In utero programming of sexually differentiated gonadotrophin releasing hormone (GnRH) secretion

It has long been recognised that steroids can have both organisational and activational effects on the reproductive neuroendocrine axis of many species, including the sheep. Specifically, if the ovine foetus is exposed to testosterone during a relatively short 'window' of in utero development (from approximately day 30-90 of a 147 day pregnancy) the neural mechanisms regulating gonadotrophin releasing hormone (GnRH) secretion become organised in a male-specific manner. In post-natal life the consequences of foetal androgen exposure are sexually differentiated responses of the GnRH neuronal network to activation by factors such as photoperiod and ovarian steroid hormones. Studies in the gonadectomized lamb have demonstrated that elevated concentrations of oestrogen (E) are unable to trigger a preovulatory-like GnRH surge in the male and the androgenized ewe lamb. Further, these animals have markedly reduced sensitivity to the inhibitory actions of progesterone on tonic GnRH release compared with normal ewes. The reasons for these abnormal steroid feedback mechanisms may reside in sexually dimorphic inputs to the GnRH neurone, including those from oestrogen-receptive neurones in the arcuate nucleus that synthetize the neuropeptide, neurokinin B (NKB). The consequences of in utero androgen exposure are reflected in a progressive and dramatic impairment of fertility in the ovary-intact ewe.     

Induction of ovulation with gonadotrophin releasing hormone and progesterone in seasonally anestrous mares

Five seasonally anestrous mares were treated with a regimen of gonadotrophin releasing hormone and progesterone in an attempt to induce estrus and ovulation. The treatment induced follicular activity and estrus in all mares. Two of the five mares ovulated but none conceived.     

Efficacy of human chorionic gonadotrophin and gonadotrophin releasing hormone for hastening ovulation in thoroughbred mares

Plasma progesterone levels were measured daily to determine the accuracy of diagnosing ovulation by rectal palpation carried out every other day; 81.5 per cent mares injected with human chorionic gonadotrophin showed increases of progesterone more than 1 ng/ml by 72 h after injection compared with 65 per cent of mares injected with gonadotrophin releasing hormone (GnRH) or saline. Mating at ovulation achieved a 74 per cent pregnancy rate in mares given hCG compared with 50 per cent given GnRH and 45 per cent controls. Diagnosis of ovulation per rectum on the basis of a pit in the ovarian surface or the presence of a soft friable structure was found to be accurate in 91 per cent of cases. Diagnosis based on the presence of a firm plum-like structure or the disappearance of a previously identified follicle at a given site was 60 per cent accurate. In cases of rectal diagnosis of non-ovulation 38 per cent had actually ovulated based on a rise in progesterone more than 1 ng/ml and a further 19 per cent showed a rise in progesterone by 24 h after the diagnosis. The use of hCG may improve conception rates by synchronisation of ovulation and mating and where rectal palpation of the ovaries remains the only basis for timed mating.     

Luteinizing hormone (LH) response to different doses of synthetic gonadotropin releasing hormone (GnRH) in prepubertal gilts

The object of this investigation was to study luteinizing hormone (LH) response to different doses of synthetic gonadotropin-releasing hormone (GnRH) in prepubertal gilts. Four crossbred prepubertal gilts, 128–134 days old and body weight 57–63 kg, were used in this study. Four doses, 0. 5, 25 and 125 μg, of GnRH were administered via a jugular vein catheter in a latin square design. Each treatment consisted of 3 injections at 90 min intervals. Frequent blood samples were taken during a period of 90 min before up to 90 min after treatment. Total LH responses were measured from post-treatment samples as the area under the curve above base level obtained from pre-treatment samples. A positive relationship between GnRH dose and LH release was obtained in all gilts, except for 1 treatment given to a gilt with high plasma level of oestradiol-17β on the day of treatment. This study has demonstrated the responsiveness of the pituitary gland by LH release to different doses of GnRH in 4.5-month-old prepubertal gilts.     

