Duration of the breeding season and response to reproductive manipulation in five breeds of sheep under northern prairie conditions

The breeding season was 157, 154, <126, 210 and 217 days for Rambouillet, Columbia, Suffolk, Rambouillet x Finnish Landrace and Columbia x Finnish Landrace ewes respectively. Treatment of cyclic ewes with pregnant mare serum gonadotropin (PMSG) (500 IU), following a 12-day treatment with progestin-containing intravaginal sponges, did not affect fertility, but did decrease the time from sponge removal to estrus, (control 48.0 +/- 3.1 hr; PMSG 39.4 +/- 1.8 hr) to the preovulatory surge of LH (control 52.7 +/- 2.8 hr; PMSG 39.0 +/- 1.7 hr) and FSH (control 52.3 +/- 2.9 hr; PMSG 42.8 +/- 1.6 hr) and caused an elevation of serum LH levels prior to the preovulatory surge (control 1.25 +/- 0.18 ng/ml; PMSG 2.31 +/- 0.22 ng/ml). Exposure of the purebred ewes to 18 hours of daylight in January, decreasing by 30 minutes a week subsequently, counteracted the seasonal reduction in the number of ewes lambing following induced breeding under natural daylight in May. Prolificacy was greatest in crossbred ewes and their fertility was not affected by season. Gestation period was longer for fall-bred ewes and varied with breed.     

Inter-breed variation in the postpartum ewe response to continuous low dose infusion of GnRH

During the nonbreeding season the pituitary and ovarian responses to a subcutaneous GnRH infusion were investigated in acyclic, lactating Mule ewes which exhibit a deep seasonal anestrus and in Finn x Dorset ewes in which seasonal anestrus is ill-defined. Each breed received 10 d of progestagen priming before being subdivided into 3 groups. In Group L + G, 5 lactating ewes received GnRH (250 ng/h sc) for 96 h; in Group D + G, 5 dry ewes received GnRH (250 ng/h sc) for 96 h; in Group L, 5 lactating ewes received saline vehicle for 96 h. The infusions began when lactating and dry ewes were approximately 28 d and 120 d post partum, respectively. Blood samples were collected for LH, progesterone and estradiol analysis. Estrous behavior was monitored between Day -4 and Day +7. On Day +7 the reproductive tract was also examined. In the Mule ewes the mean plasma LH concentration increased (P < 0.05) following minipump insertion in each treatment group, although mean LH levels were greater (P < 0.05) in Group D + G, than in either Group L + G or Group L. Following the GnRH infusion, mean plasma estradiol levels increased (P < 0.05) in Group D + G but not in Group L + G. A preovulatory LH surge and subsequent ovulation occurred in 5 5 , 2 5 and 0 5 ewes from Group D + G, L + G and L, respectively, and estrus was recorded in 5 5 , 1 5 and 0 5 of these ewes, respectively. The LH surges began earlier (P < 0.05) (43.2 +/- 6.8 h vs 77.0 +/- 1.0 h) and the ovulation rate was greater (2.2 +/- 0.37 vs 1.00 +/- 0.00) in Group D + G than Group L + G. In the Finn x Dorset ewes mean LH concentrations increased (P < 0.05), to a similar level following minipump insertion in Groups D + G and L + G, but not Group L. The elevated LH levels were accompanied by increased (P < 0.05) plasma estradiol levels in Group D + G, but not in Group L + G. The GnRH infusion culminated in an LH surge and estrous behavior in 5 5 , 1 5 and 0 5 ewes from Groups D + G, L + D and L, respectively. The interval to the LH surge was similar between Group D + G (48.4 +/- 6.6 h) and Group L + G (46.0 h). Ovulation was evident in those ewes which exhibited an LH surge plus one additional ewe from Group L + G. The mean ovulation rate was greater in Group D + G (4.00 +/- 1.05) than in Group L + G (1.5 +/- 0.50). These data show that continuous GnRH infusion can consistently induce out of season breeding in the nonlactating Mule and Finn x Dorset ewe but can not break combined seasonal and lactational anestrous in these breeds. Further, between-breed differences are evident in the site along the hypothalamic-pituitary-ovarian axis at which reproduction is compromised in ewes at the same chronological stage post partum.     

