The stimulation of testosterone and LH secretion by synthetic GnRH in the male Cape porcupine

The effects of GnRH stimulation on plasma testosterone and luteinizing hormone (LH) levels in Cape porcupine males were examined by analysing plasma collected before and after an intravenous injection of GnRH. In six mature males and one subadult, which were given an intravenous injection of 0,5 ml saline, levels of plasma testosterone and LH did not increase. Four weeks later an intravenous GnRH challenge (40 μ?) caused plasma testosterone to rise three-fold and LH to rise 10-15-fold within 180 min in five of the mature males. Peaks of plasma testosterone and LH occurred 90 and 120 min, respectively, after stimulation, and baseline and peak levels of both hormones were significantly related.     

Effect of photoperiod and castration on prolactin, testosterone and luteinizing hormone concentrations in male calves

After 8 wk exposure to 8 h of light per day, prolactin (PRL) averaged 18.3 ng/ml of serum in eight male calves. Four calves then received 16 h of light per day; 6 wk later (age 14 wk) PRL averaged 93.8 ng/ml of serum, whereas PRL averaged 36.9 ng/ml of serum in four calves maintained under 8 h of daily light. By wk 20, PRL was not different in calves exposed to 16 or 8 h of daily light, averaging 34.7 and 17.2 ng/ml serum. Testosterone averaged .43 ng/ml of serum at wk 8 but was greater at wk 14 in calves receiving 16 h of light daily when compared with controls receiving 8 h of light (1.92 vs. .97 ng/ml of serum). Testosterone concentrations were not different between photoperiod treatments at wk 20. Luteinizing hormone (LH) concentrations were unaffected by photoperiod. In a second experiment, four male calves were castrated at approximately 2 wk of age while four similar controls were left gonadally intact. After 8 wk exposure to 8 h of light per day, PRL averaged 12.3 ng/ml of serum in all calves. After 6 wk exposure to 16 h of light per day, PRL in serum increased in castrates to 48.0 ng/ml and in controls to 59.8 ng/ml. We conclude that serum concentrations of PRL and testosterone, but not LH, increased in bull calves receiving 16 h of light daily relative to calves receiving 8 h of light, and that the PRL response to photoperiod is independent of the testes. However, 16 h light-induced stimulation of serum concentrations of prolactin is not maintained indefinitely.     

Effect of GnRH and hCG administration on plasma LH and testosterone concentrations in normal stallions, aged stallions and stallions with lack of libido

Gonadotrophin-releasing hormone (GnRH) (a single intravenous injection with 0.042 mg busereline acetate) was administered to control stallions (n=5), aged stallions (n=5) and stallions with lack of libido (n=5). Jugular blood samples were taken at -10, 0, 10, 20, 40 and 80 minutes after treatment and measured for luteinizing hormone (LH) and testosterone concentrations. A single intravenous injection of hCG (3000 IE) was given 1 day later. Venous blood samples were taken at -60, 0, 15, 30, 60, 120, and 240 minutes after treatment and measured for the testosterone concentration. The experiment was performed in the breeding season. There was a wide variation between stallions in basal concentrations of LH and testosterone. The treatment groups all showed a significant increase in LH and testosterone concentrations after treatment with GnRH. There was a significant difference (P<0.05) between the control, the lack of libido stallions and the aged stallions in the production of LH before and after stimulation with GnRH. The aged stallions had higher basal LH concentrations. GnRH induced a rise in plasma LH in all groups, but the greatest response was observed in aged stallions. No response to GnRH was seen with respect to plasma testosterone. There was an increase in plasma testosterone following hCG; however, this increase was very small in aged stallions. After stimulation with hCG the control and lack of libido stallions had a significant increase (P<0.05) in testosterone production. In conclusion, stimulation with either GnRH or hCG can be a valuable method to test whether the function of the stallion's reproductive endocrine system is optimal.     

