Effect of calf removal on serum luteinizing hormone and cortisol concentrations in postpartum beef cows

Serum luteinizing hormone (LH) and cortisol concentrations were measured in ten fall calving, Angus cows averaging 38 +/- 8 days postpartum. Calves from five cows were weaned at the beginning of the study. Blood samples were collected at 20 min. intervals for 48 h after weaning and for 8 h on day 4 and day 6 postweaning. Mean serum LH concentrations increased (P<0.01) in weaned cows (W) from 0.55 +/- 0.01 ng/ml at time of calf removal to 1.3 +/- 0.04 ng/ml 48 h afterwards. Comparable LH concentrations for suckled cows (S) were 0.65 +/- 0.08 ng/ml and 0.62 +/- 0.03 ng/ml respectively. Average serum LH concentrations at 48 h after weaning were greater (P<0.01) for W cows than S cows and a treatment by time interaction occurred (P<0.01) with serum LH concentrations increasing (P<0.01) from time of calf removal to 48 h after calf removal in W cows. Frequency of LH peaks increased (P<0.01) in W cows and by 48 h after weaning was greater (P<0.01) in W cows than in S cows. Magnitude of LH peaks did not differ between the two groups. Serum cortisol concentrations were not different between W and S cows except for a transient elevation (P<0.01) in W cows from 7.6 +/- 0.9 ng/ml to 11.9 +/- 1.0 ng/ml 9 to 12 h after calf removal. Since serum LH concentrations were increased in W cows but not in S cows at 48 h and serum cortisol concentrations increased transiently in W cows we suggest that circulating cortisol levels may not be a physiological inhibitor of LH secretion in the suckled postpartum beef cow.     

Effects of transcutaneous thermal and electrical stimulation of the teat on pituitary luteinizing hormone, prolactin and oxytocin secretion in ovariectomized, estradiol-treated beef cows following acute weaning

The objective of the study was to determine if chronic electrical or thermal stimulation of sensory neurons on the surface of the teat is able to activate pathways that suppress the weaning-induced increase in luteinizing hormone (LH) secretion in beef cows. Treatment groups (n = 5 per group) consisted of: 1) control suckled (CS); 2) weaned plus electrical stimulation of the teat (ESTT); 3) weaned plus electrical stimulation of the tail (ESTL); 4) weaned plus thermal stimulation of the teat (TTT); 5) weaned plus thermal stimulation of the tail (TTL) and 6) weaned (WN). Cows were ovariectomized on Day 5 post partum (PP) and were treated with estradiol-17beta to maintain a constant tonic baseline. Beginning on Days 17 to 21 post partum, cows were suckled by their own calf (control), weaned or weaned and electrically or thermally stimulated for 10 minutes every 6 hours for 4 days. Chronic transcutaneous electrical and thermal stimulation of the teat or tail failed to impede the unambiguous rise (P < 0.001) in LH pulse frequency and amplitude following weaning. Positive and negative feedback of estradiol on LH secretion was not affected by treatments. Relatively consistent episodes of oxytocin and prolactin release were observed following control-suckling, but responses to electrical and thermal stimulation were inconsistent. Chronic electrical or thermal stimulation of teat-specific or nonspecific loci did not attenuate heightened secretion of LH after weaning. The results are further evidence against a role for mammary somatosensory neurons in the suckling-mediated inhibition of LH secretion.     

Effects of suckling and of a long interval after ovariectomy on hypothalamo-hypophyseal responsiveness to naloxone, morphine and GnRH in beef cows