Causes for decreased fertility in out-of-season mated ewes

The interval from onset of estrus to preovulatory luteinizing hormone (LH) release, conception and fertilization rates, and number of accessory spermatozoa per ovum at 48 hr postmating in untreated cyclic ewes and in progestogen-pregnant mare serum gonadotropin (PMSG) treated, anestrous ewes were compared in efforts to identify sources of lowered fertility for matings induced in anestrous ewes with exogenous hormones. Blood samples for LH determination were collected at 0, 2, 4, 6, 8, 10, 12, and 24 hr after the onset of estrus. Conception and fertilization failure rates were determined at 48 hr, 12 days, or parturition. The progestogen-PMSG treated ewes had a shorter interval from onset of estrus to preovulatory LH release, lower conception rates, and fewer accessory spermatozoa than cyclic ewes had. Conception failure, rather than embryonic mortality, was the major cause of reduced fertility for the out-of-season mated ewes and apparently resulted from insufficient viable spermatozoa in the oviducts to fertilize the ova.     

Thyroid hormones mediate steroid-independent seasonal changes in luteinizing hormone pulsatility in the ewe

Thyroid hormones permit the increase in response to estradiol negative feedback in ewes at the transition to anestrus. In this study, we tested whether the thyroid hormones are also required for steroid-independent seasonal changes in pulsatile LH secretion. In experiment 1, Suffolk ewes were ovariectomized and thyroidectomized (THX) or ovariectomized only (controls) in late November. LH pulse frequency and amplitude were measured for 4 h in December, April, May, June, and August. Pulse frequency was also measured in the presence of estradiol-containing implants during the breeding (December) and early anestrus (March) seasons. As expected, in the presence of estradiol, pulse frequency declined between December and March in control but not THX ewes. In the absence of estradiol, a seasonal decline in frequency and an increase in amplitude occurred in control ewes, concurrent with lengthening photoperiod. A similar trend was seen in THX ewes, but the seasonal changes were lower in magnitude and not significant. In experiment 2, the same protocol was used (pulse measurements in December, May, and June) with a larger THX group size (n = 7). Results were similar to those of experiment 1 for controls. In THX ewes, pulse frequency did not change over time and was significantly elevated relative to that of controls during the summer. Pulse amplitude in THX ewes tended to increase during summer and did not differ from pulse amplitudes in control ewes. These results demonstrate that thyroid hormones are required for steroid-independent cycles in LH pulse frequency; however, some seasonal changes in amplitude still occur in the absence of thyroid hormones. This finding contrasts with the changes in estradiol negative feedback at the transition to anestrus, which are entirely thyroid hormone dependent.     

Effect of melatonin on daily LH secretion in intact and ovariectomized ewes during the breeding season.

This study was conducted to find out whether daily LH secretion in ewes may be modulated by melatonin during the breeding season, when the secretion of both hormones is raised. Patterns of plasma LH were determined in luteal-phase ewes infused intracerebroventricularly (icv.) with Ringer-Locke solution (control) and with melatonin (100 microg/100 microl/h). Response in LH secretion to melatonin was also defined in ovariectomized (OVX) ewes without and after estradiol treatment (OVX+E2). Basal LH concentrations by themselves did not differ significantly before, during and after both control and melatonin infusions in intact, luteal-phase ewes. However, single significant (P<0.05) increases in LH concentration were noted during the early dark phase in the control and 1h after start of infusion in melatonin treated ewes. In both OVX and OVX+E2 ewes, melatonin decreased significantly (P<0.01, P<0.05, respectively) mean plasma LH concentrations as compared to the levels noted before the infusions. In OVX+E2 ewes, a single significant (P<0.05) increase in LH occurred 1h after start of melatonin treatment, similarly as in luteal-phase ewes. No significant differences in the frequencies of LH pulses before, during and after melatonin infusion were found in all treatments groups. In conclusion, melatonin may exert a modulatory effect on daily LH secretion in ewes during the breeding season, stimulating the release of this gonadotropin in the presence of estradiol feedback and inhibiting it during steroid deprivation. Thus, estradiol seems to be positively linked with the action of melatonin on reproductive activity in ewes.     