Effects of GnRH treatment on initiation of pulses of LH,LH release,and subsequent concentrations of progesterone

Progesterone is essential for establishment and maintenance of pregnancy. One proposed method to increase progesterone is administering GnRH at insemination. However, this method has resulted in conflicting results. Therefore, 2 experiments were conducted to evaluate how administering GnRH at insemination affected pulses of luteinizing hormone (LH) and subsequent progesterone. In Experiment 1, cows were allotted to 2 treatments: (1) GnRH (100 μg) given approximately 12 h after initiation of estrus (n = 5); and (2) Control (n = 5). Blood samples were collected at 15-min intervals for 6 h at 12 (blood sampling period 1), 26 (blood sampling period 2), 40 (blood sampling period 3), 54 (blood sampling period 4), and 68 (blood sampling period 5) h after onset of estrus. Daily blood samples were collected for 17 d. In Experiment 2, cows were allotted into 2 treatments: GnRH administered 10 to 11 h (n = 10) or 14 to 15 h (n = 10) after onset of estrus. Daily blood samples were collected for 17 d. Cows treated with GnRH tended (P ≤ 0.075) to have greater LH release during blood sampling period 1, tended (P = 0.095) to have fewer pulses during blood sampling period 2, tended (P = 0.067) to have greater concentrations of progesterone, and had an earlier (P = 0.05) increase in progesterone than control cows. Cows treated with GnRH 10 to 11 h after onset of estrus had greater (P = 0.01) progesterone and an earlier (P = 0.04) increase in progesterone than cows treated 14 to 15 h. In conclusion, timing of GnRH treatment following onset of estrus influenced pulses of LH and subsequent progesterone.     

A single administration of the GnRH antagonist acyline inhibits basal and GnRH-stimulated serum testosterone concentrations in male dogs

The objective of this study was to describe testosterone (T) response to GnRH challenge in antagonist-treated dogs over a 30-day period. Eight mongrel dogs were randomly assigned to either the GnRH antagonist acyline 330 μg/kg sc (ACY; n = 4) or a placebo group (PLA; n = 4). The dogs were serially challenged with the GnRH agonist, buserelin 0.2 μg/kg sc on days -1, 1, 3, 7, 10, 14, 21 and 30. On these days, blood samples for T determinations were collected before (-30 min) and 60, 120 and 180 min after the agonist injection. Basal (-30 min) and post-GnRH agonist stimulation T values were compared by anova for repeated measures. Before treatments (day -1), there were no differences in basal T serum concentrations between groups (p > 0.1). After treatments, basal T showed a significant interaction between treatment and day (p < 0.05). Furthermore, when both groups were analysed independently, basal T varied in the ACY (p < 0.01) but not in the PLA group (p > 0.1). On day -1, before treatments, the stimulation tests had only a time effect (p = 0.05) although on days 1 (p < 0.01), 3 (p < 0.01), 7 (p < 0.01), 10 (p < 0.01) and 14 (p < 0.05), the response to the agonist differed between groups, becoming similar on days 21 (p > 0.05) and 30 (p > 0.05). It was concluded that, in dogs, a single administration of the GnRH antagonist prevented canine gonadal axis to physiologically respond to agonistic challenge during 14 days.     

Pituitary responsiveness to GnRH, hypothalamic content of GnRH and pituitary LH and FSH concentrations immediately preceding puberty in gilts

To determine whether pituitary concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH) or hypothalamic content of gonadotropin releasing hormone (GnRH) change before puberty, 40 prepubertal gilts averaging 7 mo of age were slaughtered before or on the second, third or fourth day after relocation and boar exposure. Some gilts responded to relocation and boar exposure as indicated by swollen vulvae, turgid uteri and enlarged ovarian follicles at the time of slaughter. Pituitary concentrations of LH and FSH and hypothalamic content of GnRH were similar between gilts that responded to relocation and boar exposure and gilts that did not respond. In addition, boar exposure and relocation had no effect on pituitary concentrations of LH and FSH or on hypothalamic content of GnRH. To determine whether pituitary responsiveness to GnRH changes before puberty, a third experiment was conducted in which 72 gilts were injected with 400 micrograms of GnRH either before or on the second, third or fourth day after relocation and boar exposure. In gilts that subsequently responded (i.e., ovulated) as a result of relocation and boar exposure, pituitary responsiveness to GnRH was reduced as compared with gilts that failed to ovulate after relocation and boar exposure. Peak concentrations of serum LH after GnRH injection were 4.6 +/- 1.3 vs 9.8 +/- .8 ng/ml for responders vs nonresponders. Peak serum FSH after GnRH injection was also lower for responders than for nonresponders (29.5 +/- 4.2 vs 41.2 +/- 2.4 ng/ml). When compared with controls, relocation and boar exposure did not significantly affect GnRH-induced release of LH and FSH.(ABSTRACT TRUNCATED AT 250 WORDS)     

