Negative feedback as an obligatory antecedent to the estradiol-induced luteinizing hormone surge in ovariectomized pigs

In ovariectomized pigs, estradiol treatment induces a preovulatory-like luteinizing hormone (LH) surge, but only after serum LH concentrations are suppressed for 48 h. This inhibition of LH release is attributable in large part to inhibition of gonadotropin-releasing hormone (GnRH) release. The present report examines the dependency of the estradiol-induced LH surge on this preceding phase of negative feedback. Ten ovariectomized gilts were given an i.m. injection of estradiol benzoate (10 micrograms/kg BW). Beginning at the time of estradiol treatment, 5 of these gilts received 1-microgram GnRH pulses i.v. every 45 min for 48 h, i.e. during the period of negative feedback. The remaining 5 control gilts received comparable infusions of vehicle. Estradiol induced the characteristic biphasic LH response in control gilts. On the other hand, the inhibitory LH response to estradiol was prevented and the ensuing LH surge was blocked in 4 of the 5 gilts given GnRH pulses during the negative feedback phase. These results indicate that suppressing release of GnRH and/or LH is an important antecedent to full expression of the LH surge in ovariectomized pigs. Assimilation of this observation with the existing literature provides novel insights into the neuroendocrine control of LH secretion in castrated and ovary-intact gilts.     

Luteinizing hormone secretion as affected by hypophyseal stalk transection and estradiol-17beta in ovariectomized gilts

The objectives were to determine hypothalamic regulation of pulsatile luteinizing hormone (LH) secretion in female pigs and the biphasic feedback actions of estradiol-17beta (E(2)-17beta). In the first study, the minimum effective dosage of E(2)-17beta that would induce estrus in ovariectomized gilts was determined to be 20microg/kg body weight. In the second study, ovariectomized gilts were assigned randomly on day 0 to treatments: (a) hypophyseal stalk transection (HST), (b) cranial sham-operated control (SOC), and (c) unoperated control (UOC). On day 3, gilts from each group received a single i.m. injection of either E(2)-17beta (20microg/kg body weight) or sesame oil. Blood was collected from an indwelling jugular cannula at 15min intervals for 3h before (day -2) and after treatment (day 2) from HST, SOC and UOC gilts. On day 3, blood was collected at 2h intervals for 12h after E(2)-17beta or sesame oil injection and at 4h intervals thereafter for 108h. Pulsatile LH secretion in all gilts 2 days after ovariectomy exhibited a frequency of 0.9+/-0.06peaks/h, amplitude of 1.3+/-0.13ng/ml, baseline of 0.8+/-0.07. Serum LH concentrations from SOC and UOC gilts were similar on day 2 and profiles did not differ from those on day -2. In HST gilts pulsatile LH release was abolished and mean LH concentration decreased compared with controls (0 versus 0.9+/-0. 06peaks/h and 0.77+/-0.03 versus 1.07+/-0.07ng/ml, respectively; P<0. 05). E(2)-17beta or sesame oil did not affect serum LH concentration in HST gilts, and LH remained constant throughout 120h (0.7+/-0. 07ng/ml). In SOC and UOC control gilts, E(2)-17beta induced a 60% decrease (P<0.05) in LH concentration within 12h, and LH remained low until 48h, then increased to peak values (P<0.05) by 72h, followed by a gradual decline to 120h. Although pituitary weight decreased 31% in HST gilts compared with controls (228 versus 332mg, P<0.05), an abundance of normal basophils was evident in coronal sections of the adenohypophysis of HST comparable to that seen in control gilts. The third and fourth studies determined that hourly i. v. infusions of LHRH (2microg) and a second injection of E(2)-17beta 48h after the first had no effect on the positive feedback action of estrogen in UOC. However, in HST gilts that received LHRH hourly, the first injection of E(2)-17beta decreased (P<0.05) plasma LH concentrations while the second injection of E(2)-17beta failed to induce a positive response to estrogen. These results indicate that both pulsatile LH secretion and the biphasic feedback action of E(2)-17beta on LH secretion depend on hypothalamic regulatory mechanisms in the gilts. The isolated pituitary of HST gilts is capable of autonomous secretion of LH; E(2)-17beta will elicit direct negative feedback action on the isolated pituitary gland if the gonadotropes are supported by exogenous LHRH, but E(2)-17beta at high concentrations will not induce positive feedback in isolated pituitaries. Thus, the direct effect of E(2)-17beta on the pituitary of monkeys cannot be mimicked in pigs.     