Effects of breed, ovarian steroids and season on the pulsatile secretion of LH in ovariectomized ewes

The effects of season and of oestradiol and progesterone on the tonic secretion of LH were studied in ovariectomized Merino and Suffolk ewes, two breeds which differ markedly in the seasonal pattern of their reproductive activity. In the absence of exogenous steroids, the frequency of LH pulses was lower and the amplitude of the pulses was higher in anoestrus than in the breeding season for Merino and Suffolk ewes 30 days after ovariectomy. In long-term (190 days) ovariectomized ewes, this seasonal change in LH secretion was observed in Suffolk ewes only. During seasonal anoestrus, treatment of ewes with subcutaneous oestradiol-17 beta implants (3, 6 or 12 mm in length) decreased the frequency of LH pulses in a dose-dependent manner, with Suffolk ewes being far more sensitive to the inhibitory effects of oestradiol than Merino ewes. The lowest dose of oestradiol (3 mm) had no effect on the secretion of LH in Merino ewes, but reduced secretion in Suffolk ewes. Treatment of ewes with the highest dose of oestradiol (12 mm) completely abolished LH pulses in Suffolk ewes, whereas infrequent pulses remained evident in Merino ewes. During the breeding season, oestradiol alone had no effect on the pulsatile release of LH in either breed, but in combination with progesterone there was a significant reduction in LH pulse frequency. Progesterone effectively decreased LH secretion in both breeds in both seasons. It was concluded that differences between breeds in the 'depth' of anoestrus could be related to differences in the sensitivity of the hypothalamus to both negative feedback by oestradiol and the direct effects of photoperiod.     

Estrogen and leptin have differential effects on FSH and LH release in female rats

Prior experiments have shown that the adipocyte hormone leptin can advance puberty in mice. We hypothesized that it would also stimulate gonadotrophin secretion in adults. Since the secretion of follicle stimulating hormone (FSH) and luteinizing hormone (LH) is drastically affected by estrogen, we hypothesized that leptin might have different actions dependent on the dose of estrogen. Consequently in these experiments, we tested the effect of injection of leptin into the third cerebral ventricle of ovariectomized animals injected with either the oil diluent, 10 microg or 50 microg of estradiol benzoate 72 hr prior to the experiment. The animals were ovariectomized 3-4 weeks prior to implantation of a cannula into the third ventricle 1 week before the experiments. The day after implantation of an external jugular catheter, blood samples (0. 3 ml) were collected just before and every 10 min for 2 hr after 3V injection of 5 microl of diluent or 10 microg of leptin. Both doses of estradiol benzoate equally decreased plasma LH concentrations and pulse amplitude, but there was a graded decrease in pulse frequency. In contrast, only the 50-microg dose of estradiol benzoate significantly decreased mean plasma FSH concentrations without significantly changing other parameters of FSH release. The number of LH pulses alone and pulses of both hormones together decreased as the dose of estrogen was increased, whereas the number of pulses of FSH alone significantly increased with the higher dose of estradiol benzoate, demonstrating differential control of LH and FSH secretion by estrogen, consistent with alterations in release of luteinizing hormone releasing hormone (LHRH) and the putative FSH-releasing factor (FSHRF), respectively. The effects of intraventricularly injected leptin were drastically altered by increasing doses of estradiol benzoate. There was no significant effect of intraventricular injection of leptin (10 microg) on the various parameters of either FSH or LH secretion in ovariectomized, oil-injected rats, whereas in those injected with 10 microg of estradiol benzoate there was an increase in the first hr in mean plasma concentration, area under the curve, pulse amplitude, and maximum increase of LH above the starting value (Deltamax) on comparison with the results in the diluent-injected animals in which there was no alteration of these parameters during the 2 hr following injection. The pattern of FSH release was opposite to that of LH and had a different time-course. In the diluent-injected animals, probably because of the stress of injection and frequent blood sampling, there was an initial significant decline in plasma FSH at 20 min after injection, followed by a progressive increase with a significant elevation above the control values at 110 and 120 min. In the leptin-injected animals, mean plasma FSH was nearly constant during the entire experiment, coupled with a significant decrease below values in diluent-injected rats, beginning at 30 min after injection and progressing to a maximal difference at 120 min. Area under the curve, pulse amplitude, and Deltamax of FSH was also decreased in the second hour compared to values in diluent-injected rats. In contrast to the stimulatory effects of intraventricular injection of leptin on pulsatile LH release manifest during the first hour after injection, there was a diametrically opposite, delayed significant decrease in pulsatile FSH release. This differential effect of leptin on FSH and LH release was consistent with differential effects of leptin on LHRH and FSHRF release. Finally, the higher dose of E2 (50 microg) suppressed release of both FSH and LH, but there was little effect of leptin under these conditions, the only effect being a slight (P < 0.04) increase in pulse amplitude of LH in this group of rats. The results indicate that the central effects of leptin on gonadotropin release are strongly dependent on plasma estradiol levels. These effects are consistent w     