Energy balance, first ovulation and the effects of naloxone on LH secretion in early postpartum dairy cows

Relationship among energy balance (EB), first ovulation, circulating metabolites and opioid involvement in peripheral LH concentrations were assessed in 40 multiparous Holstein cows assigned randomly to an experiment with a 2 x 2 factorial arrangement of treatments. Cows were either milked normally (2x/d) and expected to be in negative EB (LAC) or were milked for only 1 d postpartum (PP) to ensure a rapid return to positive EB (NONLAC). Blood samples were collected from NONLAC cows every 12 min for 16 h on d 7, 10, and 14 PP and from LAC cows on d 7, 14, 21 and 28 PP. Within each group, half the cows received naloxone (50 mg/h) and half received saline during h 9 to 16 of each frequent sampling series. The mean number of LH pulses/8 h for NONLAC cows was 4.3, 4.7 and 5.3 at 7, 10 and 14 d PP and for LAC cows 4.3, 5.7, 6.4 and 7.0 at 7, 14, 21 and 28 d PP, respectively. The LH pulse frequency was not different between NONLAC and LAC cows at 7 and 14 d PP, yet NONLAC cows had fewer days to first ovulation (P less than .01). Naloxone did not affect any parameters of LH secretion in either group. Following parturition, NONLAC cows averaged 4.0 d to negative EB nadir and 14.3 d to first ovulation. The LAC cows averaged 13.6 d to negative EB nadir and 27.0 d to first ovulation. Days PP to first ovulation were highly correlated (r = .85) with days PP to negative EB nadir.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of epinephrine, norepinephrine and(or) GnRH on serum LH in prepuberal beef heifers

Forty prepuberal Simmental X Brahman-Hereford heifers were utilized to determine the effects of epinephrine (E), norepinephrine (NE), gonadotropin releasing hormone (GnRH) or combinations of GnRH + E and GnRH + NE on serum luteinizing hormone (LH) concentrations. Animals were assigned randomly to one of five treatments with four replicates/treatment. Treatments consisted of I) 100 micrograms GnRH at time 0 (n = 8); II) 50 mg NE at time -15 and 0 (n = 8); III) 50 mg E at time -15 and 0 (n = 8); IV) 100 micrograms GnRH at time 0, plus 50 mg NE at time -15 and 0 (n = 8) and V) 100 micrograms GnRH at time 0, plus 50 mg E at time -15 and 0 (n = 8). All treatment compounds were administered im in 2 ml physiological saline and blood samples were collected via tail vessel puncture at -30, -15, 0, 15, 30, 45, 60, 90, 120, 180, 240, 300 and 360 min from GnRH injection. Treatment with NE or E alone had no effect (P greater than .10) on serum LH during the sampling period. The initial LH release to GnRH was altered (P less than .05) by concomitant treatment with NE (treatment IV) or E (treatment V). Magnitude of the LH release was reduced (P less than .01) by treatment V. Area under the LH surge was reduced (P less than .05) by treatment IV (NE) and V (E).     

Relationships of gonadotropins, testosterone, and cortisol in response to GnRH and GnRH antagonist in boars selected for high and low follicle-stimulating hormone levels

Considerable variation exists in the serum levels of gonadotropins in boars; this results in differential testicular function. Boars (Chinese Meishan, European White composite, and crosses of the two breeds) selected for high and low circulating FSH concentrations were used to define possible differences in pituitary sensitivity to GnRH and GnRH antagonist and gonadal and adrenal responses. After a 2-h pretreatment sampling period, boars were injected with GnRH or GnRH antagonist and repetitively sampled via jugular cannula for changes in serum concentrations of FSH, LH, testosterone, and cortisol. In response to varying doses of GnRH or GnRH antagonist, FSH, LH, or testosterone changes were not different in high- or low-FSH boars. Declines in LH after GnRH stimulation were consistently faster in boars selected for high FSH. Chinese Meishan boars had considerably higher cortisol concentrations than White composite boars (132.2 +/- 28.5 vs 67.4 +/- 26.8 ng/mL, respectively; P < .01). When select high- and low-gonadotropin Meishan:White composite crossbreds were sampled, cortisol levels were elevated but comparable between the two groups (126.5 +/- 13.7 vs 131.4 +/- 13.4 ng/mL, respectively). After GnRH antagonist lowered LH concentrations, administration of hCG resulted in increased testosterone and cortisol concentrations. Although testosterone concentrations remained high for 30 h, cortisol concentrations returned to normal levels within 10 h after hCG injection. The mechanism by which boars selected for high gonadotropins achieve increased levels of LH and FSH may not be due to differences in pituitary sensitivity to GnRH but to differences in clearance from the circulation.     