The response of serum luteinizing hormone (LH) to morphine, naloxone and gonadotropin-releasing hormone (GnRH) in ovariectomized, suckled (n=4) and nonsuckled (n=3) cows was investigated. Six months after ovariectomy and calf removal, the cows were challenged with 1mg, i.v. naloxone/kg body weight and 1 mg i.v. morphine/kg body weight in a crossover design; blood was collected at 15-minute intervals for 7 hours over a 3-day period. To evaluate LH secretion and pituitary responsiveness, 5 mug of GnRH were administered at Hour 6 on Day 1. On Days 2 and 3, naloxone or morphine was administered at Hour 3, followed by GnRH (5 mug/animal) at Hour 6. Mean preinjection LH concentrations (3.6 +/- 0.2 and 4.7 +/- 0.2 ng/ml), LH pulse frequency (0.6 +/- 0.1 and 0.8 +/- 0.1 pulses/hour) and LH pulse amplitude (2.9 +/- 0.5 and 2.9 +/- 0.6 ng/ml) were similar for suckled and nonsuckled cows, respectively. Morphine decreased (P < 0.01) mean serum LH concentrations (pretreatment 4.2 +/- 0.2 vs post-treatment 2.2 +/- 0.2 ng/ml) in both suckled and nonsuckled cows; however, mean serum LH concentrations remained unchanged after naloxone. Nonsuckled cows had a greater (P < 0.001) LH response to GnRH than did suckled cows (area of response curve: 1004 +/- 92 vs 434 +/- 75 arbitrary units). We suggest that opioid receptors are functionally linked to the GnRH secretory system in suckled and nonsuckled cows that had been ovariectomized for a long period of time. However, gonadotropin secretion appears not to be regulated by opioid mechanisms, and suckling inhibits pituitary responsiveness to GnRH in this model.     

Naloxone evokes large-amplitude GnRH pulses in luteal-phase ewes

Ewes were sampled during the mid-late luteal phase of the oestrous cycle. Hypophysial portal and jugular venous blood samples were collected at 5-10 min intervals for a minimum of 3 h, before i.v. infusions of saline (12 ml/h; N = 6) or naloxone (40 mg/h; N = 6) for 2 h. During the 2-h saline infusion 2/6 sheep exhibited a GnRH/LH pulse; 3/6 saline infused ewes did not show a pulse during the 6-8-h portal blood sampling period. In contrast, large amplitude GnRH/LH pulses were observed during naloxone treatment in 5/6 ewes. The mean (+/- s.e.m.) amplitude of the LH secretory episodes during the naloxone infusion (1.07 +/- 0.11 ng/ml) was significantly (P less than 0.05) greater than that before the infusion in the same sheep (0.54 +/- 0.15 ng/ml). Naloxone significantly (P less than 0.005) increased the mean GnRH pulse amplitude in the 5/6 responding ewes from a pre-infusion value of 0.99 +/- 0.22 pg/min to 4.39 +/- 1.10 pg/min during infusion. This episodic GnRH secretory rate during naloxone treatment was also significantly (P less than 0.05) greater than in the saline-infused sheep (1.53 +/- 0.28 pg/min). Plasma FSH and prolactin concentrations did not change in response to the opiate antagonist. Perturbation of the endogenous opioid peptide system in the ewe by naloxone therefore increases the secretion of hypothalamic GnRH into the hypophysial portal vasculature. The response is characterized by a large-amplitude GnRH pulse which, in turn, causes a large-amplitude pulse of LH to be released by the pituitary gland.     

Mechanical masking of neurosensory pathways at the calf-teat interface: endocrine, reproductive and lactational features of the suckled anestrous cow