Ovarian antral follicular dynamics and their associations with peripheral concentrations of gonadotropins and ovarian steroids in anoestrous Finnish Landrace ewes

Daily transrectal ultrasonography of ovaries was done in seven Finn ewes during three 17-day periods from May to July. Blood samples were collected each day for estimation of the serum follicle-stimulating hormone (FSH), oestradiol and progesterone concentrations, and also every 15 min for 6 h, halfway through each period of ultrasonographic examination, to determine the patterns of gonadotropic hormone secretion. Four ewes ceased cycling from March to mid-April (ewes entering anoestrus early) and three in May (ewes entering anoestrus late). In all ewes cyclicity resumed during the period from mid-August to mid-September. The growth of ovarian antral follicles to periovulatory sizes of >/=5 mm in diameter was seen at all stages of anoestrus. An average of four waves of follicular development (follicles growing from 3 to >/=5 mm in diameter before regression) with a periodicity of 4 days were recorded during each of the three scanning periods. There was a close temporal relationship between days of follicular wave emergence and peaks of successive FSH fluctuations. Ewes entering anoestrus late exceeded ewes that became anoestrus early in numbers of large (>/=5 mm in diameter) ovarian antral follicles and maximum follicle diameter. Peak concentrations of transient FSH increases were higher (P<0.05) in ewes entering anoestrus late than in ewes entering anoestrus early. The secretion of luteinising hormone, (LH; mean and basal level, and LH pulse frequency, but not amplitude) was lowest during the month of June in all ewes. Oestradiol production was markedly suppressed throughout anoestrus. Peaks of progesterone secretion appeared to occur at regular intervals and were associated with the end of the growth phase of the largest follicles of sequential waves. In conclusion, the growth of ovarian follicles to ostensibly ovulatory diameters is maintained throughout anoestrus in Finn ewes and periodic emergence of follicular waves is correlated with an endogenous rhythm of FSH secretion. The present study also provides evidence for the inverse relationship between the time of the onset of seasonal anoestrus and the number and size of antral follicles developing throughout anoestrus in Finn ewes, and indicates that differences exist in both the secretion of and ovarian responsiveness to gonadotropic hormones among early and late anoestrous ewes.     

Regulation by estradiol of hypothalamic somatostatin gene expression: possible involvement of somatostatin in the control of luteinizing hormone secretion in the ewe

In the ewe, the mediobasal hypothalamus (MBH) is the primary central site for estradiol to generate the preovulatory GnRH/LH surges and sexual behavior. This area contains numerous neurons expressing the estradiol receptor alpha, distributed in the ventromedial nucleus (VMN) and the infundibular nucleus (IN). A large proportion of these neurons express somatostatin, making this neuropeptide a potential candidate for transmission of the estradiol signal to the GnRH neurons located in the preoptic area. We tested this hypothesis using ovariectomized ewes that had been subjected to an artificial estrous cycle. In the first experiment, 22 h after progesterone removal, ewes received estradiol (treated ewes) or empty implants (control ewes) for 4 h and then were killed. Using in situ hybridization, we showed that this short estradiol treatment increased the somatostatin mRNA amount by about 50% in the VMN and 42% in the IN. In the second experiment, preovulatory estradiol signal was replaced by somatostatin intracerebroventricular (ICV) administration. This treatment abolished LH pulsatility and dramatically decreased the mean basal level of LH secretion while it did not affect the mean plasma GH concentration. We demonstrated that an increase in somatostatin mRNA occurs at the time of the negative feedback effect of estradiol on LH secretion during the early stage of the GnRH surge induction. As ICV somatostatin administration inhibits the pulsatile LH secretion by acting on the central nervous system, we suggest that somatostatin synthesized in the MBH could be involved in the estradiol negative feedback before the onset of the preovulatory surge.     