青年期及产蛋期绍兴鸭和康贝尔鸭血清促性腺激素释放激素浓度的比较

采用放射免疫分析法 (RIA)测定了不同日龄 (60 ,10 0 ,2 0 0和 380日龄 )绍兴鸭和卡基 康贝尔鸭 (KhakiCampbellDuck)血清中促性腺激素释放激素 (GnRH)浓度 ,试图通过比较不同品种、性别和日龄鸭之间GnRH浓度的变化 ,探讨GnRH与产蛋性能的关系。结果表明 :绍兴鸭和康贝尔鸭母鸭血清中GnRH浓度呈现类似的年龄性变化 ,60日龄时GnRH浓度较低 ,10 0日龄时最高 ,2 0 0日龄时显著下降 ,380日龄时回升至较高水平。公鸭血清中GnRH浓度的年龄性变化与母鸭相似 ,10 0日龄时最高 ,2 0 0日龄时最低     

Suppression of testicular function using two dose rates of a reversible water soluble gonadotrophin releasing hormone (GnRH) vaccine in colts

Objective: To investigate the effect of two dose rates (200 and 400 ng) of a gonadotrophin releasing hormone (GnRH) vaccine on testicular function. Design: A vaccination dose rate experiment. Procedure: Two injections were administered 4 weeks apart to six colts in each treatment group. To maintain immunosuppression until the end of the breeding season, a third injection was given if antibody titres fell below 1000. Results: Effective antibody titres were present for 12 to 27 weeks. Testosterone concentrations decreased from 2.22 to 0.31 nmol/L 6 weeks after primary vaccination. Androstenedione concentrations decreased from 1.78 to 0.28 nmol/L 5 weeks after vaccination. Testosterone and androstenedione concentrations above 0.69 and 0.87 nmol/L were attained 31 to 43 weeks after vaccination. Mean scrotal widths and lengths decreased over 29 weeks from 9.2 cm and 9.7 cm to 6.7 cm and 7.6 cm. At surgical castration these dimensions were 10.1 cm and 11.0 cm. Mean semen characteristics before vaccination and after recovery were: gel-free volume 16.5 and 13.5 mL, sperm concentration 295.5 times 10⁶ 315.6 times 10⁶/mL, total sperm per ejaculate 4041 times 10⁶ 4657 times 10⁶ live normal spermatozoa 32% and 60%. Histologically, the testes showed active spermatogenesis. The mean testicular parenchyma weights for the 200 and 400 mg groups were 129.0 g and 109.8 g. Daily sperm production per testis and per gram of testis for the 200 and 400 mg groups were 3.7 times 10⁸ 2.8 times 10⁶ 2.3 times 10⁸ 2.0 times 10⁶. Conclusions: Both dose rates suppressed testicular function. Data showed that the vaccine effects were reversible. Individual immune response was less varied in the 200 mg group. Further work is necessary to achieve a less variable response in the immunosuppression of testicular function.     

Gonadotropin releasing hormone (GnRH) induced luteinizing hormone (LH) secretion from perifused equine pituitaries.

In vitro responsiveness of the horse anterior pituitary (AP) gonadotropes to single and multiple GnRH challenges was examined. The pituitaries were collected from reproductively sound mares in estrus (n = 5) and diestrus (n = 5). Uniform 0.5 mm AP slices were subdivided using a 3 mm biopsy punch and then bisected for use in the perifusion chamber. Four bisected sections per chamber were perifused at 0.5 ml/min at 37 C for 560 min in Medium 199 saturated with 95% 0(2)/5% CO2. Ten minute fractions were collected after an initial 2 hr equilibration period. Four different treatment regimes of GnRH (10(-10) M) were evaluated: (A) three consecutive 10 min GnRH pulses separated by 80 and 100 min, respectively; (B) a single 120 min GnRH infusion; (C) a 10 min GnRH pulse followed 80 min later by a 120 min GnRH infusion and (D) two 10 min GnRH pulses separated by 60 min followed 80 min later by a 120 min GnRH infusion. Estimated total pituitary LH content was higher in estrous than diestrus mares (p less than 0.05). The total amount of LH released in response to GnRH tended to be greater in estrus than diestrus (p less than 0.1), whereas the percentage of LH released in estrus and diestrus was similar. An increase in the area under the LH response curve was noted with each successive 10 min pulse of GnRH during both estrus and diestrus (p less than 0.05), demonstrating a self-priming effect of GnRH. In addition, a significant increase in the peak LH amplitude (p less than 0.05) and the slope to peak amplitude (p less than 0.05) were observed for the 120 min GnRH pulse in regime C and D indicating that prior exposure to short-term pulses of GnRH increased the acute LH secretory response. These results suggest that in the cycling mare (1) the responsiveness of the pituitary (amount of LH released as percent of total LH) is similar in both estrus and diestrus, however, the magnitude of the LH response (total microgram amount of LH released) differs with the stage of the estrous cycle, being highest in estrus, and appears to be related, in part, to pituitary LH content and (2) GnRH self-priming occurs independently of the stage of the estrous cycle. Furthermore, we have demonstrated that the pulsatile mode of GnRH can act directly on the anterior pituitary to dictate the pulsatile release pattern of LH in the cycling mare.     