Temporal effects of estradiol (E) on luteinizing hormone-releasing hormone (LHRH) and LH release in castrated male sheep

We tested the hypothesis that rapidly expressed inhibitory effects of estradiol (E) on luteinizing hormone (LH) release in the male are attributable, in part, to suppression of luteinizing hormone-releasing hormone (LHRH) release. Hypophyseal-portal cannulated, castrated male sheep were infused with E (15 ng/kg/hr) or vehicle. Portal and jugular blood samples were collected at 10-min intervals for 4 hr before, and for either 12 hr (E, n = 4; vehicle, n = 4) or 24 hr (E, n = 8; vehicle, n = 3) after the start of infusion. In animals sampled for 16 hr, temporal changes in both LHRH and LH were assessed. In animals sampled for 28 hr, only LH data were analyzed. Before either the 12-hr or 24-hr infusion, LHRH and/or LH mean concentrations, pulse amplitude and interpulse interval (IPI) did not differ between E- and vehicle-infused animals. In animals sampled for 16 hr, no effects of time or steroid × time interactions were detected for mean LHRH and LHRH pulse amplitude; however, both were greater (P < 0.01) in vehicle-infused than in E-infused males. LHRH IPI was unaffected by infusion. In contrast, both mean LH and LH pulse amplitude declined (P < 0.01) within 4–8 hr after the start of E infusion, whereas mean LH IPI was unaffected. In animals sampled for 28 hr, an effect of time (P < 0.01) and a steroid × time interaction (P < 0.01) was detected for mean LH, and there was an effect of time (P < 0.01) on LH pulse amplitude. Mean LH IPI was not affected. Our results show that in male sheep E rapidly reduces LH release in the absence of a detectable change in LHRH release.     

Relationships among LH, FSH and prolactin secretion, storage and response to secretagogue and hypothalamic GnRH content in ovariectomized pony mares administered testosterone, dihydrotestosterone, estradiol, progesterone, dexamethasone or follicular fluid

Thirty-five ovariectomized pony mares were used to study the relationships among luteinizing hormone (LH), follicle stimulating hormone (FSH) and prolactin (PRL) concentrations in blood (secretion), in pituitary (storage) and in blood after secretagogue administration, as well as the content of gonadotropin releasing hormone (GnRH) in hypothalamic areas, under various conditions of steroidal and nonsteroidal treatment. Five mares each were treated daily for 21 d with vegetable shortening (controls), testosterone (T; 150 micrograms/kg of body weight, BW), dihydrotestosterone (DHT; 150 micrograms/kg BW), estradiol (E2; 35 micrograms/kg BW), progesterone (P4; 500 micrograms/kg BW), dexamethasone (DEX; 125 micrograms/kg BW) or charcoal-stripped equine follicular fluid (FF; 10 ml). Secretagogue injections (GnRH and thyrotropin releasing hormone, TRH, at 1 and 4 micrograms/kg of BW, respectively) were given one d prior to treatment and again after 15 d of treatment. Relative to controls, treatment with T, DHT and DEX reduced (P less than .05) LH secretion, storage and response to exogenous GnRH, whereas treatment with E2 increased (P less than .05) these same characteristics. Treatment with P4 reduced (P less than .05) only LH secretion. Treatment with T, DHT, E2 and DEX reduced (P less than .05) FSH secretion, whereas treatment with P4 increased (P less than .05) it and FF had no effect (P greater than .1). All treatments increased (P less than .05) FSH storage, whereas only treatment with T and DHT increased (P less than .05) the FSH response to exogenous GnRH. Other than a brief increase (P less than .05) in PRL secretion in mares treated with E2, secretion of PRL did not differ (P greater than .1) among groups. Only treatment with E2 increased (P less than .01) PRL storage, yet treatment with T or DHT (but not E2) increased (P less than .05) the PRL response to exogenous TRH. Content of GnRH in the body and pre-optic area of the hypothalamus was not affected (P greater than .1) by treatment, whereas treatment with T, E2 and DEX increased (P less than .1) GnRH content in the median eminence. For LH, secretion, storage and response to exogenous GnRH were all highly correlated (r greater than or equal to .77; P less than .01). For FSH, only storage and response to exogenous GnRH were related (r = .62; P less than .01). PRL characteristics were not significantly related to one another. Moreover, the amount of GnRH in the median eminence was not related (P greater than .1) to any LH or FSH characteristic.     