Endocrine response of postpartum anestrous beef cows to GnRH or PMSG

Forty-one postpartum anestrous Hereford cows, maintained under range conditions, were used to determine the influence of gonadotropin releasing hormone (GnRH) or pregnant mare serum gonadotropin (PMSG) on ovarian function. Anestrous cows were identified by estrous detection with sterile bulls and concentrations of progesterone in plasma obtained weekly. At 45 +/- 2 days postpartum, cows were allotted to the following treatments: (1) control (saline), (2) 100 mug GnRH, (3) 200 mug GnRH, (4) 200 mug GnRH in carboxymethyl cellulose (CMC), (5) 500 IU PMSG, (6) 1,000 IU PMSG or (7) 2,000 IU PMSG. Cows were bled frequently the first day after treatment and then every other day until 85 days postpartum. The LH responses after 100 and 200 mug of GnRH were not significantly different and mixing 200 mug GnRH with CMC before injection did not significantly alter the LH response. During the first 20 days after treatment, neither GnRH nor 500 IU PMSG altered estradiol concentrations in plasma, but treatment of cows with 1,000 or 2,000 IU PMSG resulted in increased (P<0.01) concentrations of estradiol. The time postpartum required for concentrations of progesterone in plasma to exceed 1 ng/ml was reduced (P<0.05) by all treatments except 100 mug GnRH. These data indicate that GnRH causes LH release in anestrous range cows and that treatment with 1,000 or 2,000 IU PMSG initiates ovarian activity as evidenced by increased concentrations of estradiol in plasma.     

Effects of suckling mode on endocrine control of reproductive activity resumption in Texel ewes lambing in July or November

To study the effects of the overlapping of seasonal and lactation anestrus and the influence of the suckling mode on the resumption of reproductive activity in Texel ewes, two experiments were carried out after the July and November lambings. The frequency and the duration of suckling decreased with the age of the lambs, and the suckling intensity in the lambs that were allowed to suck unrestrictedly was three times higher than in lambs restricted to sucking three times a day. The overlapping of seasonal anestrus and lactation delayed the resumption of ovarian and estrus activity, but no difference was observed between dams suckling single lambs and twins. The resumption of ovarian and estrus activity was much earlier after the November lambing than after the July lambing. Weaning after the November lambing shortened the interval between parturition to first estrus but not to the first luteal function. The reduction of suckling intensity by suckling management had no effect on the resumption of ovarian and estrus activity. In early postpartum, suckling inhibited the luteinizing hormone (LH) pulsatile secretion and consequently the first LH discharge. However, the earlier restoration of gonadotropin stimulation in dry ewes was not immediately followed by ovarian activity. The suckling inhibition may be due to a temporary disturbance in hormonal balance, the rise in prolactin (PRL) and cortisol secretions. Plasma estradiol 17beta (E2) concentrations were higher in dry than in suckling ewes in early postpartum. Follicle stimulating hormone (FSH) secretion was not involved in the process of delaying the resumption of reproductive activity after lambing.     

Some effects of epidermal growth factor on reproductive function in Merino sheep

During the i.v. infusion of a depilatory dose (100 micrograms/kg bodyweight) of mouse epidermal growth factor (EGF) into ovariectomized Merino ewes the frequency of pulsatile LH release was significantly reduced. However, the amplitude of pulses of LH secretion, either those naturally occurring or those induced by LHRH injection, was unchanged or only slightly reduced. Similar infusions of mouse EGF were made in progestagen-treated anoestrous Merino ewes in which LH secretion was maintained by injections of LHRH. These ewes did not experience oestrus or ovulate in response to PMSG injected 1 day after mouse EGF treatment (2 days before progestagen withdrawal); both responses occurred in controls. The EGF-treated ewes experienced oestrus and ovulated following progestagen-PMSG treatment 6 weeks later. These results suggest that mouse EGF inhibits the hypothalamic pulse generator responsible for LH release in the ewe but has little if any effect on pituitary sensitivity to LHRH; and mouse EGF apparently has a direct effect on the ovaries, temporarily impairing their ability to ovulate in response to exogenous gonadotrophin.     