A mammary somatosensory mask was employed in suckled anestrous beef cows to attenuate signals that were hypothesized to play a direct regulatory role in postpartum anestrus. Cows (n = 20) were randomly assigned to one of three treatment groups on Days 15 to 20 postcalving. The three treatments were: 1) masked (n = 7); 2) suckled (negative control, n = 6); and 3) weaned (positive control, n = 7). Four layers of surgical glove latex were used to cover the teats and ventro-lateral prominence of the udder of masked cows with a nonhardening, nontoxic adhesive (Day 0). Masks were designed to prevent direct contact between the skin of the teat/udder and the mouth of the calf and to allow normal suckling and milk removal. Masks were left in place for 7 d, with calves in the weaned group removed to a remote location for 7 d. Calves in the suckled group were allowed ad libitum suckling. Calves in the masked group tended (P < 0.1) to suckle longer than calves in the suckled control group (11.3 +/- 1.3 vs. 7.8 +/- 1.3 min/suckle) posttreatment; however, suckling frequency and calf weight gains did not differ due to treatment. Weaned cows exhibited a four-fold increase (P < 0.01) in the frequency of luteinizing hormone (LH) pulses on Day 2 relative to suckled and masked cows. The percentage of animals ovulating within 12 d after treatment differed (P < 0.05) and was 100, 50 and 0% for weaned, suckled and masked cows, respectively. Presence of the latex mask allowed essentially normal suckling and lactation, but failed to attenuate (and may have potentiated) the negative effects of suckling on secretory patterns of LH, ovulation and estrus.     

Effect of naloxone and beta-casomorphin on the hypothalamic-pituitary-luteinizing hormone axis in vitro.

The effect of naloxone and beta-casomorphin on luteinizing hormone (LH) release from pituitary cell aggregates, obtained by three-dimensional culture, with or without mediobasal hypothalamic fragments was studied in vitro. Short-term naloxone perifusion at a concentration of 10(-5)M did not modify either basal or LHRH-stimulated LH release from the pituitary cell aggregates. In contrast, a 12-min naloxone perifusion at the same concentration caused an increase in LH release in the mediobasal hypothalamic-pituitary cell aggregate axis. This increase was rapid (12-16 min after time pulse), marked [up to 10 times (p less than 0.004) the initial base line], short (return to the base line secretion 32-40 min after the beginning of the time pulse) and dose-dependent, with a rise greater than 1000% at a concentration of 10(-4) (p less than 0.006). The same effect was observed when a second pulse was applied 48 min after the first one. LH release induced by naloxone was antagonized 56 +/- 2% (p less than 0.03) by beta-casomorphin (an exogenous opiate) at a concentration of 10(-5) M. beta-casomorphin alone did not modify LH basal secretion, but inhibited 25.1 +/- 2.4% (p less than 0.008) LH release enhanced by LHRH. These results indicate that naloxone, an opiate antagonist, markedly increases LH release via a mu-type opioid receptor mechanism at the hypothalamic level only, during short-term exposure.     

Effect of suckling on basal and GnRH-induced LH release in post-partum dairy buffaloes

Suckling, a common practice in smallholder dairy-farming systems in the developing world, delays the onset of post-partum ovarian activity in dairy buffalo. The present study was designed to assess the effect of suckling on pituitary function in lactating buffaloes 25-35 days post-partum. Six suckled and nine non-suckled buffaloes were challenged intravenously with a bolus injection of GnRH (20microg buserelin acetate; Receptal). Heparinized venous blood samples were collected at 15min intervals for 2h before and up to 4h after GnRH for luteinizing hormone (LH) estimation. Pretreatment basal LH concentrations were similar in the suckled (0.6+/-0.2ng/ml) and the non-suckled (0.5+/-0.1ng/ml) buffaloes. All but one suckled buffaloes released a LH surge, starting 15-60min post-GnRH treatment, which lasted for 180-225min. While one suckled buffalo did not respond to GnRH, the LH response in the remaining suckled buffaloes was significantly less than in the non-suckled buffaloes in terms of peak LH concentrations (14.3+/-2.7ng/ml versus 26.2+/-4.3ng/ml) and area under the LH curve (1575.6+/-197.4mm(2) versus 2108.9+/-323.9mm(2)). The LH response was least in suckled buffaloes challenged with GnRH while in the luteal phase of an oestrus cycle and with plasma progesterone concentration >1ng/ml. In conclusion, suckling suppressed pituitary responsiveness to exogenous GnRH challenge in post-partum buffaloes.     