Failure of naloxone to stimulate luteinizing hormone secretion during pregnancy and steroid treatment of ovariectomized beef cows

The response of serum luteinizing hormone (LH) to naloxone, an opiate antagonist, and gonadotropin-releasing hormone (GnRH) was measured in cows in late pregnancy to assess opioid inhibition of LH. Blood samples were collected at 15-min intervals for 7 h. In a Latin Square arrangement, each cow (n = 6) received naloxone (0, 0.5, and 1.0 mg/kg BW, i.v.; 2 cows each) at Hour 2 on 3 consecutive days (9 +/- 2 days prepartum). GnRH (7 ng/kg body weight, i.v.) was administered at Hour 5 to all cows on each day. Mean serum LH concentrations (x +/- SE) before naloxone injection were similar (0.4 +/- 0.1 ng/ml), with no serum LH pulses observed during the experiment. Mean serum LH concentrations post-naloxone were similar (0.4 +/- 0.1 ng/ml) to concentrations pre-naloxone. Mean serum LH concentrations increased (p less than 0.05) following GnRH administration (7 ng/kg) and did not differ among cows receiving different dosages of naloxone (0 mg/kg, 1.44 +/- 0.20; 0.5 mg/kg, 1.0 +/- 0.1; 1.0 mg/kg, 0.9 +/- 0.1 ng/ml). In Experiment 2, LH response to naloxone and GnRH was measured in 12 ovariectomized cows on Day 19 of estrogen and progesterone treatment (5 micrograms/kg BW estrogen: 0.2 mg/kg BW progesterone) and on Days 7 and 14 after steroid treatment. On Day 19, naloxone failed to increase serum LH concentrations (Pre: 0.4 +/- 0.1; Post: 0.4 +/- 0.1 ng/ml) after 0, 0.5, or 1.0 mg/kg BW.(ABSTRACT TRUNCATED AT 250 WORDS)     

Anisomycin inhibition of estradiol-induced and progesterone-facilitated luteinizing hormone surges in the immature rat

These studies were designed to examine the effect of anisomycin, a potent and reversible inhibitor of protein synthesis with low systemic toxicity in rodents, on induction of luteinizing hormone (LH) surges by estradiol and their facilitation by progesterone. Immature female rats that received estradiol implants at 0900 h on Day 28 had LH surges approximately 32 h later (1700 h on Day 29). Insertion of progesterone capsules 24 h after estradiol led to premature (by 1400 h) and enhanced LH secretion. Protein synthesis was inhibited by 97%, 95%, 47%, and 16% in the hypothalamus-preoptic area (HPOA) and by 98%, 87%, 35%, and 0% in the pituitary at 30 min, 2 h, 4 h, and 6 h after s.c. injection of anisomycin (10 mg/kg BW), respectively. A single injection of anisomycin at 0, 3, 6, 9, 12, 24, 27, or 30 h after estradiol treatment significantly lowered serum LH levels at 32 h. The effect of injecting anisomycin at 0, 24, or 27 h was overridden by progesterone treatment at 24 h, but LH secretion was delayed serum LH levels were basal (10-30 ng/ml) at 1400 h but elevated (500-800 ng/ml) at 1700 h. Complete suppression of LH surges in estradiol-plus-progesterone-treated rats was achieved with 2 injections of anisomycin on Day 29 at 0900 h and again at 1200 h or 1400 h. Further experiments were designed to examine proteins that might be involved in anisomycin blockade of progesterone-facilitated LH surges. Intrapituitary LH concentrations at 1700 h on Day 29 were 70-80% higher (102 +/- 12.5 micrograms/pituitary) in rats that received 2 injections of anisomycin than in vehicle-treated controls (58.5 +/- 7.7 micrograms/pituitary). There were no significant effects of anisomycin on cytosol progestin receptors in the HPOA (7.1 +/- 1.5 fmol/tissue, anisomycin; 7.2 +/- 0.3, vehicle) or pituitary (8.3 +/- 1.3 fmol/tissue, anisomycin; 11.7 +/- 2.9, vehicle) at this time. The concentration of pituitary gonadotropin-releasing hormone receptors (GnRH-R), however, was significantly lower after anisomycin (265 +/- 30 vs. 365 +/- 37 fmol/mg protein) treatment. These results suggest that both estradiol-induced and progesterone-facilitated LH surges involve protein synthetic steps extending over many hours. Blockade of progesterone-facilitated LH surges by anisomycin appears to be due primarily to an effect on release of LH to which lowering of GnRH-R levels may contribute.     