Evidence that estrogen receptor alpha,but not beta,mediates seasonal changes in the response of the ovine retrochiasmatic area to estradiol

In ewes, anestrus results from a reduction in LH pulsatility due to an increased sensitivity of the hypothalamic estradiol negative feedback system. Considerable evidence has implicated the A15 group of dopaminergic neurons in the retrochiasmatic area in this seasonally dependent estradiol effect. Moreover, estradiol administered to the retrochiasmatic area in ovariectomized anestrous ewes inhibits LH secretion. However, A15 neurons do not appear to contain the classical estrogen receptors (ERalpha). Therefore, we tested the hypothesis that beta-estrogen receptors mediate the action of estradiol in the retrochiasmatic area by comparing the effects of estradiol and genistein, a selective ERbeta agonist. We also examined whether there are seasonal changes in response of the retrochiasmatic area to these agonists and if these effects are mediated by dopamine. To test these hypotheses, ovariectomized ewes were implanted with bilateral guide cannulae targeting the retrochiasmatic area. Crystalline agonists were administered via microimplants inserted down the cannulae. Blood samples taken before and 4 days after microimplant insertion were analyzed for LH concentrations, pulse frequency, and amplitude. Genistein treatment produced no significant change in LH levels in either season. Estradiol treatment decreased both mean LH concentrations and pulse frequency in anestrous but not breeding-season ewes. Administration of the dopamine antagonist sulpiride to ovariectomized ewes with estradiol microimplants in the retrochiasmatic area returned LH pulse frequency to levels indistinguishable from controls. From these data, we hypothesize that estradiol acts on local ERalpha-containing neurons in this area to stimulate a dopaminergic pathway that inhibits LH secretion during anestrus.     

Progesterone directly inhibits pituitary luteinizing hormone secretion in an estradiol-dependent manner.

We recently demonstrated that progesterone and estradiol inhibit pituitary LH secretion in a synergistic fashion. This study examines the direct feedback of progesterone on the estradiol-primed pituitary. Nine ovariectomized (OVX) ewes underwent hypothalamic-pituitary disconnection (HPD) and were infused with 400 ng GnRH every 2 h throughout the experiment. After 7 days of infusion, estradiol was implanted s.c. Four days later, estradiol implants were exchanged for blank implants in 4 ewes and for progesterone implants in 5 ewes. These implants remained in place for another 4 days. Blood samples were collected around exogenous GnRH pulses before and 0.5 to 96 h after implant insertion and exchange. Serum LH and progesterone concentrations were determined through RIA. One month later, 4 of the HPD-OVX ewes previously implanted with steroids were reinfused with GnRH and the implantation protocol was repeated using blank implants only. In estradiol-primed ewes, progesterone significantly lowered LH secretion after 12 h of implantation and LH secretion remained inhibited while progesterone implants were in place (p less than 0.05). Removing estradiol transiently lowered LH secretion, and this effect was significant only 24 h after estradiol withdrawal (p less than 0.05). These data suggest that progesterone has a direct, estradiol-dependent inhibitory effect on pituitary LH release and that estradiol may sustain pituitary gonadotrope response to GnRH.     

Dynamic changes in the gonadotrope cell subpopulations during an estradiol-induced surge in the ewe

Whether estradiol targets a subpopulation of gonadotrope cells was investigated in this study. Ovariectomized ewes (OVX) or OVX ewes immunized against GnRH and treated with hourly pulses of GnRH analogue (OVX-IMG) were killed at 6, 12, 16, and 24 h after administration of 50 microg of 17beta-estradiol (E(2)). Control ewes received no E(2) treatment. In OVX or OVX-IMG ewes killed 6 h after E(2) injection, a decrease in gonadotropin plasma levels was observed compared with non-E(2)-treated ewes. In contrast, a surge in gonadotropin plasma concentrations occurred in ewes killed 16 h after injection. The percentage of total immunoreactive gonadotrope cells among the pituitary cells was lower in E(2)-treated ewes compared with nontreated animals. The proportion of monohormonal LH cells was constant throughout the experiment, except at the surge peak, where it was enhanced. In the OVX ewes, the proportion of bihormonal LH/FSH cells was lower in the E(2)-treated ewes compared to the nontreated ewes (P: < 0.001), with a more pronounced decrease 16 h after E(2) injection. A slight increase occurred 12 h after E(2) injection compared with 6 h after injection (P: < 0.05). A similar pattern was observed in the OVX-IMG ewes, except at 12 h after E(2) injection, when no increase occurred. In both OVX and OVX-IMG ewes, injection of E(2) decreased FSHbeta mRNA expression but did not alter the relative levels of LHbeta mRNA. These data suggest that the negative feedback of E(2) on LH and FSH secretion mainly targets the bihormonal cells and occurs, at least in part, directly at the pituitary level. During the gonadotropin surge, the sustained FSH release from the bihormonal cells would induce a switch from bihormonal cells to monohormonal LH cells by depleting these cells of FSH.     