Pituitary and ovarian function in postpartum beef cows. I. Effect of suckling on serum and follicular fluid hormones and follicular gonadotropin receptors

The effect of suckling on serum and follicular fluid hormones and on follicular gonadotropin receptors was studied. Sixteen anestrous postpartum cows were assigned to 1 of 2 groups: suckled (S) or weaned (W). All calves were allowed to suckle ad libitum from parturition to 21 days postpartum when calves from W cows were weaned. All cows were ovariectomized on Day 25 postpartum. W cows had more (P less than 0.01) pulses of LH during the 96-h period from weaning until ovariectomy than S cows (6.3 vs. 1.3 pulses). Serum concentrations of prolactin (Prl), estrone (E1), estradiol-17 beta (E2) and progesterone (P) were not different (P greater than 0.10) between groups. Furthermore, there were n differences (P greater than 0.10) in follicular in contents of luteinizing hormone (LH), E1, E2 and P between the treatment groups. However, follicular fluid content of Prl was greater (P less than 0.05) in the W cows than in the S cows (123 vs. 65.1 ng/cow). The number of follicular LH receptors was greater (P less than 0.05) in the W cows than in the S cows (71.1 vs. 48.3 fmoles/mg protein) although the number of follicular follicle-stimulating hormone (FSH) receptors was not different (P greater than 0.10) between W cows and S cows (1531 vs. 1862 fmoles/mg protein). There were no correlation between serum hormone concentrations and follicular fluid hormone content; however, the numbers of follicular LH receptors and follicular fluid Prl content were highly correlated in the W cows (r = 0.85; P less than 0.05). It is concluded that removal of the suckling stimulus increases pulsatile LH release and the accumulation of Prl in the follicular fluid. These factors, either together or separately, may at least in part be responsible for the increase in follicular LH receptor concentrations that were observed in the W cows.     

Independence of progesterone blockade of the luteinizing hormone surge in ewes from opioid activity at naloxone-sensitive receptors.

Fifteen ovariectomized ewes were treated with implants (s.c.) creating circulating luteal progesterone concentrations of 1.6 +/- 0.1 ng ml-1 serum. Ten days later, progesterone implants were removed from five ewes which were then infused with saline for 64 h (0.154 mol NaCl l-1, 20 ml h-1, i.v.). Ewes with progesterone implants remaining were infused with saline (n = 5) or naloxone (0.5 mg kg-1 h-1, n = 5) in saline for 64 h. At 36 h of infusion, all ewes were injected with oestradiol (20 micrograms in 1 ml groundnut oil, i.m.). During the first 36 h of infusion, serum luteinizing hormone (LH) concentrations were similar in ewes infused with saline after progesterone withdrawal and ewes infused with naloxone, but with progesterone implants remaining (1.23 +/- 0.11 and 1.28 +/- 0.23 ng ml-1 serum, respectively, mean +/- SEM, P greater than 0.05). These values exceeded circulating LH concentrations during the first 36 h of saline infusion of ewes with progesterone implants remaining (0.59 +/- 0.09 ng ml-1 serum, P less than 0.05). The data suggested that progesterone suppression of tonic LH secretion, before oestradiol injection, was completely antagonized by naloxone. After oestradiol injection, circulating LH concentrations decreased for about 10 h in ewes of all groups. A surge in circulating LH concentrations peaked 24 h after oestradiol injection in ewes infused with saline after progesterone withdrawal (8.16 +/- 3.18 ng LH ml-1 serum).(ABSTRACT TRUNCATED AT 250 WORDS)     

Ovarian steroid involvement in endogenous opioid modulation of LH secretion in seasonally anoestrous mature ewes

In June, 16 mature ewes were ovariectomized and allocated to four groups: 1, saline; 2, naloxone; 3, progesterone implant plus naloxone; 4, oestrogen implant plus naloxone. Steroids were implanted at the time of ovariectomy. At 5 days after ovariectomy, the animals were intravenously infused with saline for 8 h and naloxone (50 mg/h) in saline for 8 h the following day. Three intact ewes were given naloxone in a similar way. During infusions and for 8 h on the day after naloxone, jugular venous blood samples were taken every 15 min and assayed for LH. Naloxone resulted in significant increases in mean LH concentration (P less than 0.01), LH episode frequency and episode height (P less than 0.05) in Group 3 ewes, but was without effect in any other group. These results provide evidence that the progesterone status of the ewe affects its response to naloxone, that progesterone negative feedback on LH release may be mediated by an opioid system, and that increased oestradiol negative feedback during seasonal anoestrus is unlikely to work via increased opioid inhibition of LH.     