Response of the lactating and postweaning sow to gonadotropin releasing hormone (GnRH)

Clinical and endocrinological responses to administration of gonadotropin releasing hormone analog (LH-RH-A) during the lactation period and postweaning in the sow were investigated. Plasma LH concentrations in lactating sows rose immediately after administration of LH-RH-A. However, in postweaning sows the increase of LH level was more slowly. Three of 5 postweaning sows came into estrus and ovulated after LH-RH-A treatment. One sow exhibited a distinct LH response, but her ovaries remained quiescent. The remaining one with feeble estrus for a short period became cystic ovaries. Thus, LH response to GnRH in the sow seems to be higher during early lactation than at 2 days postweaning.     

A priming effect of gonadotrophin releasing hormone on luteinizing hormone secretion in the boar.

The possibility that gonadotrophin releasing hormone (GnRH) can prime the anterior pituitary to a second dose of GnRH resulting in a greatly enhanced secretion of luteinizing hormone was examined in three adult boars. Four experiments were conducted: saline injection followed one hour later by a second saline injection (control); 1 microgram of synthetic GnRH injection followed one hour later by saline injection; saline injection followed one hour later by GnRH injection; GnRH injection followed one hour later by a second GnRH injection. Immunoassayable levels of plasma luteinizing hormone resulting from GnRH plus GnRH treatment were significantly greater than the sum obtained when values from GnRH plus saline and saline plus GnRH were added. Testosterone values in plasma reached maximal concentrations about 60 minutes after peak values of luteinizing hormone were achieved. The results suggest that the first dose of GnRH, in addition to stimulating release of luteinizing hormone can also sensitize the gonadotrophs to a second dose of GnRH causing a significantly greater release of luteinizing hormone.     

Short-term changes in serum luteinizing hormone, ovarian response and reproductive performance following gonadotrophin releasing hormone treatment in postpartum dairy cows with retained placenta.

Thirty-six postpartum Holstein cows consisting of eighteen cows that shed the placenta soon after calving (NRP) and eighteen cows that retained placenta greater than 24 h (RP) were used. There were four treatment groups. Group I consisted of 9 NRP cows which received intramuscular injection of sterile saline on day 15 postpartum. The second group consisted of 9 NRPN cows which received 100 micrograms of gonadotrophin releasing hormone on day 15 postpartum (NRPT). The third group consisted of 9 RP cows which received saline on day 15 postpartum (RPN) and the fourth group consisted of 9 RP cows which received 100 micrograms of gonadotrophin releasing hormone on day 15 postpartum (RPT). Blood samples were collected once daily during the first month and once every other day during the second month postpartum. In addition fourteen cows (RPT, n = 7; NRPT, n = 7), were used to study short-term changes in serum luteinizing hormone concentrations following gonadotrophin releasing hormone treatment on day 15 postpartum. Blood samples were obtained from these cows every 15 min during 1 h before and 6 h after gonadotrophin releasing hormone administration. Sera from all samples were assayed for progesterone and luteinizing hormone concentrations. Starting from four days after calving rectal palpations and ultrasound examinations of the ovaries were carried out once every four days until day 28 postpartum in order to monitor ovarian changes. All cows were inseminated on the first estrus after day 60 postpartum and examined for pregnancy between 35 and 42 days after insemination.(ABSTRACT TRUNCATED AT 250 WORDS)     

Stimulation of annexin A5 expression by gonadotropin releasing hormone (GnRH) in the Leydig cells of rats

The distribution and regulation of annexin A5 expression, a gonadotropin releasing hormone (GnRH) receptor regulated protein in gonadotropes and luteal cells, in the testes of rats were examined. Immunocytochemical staining revealed high levels of annexin A5 in the Leydig and endothelial cells and lower levels in the primary spermatocytes and sperm. Hemicastration significantly increased the annexin A5 content of the remaining testis within 24 h. Annexin A5 immunoreactivity was increased mainly in interstitial tissues including the peritubular cells, while some spermatocytes also showed higher intensity of annexin A5 in the remaining testis. Administration of hCG (50 IU) enhanced the testicular content of annexin A5 after 24 h. This treatment expanded the area of interstitial tissue in the testis and increased annexin A5 immunoreactivity, but the area of the of endothelial cells was unchanged. Similarly, human chorionic gonadotropin (hCG) enhanced annexin A5 expression in a primary culture of testis cells that consisted of mainly interstitial cells. Because GnRH stimulates the expression of annexin A5 in the gonadotropes and luteal cells, we examined the effect of GnRH on annexin A5 expression in the testes. We found that des-Gly10 [Pro9]-GnRH ethylamide (100 nM), a GnRH agonist, increased annexin A5 expression in cultured testis cells and that Cetrorelix (100 nM), a GnRH antagonist, inhibited the effect of hCG on annexin A5 expression. These results suggest that pituitary luteinizing hormone promotes annexin A5 synthesis in Leydig cells and that this effect could be mediated by local GnRH in the testis.     