Effect of rumen degradable bolus containing melatonin or progestagen pessary plus pregnant mare serum gonadotropin on estrus response and lambing rate in ewes

Two practical regimens designed to induce estrus and ovulation in ewes in late anestrus were compared. Forty ewes were given a soluble glass rumen bolus containing 150 mg melatonin on July 9 and were joined with two vasectomized rams on July 23 and with three fertile rams on August 6. A second group of 40 ewes was treated with an intravaginal progestagen pessary (60mg medroxy-progesterone acetate) on July 23. Following pessary removal after 12 d, ewes were given 750 IU of pregnant mare serum gonadotropin (PMSG). Five fertile rams were joined with these ewes 48 h after progestagen removal. Melatonin concentrations were determined in single blood samples collected in early afternoon of July 21. Mating dates, lambing dates and litter sizes were recorded. Date of mating was significantly later in ewes treated with melatonin compared with those treated with progestagen plus PMSG (P<0.0001). All ewes given melatonin were mated within 4 wk, and those on progestagen plus PMSG treatment within one day of fertile ram introduction. Thirty-four ewes (85%) allocated to melatonin treatment and 36 (90%) allocated to progestagen plus PMSG treatment lambed (P>0.05). Mean (+/-SEM) lambing date was later in melatonin-treated ewes (January 17+/-1.2 d) compared to those given progestagen plus PMSG (December 30+/-0.6 d; P<0.0001). Mean litter size was lower in melatonin-treated ewes (1.5+/-0.1) compared with those given progestagen plus PMSG (2.0+/-0.1; P<0.001). Plasma melatonin concentrations indicated that 9 of 40 ewes treated with melatonin had circulating melatonin concentrations of less than 16 pg/ml. It is concluded that under conditions that existed in this experiment, treatment with progestagen plus PMSG in late anestrus resulted in more synchronous mating and lambing and a higher litter size than that following administration of a soluble glass rumen-degradable bolus containing melatonin.     

Plasma luteinizing hormone and prolactin in normothermic and hyperthermic ovariectomized ewes

The effect of bromocriptine on concentrations of luteinizing hormone (LH) and prolactin (PRL) as well as the rhythmicity of episodic profiles of plasma LH were investigated in twelve ovariectomized ewes exposed to 3-day trials during which ambient temperature/humidity conditions maintained either normothermia or induced an average of 1.4°C increase of rectal temperature (hyperthermia). In 24 of 48 trials, ewes received twice daily subcutaneous injections of 1 mg bromocriptine beginning at 1900 hr on day 1. Plasma PRL and LH were measured at 10-min intervals for 4 hr on days 2 and 3. Bromocriptine significantly decreased plasma PRL (65 ± 6 vs 5 ± 1 ng/ml), mean plasma LH (11.0 ± 0.2 vs 6.5 ± 0.2 ng/ml) and tended (P < 0.1) to decrease LH rhythmicity. In hyperthermic placebo-treated ewes, plasma PRL was increased (65 ± 6 vs 212 ± 20 ng/ml) and mean LH was decreased (11.0 ± 0.2 vs 8.2 ± 0.2 vg/ml) compared to normothermic, placebo-treated ewes, but there was no effect of hyperthermia on LH rhythmicity. Bromocriptine treatment of hyperthermic ewes decreased mean PRL (212 ± 20 vs 32 ± 9 ng/ml) on both days of sampling although mean levels were significantly higher on day 2 than on day 3(54 ± 14 vs 10 ± 6 ng/ml). Perhaps because mean LH was already inhibited in hyperthermic ewes, bromocriptine did not further decrease mean LH (8.2 ± 0.2 vs 6.6 ± 0.2 ng/ml), but LH rhythmicity was decreased (P < 0.01). There was no significant difference in mean LH between normothermic ewes receiving bromocriptine and hyperthermic ewes receiving bromocriptine (6.5 ± 0.2 vs 6.6 ± 0.2 ng/ml). These results indicate that bromocriptine inhibits PRL and LH secretion in normothermic ewes. In hyperthermic ewes, the inhibitory effect of bromoriptine on PRL was even more pronounced, but the effect on LH release was minimal perhaps because LH was already inhibited by hyperthermia.     

