Simultaneous measurement of luteinizing hormone-releasing hormone and luteinizing hormone during estradiol-induced luteinizing hormone surges in the ovariectomized ewe

Sequential bleeding and push-pull perfusion of the hypothalamus were used to characterize luteinizing hormone (LH) and LH-releasing hormone (LHRH) release in ovariectomized (OVX) ewes after injection of corn oil or estradiol benzoate (EB). Push-pull cannulae were surgically implanted into the stalk median eminences of 24 OVX ewes. Seven to 14 days later each of 20 animals was given an i.m. injection of 50 micrograms EB. Blood samples and push-pull perfusate were collected at 10-min intervals for 6-12 h beginning 12-15 h after EB injection. Four OVX ewes were given i.m. injections of corn oil 7 days after implantation of push-pull cannulae. Blood samples and push-pull perfusate were collected at 10-min intervals for 4 h between 18 and 22 h after injection of corn oil. Luteinizing hormone remained below 2 ng/ml throughout most of the sampling periods in 9 of 20 EB-treated ewes. In 5 of these 9 LHRH also was undetectable, whereas in 4 LHRH was detectable (1.84 +/- 0.29 pg/10 min), but did not increase with time. Preovulatory-like surges of LH occurred in 11 EB-treated ewes, but LHRH was undetectable in 5. In 4 of 6 ewes showing LH surges and detectable LHRH, sampling occurred during the onset of the LH surge.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of active immunization of ram lambs and bull calves against synthetic luteinizing hormone releasing hormone

Ram lambs and bull calves were immunized against LH-RH by injections given in weeks 0, 6, 12 and 28 (ram lambs, week 0 = 16 to 20 weeks of age) and weeks 0, 6, 12 and 18 (bull calves, week 0 = approximately 4 weeks of age). The testis size of LH-RH-immunized animals was significantly less than that of controls from week 13 onwards in ram lambs and from week 15 onwards in bull calves. When ram lambs were sampled in week 17 and bull calves in week 20, mean plasma gonadotrophin and testosterone concentrations were consistently lower in LH-RH-immunized animals than in controls. Single intravenous injection of synthetic LH-RH or an analogue of LH-RH in week 27 failed to induce LH or testosterone responses in LH-RH-immunized ram lambs. Motile semen samples could not be obtained from any of the LH-RH-immunized ram lambs in weeks 24, 25 and 26 or from 7 of 10 in week 72, but samples of moderate motility were obtained in week 72 from three rams in which LH-RH antibody titres had fallen. No attempt was made to obtain semen from bull calves. After castration there was no increase in plasma LH in LH-RH-immunized rams and only a small increase in LH-RH-immunized bull calves. Mean testis weight was significantly lower in LH-RH-immunized animals than in controls of both species. Thus the normal development of the reproductive system in ram lambs and bull calves was blocked by active immunization against LH-RH. Some evidence was obtained for natural reversal of the effects with time and falling antibody titres. These findings demonstrate the potential of LH-RH immunization as an alternative to castration.     

Maturational changes in opioidergic control of luteinizing hormone and follicle-stimulating hormone in ram lambs

Stimulation by naloxone, an opioid antagonist, of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) secretion was examined in spring-born crossbred ram lambs raised under natural photoperiod. Vehicle (n = 6) or 1 mg naloxone/kg vehicle (n = 6) was injected (i.m.) 3 times at 2-h intervals at 5, 10 and 15 weeks of age and 4 times at 2-h intervals at 20, 25, 30 and 35 weeks of age. Blood samples were taken every 12 min for 6 h at 5, 10 and 15 weeks of age and for 8 h at 20, 25, 30 and 35 weeks of age. Naloxone had no effect on age at sexual maturity (controls 239 +/- 23 days; naloxone 232 +/- 33 days). The only significant (P less than 0.05) effect of naloxone on FSH was a greater pulse amplitude in 10-week-old treated lambs than in control lambs. Naloxone treatment resulted in greater LH pulse amplitude at 5 and 10 weeks of age (P less than 0.05), lower basal serum concentration of LH at 10 weeks of age (P less than 0.05), greater LH pulse frequency at 25 weeks of age (P less than 0.05), and greater mean serum concentrations of LH, basal LH and LH pulse amplitude at 35 weeks of age (P less than 0.01) than in the controls. In both groups of lambs, mean and basal FSH, and LH and FSH pulse amplitude were highest at 5 weeks of age and fell with age. LH pulse amplitude was lowest at 35 weeks of age (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of active immunization of the ram and bull against luteinizing hormone

Active immunization of ram lambs and bull calves against highly purified LH results in the production of antibodies which bind radiolabeled ovine LH. Antibody titers were not dependent upon the dose (0.1 or 1.0 mg) of antigen given but were found to be greater when the antigen consisted of LH conjugated to a carrier protein; in this case human serum globulin (hSG). The presence of LH antibodies in ram sera seemed of little consequence. The presence of antibodies in sera of immunized bulls, however, appeared to neutralize endogenous LH sufficiently to suppress serum testosterone and reduce testicular size. Androgen deficiency, produced by immunization against oLH-hSG, resulted in atrophy of the testes, seminal vesicles and epithelial cells lining both epididymides and seminal vesicles. Reduced weight gain and lack of development of the secondary sexual characteristics were at levels expected of surgical castrates (steers). In summary, active immunization of young beef bulls against LH and in particular, oLH-hSG, results in a castration-like response which has been compared with that achieved in other species by luteinizing hormone releasing hormone (LHRH) immunocastration. The success of LH immunocastration in field trials with young bulls and investigations aimed toward reversibility are yet to be determined.     