Opioid inhibition of luteinizing hormone secretion in the postpartum lactating sow

Twelve lactating sows were used at 22.4 +/- 0.8 days postpartum to determine whether endogenous opioid peptides (EOP) are involved in the suckling-induced inhibition of luteinizing hormone (LH) secretion. Four sows each received either 1, 2, or 4 mg/kg body weight of naloxone (NAL), an opiate antagonist, in saline i.v. Blood was collected at 15-min intervals for 2 h before and 4 h after NAL treatment. All sows were then given 100 micrograms gonadotropin-releasing hormone (GnRH) in saline i.v., and blood samples were collected for an additional h. Pigs were weaned after blood sampling. At 40 h after weaning, sows were treated and blood samples collected as during suckling. Serum concentrations of LH after treatment with NAL were similar for all doses; therefore, the data were pooled across doses. During suckling, serum concentrations of LH were 0.41 +/- 0.04 ng/ml before NAL treatment, increased to 0.65 +/- 0.08 ng/ml at 30 min after NAL treatment, and remained elevated above pretreatment concentrations for 120 min (p less than 0.05). Naloxone failed to alter serum concentrations of LH after weaning. These data indicate that EOP may be involved in the suckling-induced suppression of LH secretion and that weaning may either decrease opioid inhibition of LH secretion or decrease pituitary LH responsiveness to endogenous GnRH released by NAL.     

Diurnal variation in the response of anoestrous ewes to the ram effect

The re-introduction of rams after a period of separation was used to stimulate LH secretion and induce ovulation in seasonally anovulatory ewes maintained under natural photoperiod. In 2 experiments, the rams were introduced in the morning or the evening to test for diurnal variations in responsiveness to the treatment. In the first experiment, with Romanov ewes, the ram-induced increase in tonic LH secretion was significantly earlier in the ewes treated (N = 6) at 07:30 h (mean +/- s.e.m. delay to first pulse: 20 +/- 6 min) than in those (N = 5) treated at 19:30 h (66 +/- 15 min; P = 0.006). The pulse interval after the ram effect was significantly shorter in ewes that subsequently ovulated (120 +/- 10 min) than in ewes that did not ovulate (288 +/- 108 min; P = 0.043). There was a significant decline in pulse amplitude from 6.7 +/- 1.2 to 3.4 +/- 0.6 ng/ml (both groups combined) after the introduction of rams (P = 0.040). Of the 11 ewes, 7 subsequently ovulated and a preovulatory LH surge was observed in 6 of these 30-36 h after ram introduction. In the second experiment, with seasonally anoestrous Préalpes-du-Sud ewes, the effect of the timing of the introduction of rams on the periovulatory events was tested. The delay to the onsets of oestrus and the LH surge was not affected, but the ovulation rate was higher after ram introduction in the morning (1.42) than in the evening (1.14). In the 12-h period before the introduction of the rams in the first experiment, there was a difference between the groups in the secretion of LH, but the existence of diurnal rhythms in the concentrations of LH or FSH were not confirmed in a later study in which 7 ewes were sampled every 20 min for 36 h. In contrast, there was a distinct diurnal variation in the secretion of prolactin, with the highest values being recorded at night and the lowest around midday (P = 0.025). The rise and fall in prolactin values did not appear to coincide with dawn or dusk.(ABSTRACT TRUNCATED AT 400 WORDS)     

Use of naloxone challenge to predict sexual performance of rams before the fall breeding season