Effects of progestagen treatments and PMSG on the induction of the preovulatory LH discharge in ewes

The characteristics of the induced preovulatory LH discharge were compared in ewes after treatment for 12 days with intravaginal sponge pessaries impregnated with 40 mg Fluorogestone Acetate or with subcutaneous ear implants containing varying quantities of Norgestomet. In Experiment 1, ewes were treated with intravaginal sponges or implants alone. In Experiment 2, ewes received similar treatments and 500 IU pregnant mares' serum gonadotropin (PMSG) i.m. at the time of sponge or implant removal. The duration of the LH discharge and an estimate of the total LH discharged were similar among treatment groups within the same experiment. Overall, the onset of LH release occurred approximately 8 h earlier in ewes treated with implants, whether or not PMSG was used. Use of PMSG, in conjunction with implant or sponge treatments, shortened the mean interval from sponge or implant removal to the onset of LH release from 41 to 28 h and doubled the estimated total LH discharged, compared with treatments using sponges or implants alone.     

Genetic and environmental variation in the LH response of ovariectomized sheep to LH-RH.

The influence of breed and season on the sensitivity of the pituitary gland of sheep to LH-RH was assessed. Ovariectomized ewes of 3 breeds (Finnish Landrace, Scottish Blackface and Tasmanian Merino) with differing normal breeding seasons and with differing ovulation rates were injected (i.v.) with 3 doses of LH-RH (1.56, 6.25 or 25.0 micrograms) at 3 different times of the year covering the anoestrous and the breeding seasons of intact ewes; 9 ewes of each breed (3 per sub-class) were examined on the first and third occasions, 6 (2 per sub-class) on the second. The response was measured in terms of the concentration of LH in peripheral plasma 20, 40, 60 and 80 min after injection. Time of year, but not the breed of sheep, affected the magnitude of the response; the data indicated that the duration of LH secretion was greater during the breeding season than during anoestrus. It was concluded that changes in the spontaneous activity of the hypothalamus/hypophysis could contribute to seasonal changes in LH secretion independently of the modifying effects of gonadal steroids. Such variation in unmodulated activity apparently does not contribute to the differences in ovulation rate among the 3 breeds.     

Role of estradiol in inducing an ovulatory-like surge of luteinizing hormone in sheep

Two experiments were performed to examine the effect of estradiol on secretion of luteinizing hormone (LH) and on the number of receptors for gonadotropin-releasing hormone (GnRH) after down regulation of GnRH receptors in ovariectomized ewes. In the first experiment, ovariectomized ewes were administered one of four treatments: Group 1) infusion of GnRH i.v. for 40 h; Group 2) injection of 100 micrograms estradiol i.m.; Group 3) infusion of GnRH i.v. for 16 h followed immediately by an injection of 100 micrograms estradiol i.m.; and Group 4) infusion of GnRH i.v. for 40 h plus injection of 100 micrograms estradiol i.m. after the 16th h of infusion. Ewes in Groups 1, 3 and 4 responded to the infusion of GnRH with an immediate increase in serum concentrations of LH, with maximum values occurring between 2 and 4 h after the start of infusion; serum concentrations of LH then began to decline and were approaching the pretreatment baseline within 16 h. Administration of estradiol resulted in a surge of LH regardless of whether the pituitary had been desensitized by infusion of GnRH or not. In all cases the magnitude of the surge was similar to that induced by the initial infusion of GnRH. In Groups 2 and 3 the surge of LH began at 12.3 +/- 0.1 and 11.9 +/- 0.1 h after administration of estradiol. In contrast, the ewes in Group 4 had a surge of LH beginning 3.7 +/- 0.1 h after administration of estradiol.(ABSTRACT TRUNCATED AT 250 WORDS)