Ontogeny of growth hormone, prolactin, luteinizing hormone, and testosterone secretory patterns in the ram

The ontogenetic changes that occur in secretory patterns of growth hormone (GH), prolactin (Prl), luteinizing hormone (LH), and testosterone (T) in rams maintained in constant photoperiod were examined. Nine ram lambs were moved to individual pens in a controlled environment (12L: 12D cycle; 18-24 degrees C temperature) at 66 days of age. Blood samples were collected via indwelling cannulae at 15-min intervals for an 8-h period at 80, 136, 192, 248, and 304 days of age. Plasma concentrations of GH, Prl, LH, and T were quantitated and parameters of the secretory patterns determined. Mean concentration of GH tended to decline with age, probably because the amplitude of secretory peaks was significantly reduced with age. There were no age-associated changes in basal concentration of GH or incidence of GH peaks. There was an increase in Prl secretion (as estimated by mean concentration) at 136 and after 248 days of age. Significant age-associated changes occurred in all parameters of LH and T secretion. At the younger ages, testosterone concentrations were low and LH concentrations were elevated. At the older ages the relationship was reversed, with LH low and testosterone high. There were no significant correlations between frequency and magnitude of LH and T peaks. The significant correlations present among parameters of LH and T secretion were between basal concentration of LH and overall mean concentration and basal concentration of T. These results suggest that LH may not be the sole tropic stimulator of acute T secretion.     

Hypothalamic sites of catecholamine inhibition of luteinizing hormone in the anestrous ewe.

Norepinephrine (NE) and dopamine (DA) actively inhibit the release of LH in anestrous ewes. This can be detected as an increase in LH pulse frequency following i.v. injection of NE and DA antagonists. The objective of this study was to determine the sites of these inhibitory actions in the ovine hypothalamus by using local administrations of the NE antagonist, phenoxybenzamine (PBZ), or the DA antagonist, pimozide (PIM), into specific hypothalamic areas. Each neurotransmitter antagonist was administered via a chronically implanted steel guide tube into either the preoptic area (POA), retrochiasmatic area (RCh), or the median eminence region (ME). Blood samples were taken every 15 min for 2 h before and 4 h during implantation of these drugs and were analyzed for LH and prolactin by RIA. Control (no treatment) samples were obtained similarly from the same animals on another day. Placement of PBZ into the POA significantly increased LH pulse frequency and mean LH concentrations over control values whereas PIM did not. In contrast, PIM significantly increased LH pulse frequency and mean LH concentrations when placed in the ME or in the RCh, but PBZ did not. No effects of PIM on prolactin concentrations were detected. These results suggest that an NE neural system operates in the POA and that a DA system acts in the medial basal hypothalamus (RCh or ME) to suppress GnRH pulse frequency in the ovary-intact anestrous ewe.     

Effects of pentobarbital anesthesia on tonic luteinizing hormone secretion in the ewe: evidence for active inhibition of luteinizing hormone in anestrus

In the ewe, seasonal anestrus appears to result from two effects of inhibitory photoperiod: 1) estradiol gains the capacity to suppress luteinizing hormone (LH) pulse frequency and hence becomes a potent inhibitor of tonic LH secretion and 2) a steroid-independent decrease in LH pulse frequency occurs in ovariectomized ewes. In this study, we have obtained evidence, using pentobarbital anesthesia, that both these actions of photoperiod reflect the activation, in anestrus, of an inhibitory neural system. Administration of pentobarbital to intact anestrous ewes produced a dramatic, 3-fold increase in LH pulse frequency during the 6 h of anesthesia. In contrast, during the breeding season, pentobarbital inhibited LH pulse frequency in luteal phase animals. There was also a seasonal variation in the effects of pentobarbital in ovariectomized ewes. During the breeding season this drug again suppressed LH secretion, inhibiting both LH pulse amplitude and frequency. In anestrus, pentobarbital also suppressed pulse amplitude, but it produced a transitory increase (lasting 3 h) in pulse frequency. To account for the stimulatory actions of pentobarbital, we propose that in anestrus, but not the breeding season, LH pulse frequency is held in check by a set of estradiol-sensitive inhibitory neurons. Further, we suggest that these neurons are activated by inhibitory photoperiod and account for both the steroid-dependent and steroid-independent actions of photoperiod.     

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.     

Release of luteinizing hormone in prepubertal and middle-aged ovariectomized rats

Concentrations of circulating LH were determined in conscious, free-moving ovariectomized rats. All of the animals had been ovariectomized at 24 days of age. Between 30 and 90 days there was an increase in mean blood LH concentrations; a more vigorous pulsatile release of LH characterized by an increase in amplitude and frequency of LH release; and an elevated responsiveness to LHRH administration. Rats which had been ovariectomized for 1 year still had elevated blood LH levels but had episodic pulses of reduced amplitude and a decrease in responsiveness to LHRH. These data suggest that important alterations occur with age in the neuroendocrine mechanisms responsible for the release of LH.     

The mechanism of the stimulation of LH production by synthetic LH-releasing hormone (LH-RH) in tissue culture

Synthetic LH-RH was found to stimulate production of LH by human female adenohypophysis in monolayer culture. This effect occurs at 0.30 μg/2 ml LH-RH. New messenger-RNA synthesis does not have to occur to stimulate the production of LH by the action of synthetic LH-RH in cultures of under 4 days. In cultures of over 4 days, this synthesis must occur in order for LH to be produced by the action of LH-RH. However, new DNA synthesis does not have to occur to stimulate the production of LH by the action of synthetic LH-RH.