Serving capacity tests (SCT) identify rams as sexually active, sexually inactive, or low sexual performers. Labor and time requirements make SCT impractical and expensive. Therefore, a hormone-based test for libido was developed using the responses of LH and testosterone (T) after naloxone. This test effectively identified sexually active and inactive rams during the breeding season (November and December). The current study evaluated the effectiveness of this method at a different time of the year. Objectives were to determine if the blood test can detect differences in libido during late July and August, and to characterize LH and T profiles up to 120 min postnaloxone treatments at this time of year. Two experiments were conducted using i.v. naloxone at 0.75 mg/kg BW. Sexually active (SA) and sexually inactive (SIA) rams were identified using SCT. In Experiment 1, 16 SA and 18 SIA rams of several breeds were identified based on SCT conducted before naloxone tests. In Experiment 2, 72 Suffolk x white face rams were given SCT after naloxone tests and were of unknown sexual activity at the time of the naloxone tests. Mixed model analyses for repeated measures with repeated factors (month and sample time) and a covariate for mean LH or T before naloxone treatment were used for LH and T data. Proc Logistics modeled probabilities that rams were sexually active. The LH response to naloxone increased but did not differ (P > 0.09) by ram class (SA or SIA) or month. The T response peaked 75-90 min after naloxone treatment and was still elevated at 120 min. Testosterone differed (P < 0.03) for ram class by month by time in Experiment I and differed (P < 0.01) between months in Experiment 2. Proc Logistics incorrectly indicated that SA and SIA rams were all sexually active. This study indicates that the naloxone challenge test cannot discriminate between SA and SIA rams during July and August. Further research is required to determine the time of year when this sire identification test for libido has significant accuracy.     

The effects of nutrition on the reproductive performance of Finn x Dorset ewes. II. Post-partum ovarian activity, conception and the plasma concentration of progesterone and LH.

After lambing forty-five ewes were allocated to three groups, two of sixteen and one of thirteen ewes. The lambs of the two groups of sixteen ewes were weaned on Day 1 after lambing and the ewes were fed a diet of 100% (Group H) or 50% (Group R) of maintenance energy requirements. The thirteen ewes in the third group (Group L) suckled twin lambs and were fed freely. During the first 3 weeks after lambing, oestrus was observed for 11/16 (Group H) and 8/16 (Group R) ewes; of the ewes which had shown oestrus in the two groups, ovulation occurred in 5/8 and 5/7 respectively. Only 1/13 Group-L ewes showed oestrus and ovulated during the same period. The mean plasma concentrations of progesterone and LH were unaffected by the treatments and were around 0-4 and 1-5 ng/ml, respectively. Restricted feeding had no effect on oestrus, ovulation or the hormone levels during the oestrus cycle following synchronization. The onset of oestrus and the start of the preovulatory discharge of LH were 3 and 6 hr later, respectively, in the lactating ewes (Group L) than in those in Groups H and R. Ewes in Group L also had a higher ovulation rate, 2-8 +/- 0-2 versus 2-1 +/- 0-2 (P less than 0-05). Restricted feeding reduced the number of ewes lambing; only 1/11 ewes in Group R, considered to have conceived because of the presence of high progesterone levels 17 days after mating, subsequently lambed compared with 6/12 in Group H and 5/9 in Group L.     

Fluctuation in responsiveness of LH and lack of responsiveness of FSH to prolonged infusion of morphine and naloxone in the ewe