Effects of plasma progesterone concentrations on LH release and ovulation in beef cattle given GnRH

The effects of plasma progesterone concentrations on LH release and ovulation in beef cattle given 100 microg of GnRH im were determined in three experiments. In Experiment 1, heifers were given GnRH 3, 6 or 9 days after ovulation; 8/9, 5/9 and 2/9 ovulated (P<0.02). Mean plasma concentrations of progesterone were lowest (P<0.01) and of LH were highest (P<0.03) in heifers treated 3 days after ovulation. In Experiment 2, heifers received no treatment (Control) or one or two previously used CIDR inserts (Low-P4 and High-P4 groups, respectively) on Day 4 (estrus=Day 0). On Day 5, the Low-P4 group received prostaglandin F(2alpha) (PGF) twice, 12 h apart and on Day 6, all heifers received GnRH. Compared to heifers in the Control and Low-P4 groups, heifers in the High-P4 group had higher (P<0.01) plasma progesterone concentrations on Day 6 (3.0+/-0.3, 3.0+/-0.3 and 5.7+/-0.4 ng/ml, respectively; mean+/-S.E.M.) and a lower (P<0.01) incidence of GnRH-induced ovulation (10/10, 9/10 and 3/10). In Experiment 3, 4-6 days after ovulation, 20 beef heifers and 20 suckled beef cows were given a once-used CIDR, the two largest follicles were ablated, and the cattle were allocated to receive either PGF (repeated 12h later) or no additional treatment (Low-P4 and High-P4, respectively). All cattle received GnRH 6-8 days after follicular ablation. There was no difference between heifers and cows for ovulatory response (77.7 and 78.9%, P<0.9) or the GnRH-induced LH surge (P<0.3). However, the Low-P4 group had a higher (P<0.01) ovulatory response (94.7% versus 61.1%) and a greater LH surge of longer duration (P<0.001). In conclusion, although high plasma progesterone concentrations reduced both GnRH-induced increases in plasma LH concentrations and ovulatory responses in beef cattle, the hypothesis that heifers were more sensitive than cows to the suppressive effects of progesterone was not supported.     

Effects of progesterone and weaning on LH and FSH responses to naloxone in postpartum beef cows

Two experiments were conducted to evaluate the effects of naloxone, an endogenous opioid receptor antagonist, on LH and FSH secretion in postpartum beef cows. In Experiment 1, 24 cows were divided into three equal groups. On day 15 postpartum, all cows were bled for 8 hr at 10 min intervals to evaluate LH secretory parameters. On day 18 postpartum, three treatments were administered: (a) saline at 0730 and 1130 hr; (b) 275 mg naloxone at 0730 and 1130 hr; (c) naloxone as in (b) above, plus this group was also treated with 50 mg progesterone (P4) twice daily from day 16 to day 19. In each treatment, jugular vein samples were collected at 10 min intervals from 0800 to 1600 hr. On day 19 the same treatments were administered at the same times, however, all cows were given 25 micrograms GnRH at 1200 hr to evaluate the LH secretory response. Naloxone increased mean LH concentration (P less than .05) and tended to increase pulse amplitude and frequency compared to controls. However, the most dramatic difference was due to P4 treatment which suppressed mean LH, pulse amplitude and frequency. Treatments had no effect on LH secretion in response to a 25 micrograms dose of GnRH. In Experiment 2, the effects of suckling on the naloxone response were examined in 16 postpartum cows. On day 21 postpartum, blood was collected at 10 min intervals for 8 hr and then calves were removed from half the cows. After 3 days of calf removal, all cows were sampled at 10 min intervals for 4 hr; then naloxone was injected after each 10 min sample at a dose rate of 200 mg/hr (33 mg per injection). Naloxone treatment and sampling continued for an additional 8 hr. Calf removal alone had very little effect on LH pulsatility. However, naloxone resulted in increased pulse frequency and mean LH compared to the control period. We conclude that LH release in the early postpartum cow is partially regulated by endogenous opioid peptides. We were unable to detect any effects on FSH secretion nor on pituitary sensitivity to exogenous GnRH.