturation and that its lifespan is uterine dependent.     

Effects of enclomiphene on estradiol-induced changes in LH secretion in ewes

Experiments were conducted to characterize the ability of the antiestrogen enclomiphene (ENC) to block the effects of estradiol on secretion of LH in ovariectomized ewes. To determine whether ENC could block an estradiol-induced LH surge, ewes (n = 4/group) were administered 10 to 250 mg ENC followed 30 min later by 25 micrograms estradiol. Ten or 25 mg ENC suppressed the estradiol-induced LH surge in one of four ewes, whereas 100- or 250-mg doses suppressed the LH surge in three and four of four ewes, respectively. In ewes that received a single treatment of 100 mg ENC plus 25 micrograms estradiol, serum concentrations of LH remained below 1 ng/ml for 3 wk. Compared with untreated ewes, the number of pituitary GnRH receptors was elevated (P less than .05) at 12 d and 28 d, but pituitary content of LH had decreased (P less than .05) by 28 d in ewes treated with 100 mg ENC. To determine whether ENC could block the inhibitory effects of estradiol on serum concentrations of LH, ewes received injections of .03, .1, 1 or 10 mg ENC every 4 d. Half the ewes treated with each dose also received estradiol implants. Injection of .03, .1 or 1 mg ENC alone did not affect serum concentrations of LH, whereas the 10-mg dose decreased serum concentrations of LH below 1 ng/ml by wk 1 of treatment. No dose prevented the inhibition of serum concentrations of LH caused by estradiol implants. In ovariectomized ewes, ENC was antagonistic to estradiol; it prevented the positive effects of estradiol required to induce an LH surge.(ABSTRACT TRUNCATED AT 250 WORDS)     

Mechanistic target of rapamycin is activated in bovine granulosa cells after LH surge but is not essential for ovulation

The LH surge induces functional and morphological changes in granulosa cells. Mechanistic target of rapamycin (mTOR) is an integrator of signalling pathways in multiple cell types. We hypothesized that mTOR kinase activity integrates and modulates molecular pathways induced by LH in granulosa cells during the preovulatory period. Cows were ovariectomized and granulosa cells collected at 0, 3, 6, 12 and 24 hr after GnRH injection. While RHEB mRNA levels increased at 3 and 6 hr, returning to basal levels by 12 hr after GnRH treatment, RHOA mRNA levels increased at 6 hr and remained high thereafter. Western blot analyses revealed increased S6K phosphorylation at 3 and 6 hr after GnRH injection. Similarly, mRNA levels of ERK1/2, STAR and EGR‐1 were higher 3 hr after GnRH treatment. Rapamycin treatment inhibited mTOR activity and increased AKT activity, but did not alter ERK1/2 phosphorylation and EGR1 protein levels in cultured bovine granulosa cells. Rapamycin also inhibited LH‐induced increase in EREG mRNA abundance in granulosa cells in vitro. However, intrafollicular injection of rapamycin did not suppress ovulation. These findings suggest that mTOR is involved in the control of EREG expression in cattle, which may be triggered by LH surge stimulating RHEB and S6K activity.     

Androgen and estradiol effects on gonadotropin secretion and response to GnRH in ovariectomized pony mares

In Exp. 1, 16 long-term ovariectomized pony mares were used to determine the effects of treatment with estradiol benzoate (EB) and dihydrotestosterone (DHT) benzoate alone, and in combination, on secretion of follicle stimulating hormone (FSH) and luteinizing hormone (LH) in daily blood samples and after three consecutive injections of gonadotropin releasing hormone (GnRH). Administration of EB alone, or in combination with DHT, every other day for 11 d reduced (P less than .05) concentrations of FSH and increased (P less than .05) concentrations of LH in daily blood samples, and increased (P less than .05) the secretion of both gonadotropins after administration of GnRH. Treatment with DHT alone had no effect (P greater than .10) on LH or FSH concentrations in daily blood samples and no effect on the LH response to exogenous GnRH. There was no interaction (P greater than .10) between DHT and EB treatment for any hormonal characteristic. In Exp. 2, the control mares and mares treated with DHT in Exp. 1 were equally allotted to treatment with vehicle or testosterone propionate (TP) every other day for six injections, and then GnRH was administered as in Exp. 1. Treatment with TP had no effect (P greater than .10) on LH or FSH concentrations in daily blood samples but increased (P less than .05) the FSH response to exogenous GnRH, confirming our findings in previous experiments. It is concluded that the TP-induced stimulation of FSH secretion after exogenous GnRH in ovariectomized mares may involve estrogens produced from aromatization of the injected androgen.(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of the embryo on intrauterine migration in sheep.

Mechanisms of intrauterine migration were examined in 55 ewes. In the first experiment, corpora lutea were removed from unilaterally ovariectomized ewes on d 4 (d 0 = estrus) and pregnancy was maintained by giving exogenous progesterone. In Exp. 2, the reproductive tract was altered surgically such that embryos initially entered the uterine horn contralateral to the site of ovulation. In Exp. 3, ewes received beads of silastic polydimethylsiloxane that released either cholesterol or estradiol-17 beta in an attempt to mimic embryonic synthesis of estradiol. In the fourth experiment, unilaterally ovariectomized ewes were superovulated and spacing of embryos within the uterus was then examined. In all experiments, ewes were slaughtered on d 15 and recovery of embryos or beads from each uterine horn indicated that migration had occurred. All ewes in Exp. 1 and 2 that had two conceptuses experienced embryonic migration. Beads impregnated with estradiol migrated farther (P less than .01) than cholesterol-containing beads (27.6 +/- 4.3 vs 12.5 +/- 1.6 cm, respectively). In Exp. 4, only one conceptus had migrated into the contralateral horn in all ewes. These results demonstrated that 1) embryonic migration was not affected by local vs systemic exposure to progesterone, 2) embryos migrated into the unoccupied horn, regardless of the initial horn of entry, 3) estradiol may stimulate embryonic migration, and 4) conceptuses were not equally distributed between horns.