In ewes in the mid-luteal phase, LH pulse frequency (P less than 0.01) and amplitude (P less than 0.05) increased during a 24 h infusion of naloxone (0.5 mg/kg/h) compared to a 24 h infusion of vehicle (mean +/- s.e.m.; 0.25 +/- 0.03 vs 0.14 +/- 0.01 pulses/h and 0.84 +/- 0.08 vs 0.55 +/- 0.08 ng/ml serum, respectively). The increase in pulse amplitude was immediate, but was less (P less than 0.05) during the second 12 h, compared to the first 12 h, of naloxone infusion (0.52 +/- 0.14 vs 0.98 +/- 0.08 ng/ml serum). Oestradiol concentrations were higher (P less than 0.01) during naloxone than during control infusion (5.63 +/- 0.26 vs 4.13 +/- 0.15 pg/ml serum). In ovariectomized ewes in the breeding season, LH pulse frequency was lower (P less than 0.01) during a 24 h infusion of morphine (0.5 mg/kg/h) than during a 24 h infusion of vehicle (mean +/- s.e.m.; 1.17 +/- 0.08 vs 1.71 +/- 0.06 pulses/h). We conclude that long-term infusion of naloxone results in a sustained increase in LH pulse frequency but only a transient elevation in pulse amplitude. No effects on FSH secretion were noted. LH secretion was sensitive to morphine in the absence of ovarian steroids, suggesting that ovarian steroids are not required for the presence of functional opioid receptors capable of modulating LH release.     

Changes in pulsatile LH secretion after ovariectomy in Ile-de-France ewes in two seasons

Two experiments were conducted in Ile-de-France ewes to study changes in pulsatile LH secretion in ewes ovariectomized during anoestrus or during the midluteal phase of the oestrous cycle. In Exp. 1, blood samples were taken every 20 min for 12 h the day before ovariectomy (Day 0). After ovariectomy, samples were taken every 10 min for 6 h (10 ewes per group), on Days 1, 3, 7 and 15. In Exp. 2 samples were taken every 10 min for 6 h (10 ewes per group) on Days 7, 15, 30, 60, 90, 120, 150 and 180 after ovariectomy. Further samples were taken (5 ewes per group) at 9 and 12 months after ovariectomy. There were significant interactions between season and day of sampling for the interval between LH pulses in both experiments. LH pulse frequency increased within 1 day of ovariectomy and the increase was more rapid during the breeding season. There were clear seasonal differences in pulse frequency in Exp. 2. Compared with ewes ovariectomized in anoestrus, pulse frequency was significantly higher for ewes ovariectomized in the breeding season, from Day 7 until Day 120. Once pulse frequency had increased in ewes about the time of the normal breeding season, pulse frequency remained high and subsequent seasonal changes were greatly reduced. Pulse amplitude increased immediately after ovariectomy to reach a maximum on Day 7 and there were no differences between season of ovariectomy in the initial changes in amplitude. In Exp. 2, changes in amplitude followed changes in pulse interval and there was a significant interaction between season and day of sampling. There were no significant effects of season on nadir LH concentrations which increased throughout the duration of the experiments. These results show that, in ovariectomized ewes, LH pulse frequency observed on a given day depends on time after ovariectomy, season at the time of sampling and on previous exposure of ewes to stimulatory effects of season. The direct effects of season on LH pulse frequency and seasonal changes in sensitivity to steroid feedback may contribute to control of the breeding season and their relative contributions to the beginning and end of the breeding season may differ.     

A priming effect of naloxone on in vitro LHRH release from the hypothalamus of mid-luteal ewes

The possible involvement of endogenous opioid peptides (EOPs) in LHRH release from hypothalami of ewes during the breeding season was investigated using an in vitro perifusion system. Hypothalami were procured in December from ovariectomized (OVX; 62-65 days before the experiment; n = 6) and mid-luteal (ML; n = 7) Western White-Face ewes. Hypothalami were mid-sagitally sectioned into halves containing the preoptic area, mediobasal hypothalamus, and infundibulum (median eminence). The left half (treated) received two 30-min challenges (beginning at 130 and 250 min, respectively, after onset of perifusion) of 500 microM naloxone (NAL) followed by a 30-min 60-mM potassium (K) challenge (at 370 min after onset of perifusion). The right half served as the control, receiving only K at the same time as the treated tissue. Both NAL challenges elicited (p less than 0.05) LHRH release from tissues of both ML and OVX ewes. Release of LHRH by hypothalami from ML, but not from OVX, ewes was greater (p less than 0.01) after the second than after the first NAL challenge. These results are consistent with the view that an inhibitory opioid influence exists on LHRH release from ovine hypothalami. The release of LHRH in response to NAL was dependent on the ovarian status in vivo since the priming effect of NAL on subsequent NAL-induced LHRH release occurred only from the hypothalami of ML ewes. We suggest from these results that EOPs may modulate LHRH release from ovine hypothalami in an ovarian steroid-dependent and independent manner.     