Clearance rate of gonadotrophin releasing hormone in peripheral plasma of the pig.

Gonadotrophin releasing hormone was administered as an intravenous bolus injection into four boars and four ovariectomized sows. Radioimmunoassay of concentrations of gonadotrophin releasing hormone in blood collected periodically after injection indicated a biexponential decline suggesting a rapid distribution to the extracellular fluid and a slower elimination by metabolism. A mean half-life value of 2.12 +/- 0.95 (SD) minutes was calculated for the first component and of 13.15 +/- 2.55 minutes was calculated for the second component of the decline in gonadotrophin releasing hormone concentrations. No significant difference was detected between boars and sows for half-life value of either component. In the four boars, luteinizing hormone values reached a peak in plasma 20 minutes after injection and that of testosterone at 90 minutes after gonadotrophin releasing hormone treatment.     

Reproductive performance in dairy cows following postpartum treatment with gonadotrophin releasing hormone and/or prostaglandin: a field trial.

Three hundred and five Holstein Friesian cows were given either 250 micrograms gonadotrophin releasing hormone (GnRH) or saline on day 15 postpartum followed by 500 micrograms cloprostenol or saline on day 24 postpartum. Four treatment groups were formed using random allocation: Group I -- placebo (Day 15)/placebo (Day 24), Group II -- GnRH (Day 15)/placebo (Day 24), Group III -- placebo (Day 15)/cloprostenol (Day 24), Group IV -- GnRH (Day 15)/cloprostenol (Day 24). Double blind techniques were used during the follow-up period. Rectal palpation, to assess uterine involution and ovarian activity was performed just prior to each treatment and again at 28 days postpartum. In addition blood samples were collected at 15, 24 and 28 days postpartum for measurement of plasma progesterone. There were no significant differences among treatment groups with respect to services per conception, number of heats detected before first service and culling for infertility. Cows treated only with GnRH had an increased calving to first estrus and calving to first breeding interval, and tended to have an increased calving to conception interval. Treatment with cloprostenol significantly decreased calving to conception and calving to first observed estrus intervals. Treatment with GnRH on day 15 postpartum resulted in a significant increase in the subsequent incidence of pyometra and prebreeding anestrus. On the other hand, cloprostenol treatment on day 24 postpartum resulted in a decreased incidence of pyometra, regardless of GnRH treatment and a decreased incidence of prebreeding anestrus in GnRH treated cows compared to cows receiving only GnRH at day 15 postpartum.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of growth hormone and gonadotrophin releasing hormone antagonists on ovarian follicle growth in sheep

Our objective was to determine the effects of the administration of growth hormone (GH) alone or plus teverelix, a gonadotrophin releasing hormone antagonist (GnRHa), on follicle development in sheep. Ewes were treated daily for 6 days by the intramuscular route with 15 mg of GH alone (GH group; n = 6) or combined with two subcutaneous doses of GnRHa (1.5 mg) on days 0 and 3 of GH treatment (GH/GnRHa group; n = 6); the control group (n = 6) received similar treatment with saline solution. Plasma follicle stimulating hormone levels were significantly lower in the GH/GnRHa group than in the control (P < 0.001) and GH groups (P < 0.05). The number of follicles > or =2 mm increased to reach significant differences with control (18.7 +/- 0.6) on day 4 in GH/GnRHa group (22.7 +/- 0.5, P < 0.001) and on day 5 in GH group (20.3 +/- 0.4 vs. 17.0 +/- 0.6, P < 0.05). These results indicate that GH and GnRHa may be useful for increasing the number of gonadotrophin-responsive follicles in the ovary. However, follicle function could be affected as both GH and GH/GnRHa groups showed lower plasma inhibin A concentrations than control sheep (90-110 pg/mL vs. 170-185 pg/mL, P < 0.005).