Evidence that the corpus luteum of pregnancy contributes to the control of tonic secretion of LH in the ewe

Concentrations of LH and FSH were measured in blood samples collected from the jugular vein at 20-min intervals for 7 h (09:00-16:00 h) on Days 60, 80, 100 and 120 of pregnancy in 5 intact ewes and 5 from which the CL had been excised on Day 70. In the 5 intact ewes, plasma LH concentrations remained low and unchanged between Days 60 and 120. During this period, pulsatile release of LH occurred irregularly and infrequently. Removal of the CL resulted in an increase in the basal values of LH and in the frequency and amplitude of LH pulses. Concentrations of FSH were relatively constant in all stages of pregnancy examined and were similar in both groups of ewes. These results show that (1) LH concentrations are low during the second half of pregnancy; and (2) LH, but not FSH, increases after CL excision, presumably by removing some luteal factor inhibitor of LH secretion.     

Effects of naloxone or transient weaning on secretion of LH and prolactin in lactating sows

Sows (N = 16) were infused intravenously for 8 h with saline or naloxone (200 mg/h) or their litters were transiently weaned for 8 h. Before infusion, 200 mg naloxone were administered to elevate quickly concentrations of naloxone. Blood samples were collected from sows at 15 min intervals for 24 h, beginning 8 h before and continuing until 8 h after imposition of treatments during the middle 8-h segment. Frequency of episodic release of LH and concentrations of prolactin were similar before, during and after infusion of saline. Average concentration of LH was greater during the last than during the middle 8-h segment when sows were given saline. Frequency of episodic release of LH increased and concentrations of prolactin decreased during infusion of naloxone or transient weaning; however, average concentration of LH increased during transient weaning, but not during infusion of naloxone. After transient weaning or infusion of naloxone, frequency of release of LH decreased, returning to pretreatment values in sows infused with naloxone but remaining above pretreatment values in sows subjected to transient weaning. At the resumption of suckling by litters in sows subjected to transient weaning, prolactin increased to levels not different from those observed during the 8-h pretreatment segment. Prolactin did not increase until 4-5 h after cessation of naloxone infusion. We conclude that continuous infusion of naloxone altered secretory patterns of LH and prolactin. Collectively these results provide evidence that the immediate effects of weaning on LH and prolactin in sows are mediated in part through a mechanism involving endogenous opioid peptides.     

Circulating concentrations of luteinizing hormone, testosterone, and growth hormone after naloxone treatment in sexually mature boars

The effects of naloxone, an antagonist of opioid peptides, on circulating concentrations of luteinizing hormone (LH), testosterone, and growth hormone (GH) were determined in sexually mature boars. Blood samples were collected at 15-min intervals for three hr from five crossbred boars. Two hr after initiation of blood sampling, boars received an i.v. challenge of naloxone (1 mg/kg body weight; n=2) or 0.9% saline (n=3). Twenty-four hr later the experiment was repeated, but boars that previously received naloxone received saline and vice versa. A time by treatment interaction (p=0.09) was detected for concentrations of LH in serum, and levels of LH were greater (p<0.03) after treatment with naloxone compared to saline. Concentrations of testosterone in serum were affected by time (p<0.01), but not treatment (p= 0.59) or treatment by time (p=0.74). A treatment by time interaction (p=0.02) was detected for serum GH concentrations. Levels of GH increased in saline-treated boars (p<0.01), but not in boars receiving naloxone (p>0.1). Our results are consistent with the theory that opioid peptides suppress LH secretion and stimulate GH release in sexually mature boars.