Photoperiod and luteinizing hormone secretion in domestic and wild pigs

During seasonal anoestrus (long-days), oestradiol can effectively inhibit the pulsatile secretion of luteinizing hormone (LH) in sheep. The aim of our trial was to determine whether the same regulatory mechanism exists in the pig. Altogether, 20 ovariectomized and oestradiol-implanted gilts (16 domestic pigs, 4 European wild boars) were randomly allocated to two treatment groups. The first group was kept under a short-day light-dark cycle of 8L:16D, and the second group under a long-day light regime of 16L:8D. After a 6-week treatment period, blood samples were taken at 20-min intervals for 12h. After sampling, the light regimens were switched. Sampling was then repeated following another 6 weeks of treatment. In both treatment groups, 2.3 LH pulses occurred every 12h. The basal LH level was 0.7+/-0.4 ng/ml for the short-day group and 1.0+/-0.5 ng/ml for the long-day group. The mean LH level was 0.9+/-0.4 and 1.3+/-0.6 ng/ml and the LH pulse amplitude 0.5+/-0.4 and 0.6+/-0.5 ng/ml, respectively. The basal and mean LH levels were therefore lower in short-day gilts (P<0.05), while LH pulse amplitude and frequency remained unaffected by treatment. In conclusion, the 6-week period under two different light regimes resulted in higher basal LH concentration in long-day gilts but was not able to produce changes in LH frequency in prepubertal gilts.     

Different effects of subnormal levels of progesterone on the pulsatile and surge mode secretion of luteinizing hormone in ovariectomized goats

This study tested the hypothesis that endocrinological threshold levels of progesterone that induce negative feedback effects on the pulsatile and surge modes of LH secretion are different. Our approach was to examine the effects of subnormal progesterone concentrations on LH secretion. Long-term ovariectomized Shiba goats that had received implants of silastic capsules containing estradiol were divided into three groups. The high progesterone (high P) group received a subcutaneous implant of a silastic packet (50 x 70 mm) containing progesterone, and the low progesterone (low P) group received a similar implant of a small packet (25 x 40 mm) containing progesterone. The control (non-P) group received no treatment with exogenous progesterone. Blood samples were collected daily throughout the experiment for the analysis of gonadal steroid hormone levels and at 10-min intervals for 8 h on Days 0, 3, and 7 (Day 0: just before progesterone treatment) for analysis of the pulsatile frequency of LH secretion. Then estradiol was infused into the jugular vein of all animals at a rate of 3 microg/h for 16 h on Day 8 to determine whether an LH surge was induced. Blood samples were collected every 2 h from 4 h before the start of the estradiol infusion until 48 h after the start of the infusion. In each group, the mean +/- SEM concentration after progesterone implant treatment was 3.3 +/- 0.1 ng/ml for the high P group, 1.1 +/- 0.1 ng/ml for the low P group, and <0.1 ng/ml for the non-P group, concentrations similar to the luteal levels, subluteal levels, and follicular phase levels of the normal estrous cycle, respectively. The estradiol concentration ranged from 4 to 8 pg/ml after estradiol capsule implants in all groups. The LH pulse frequency was significantly (P < 0.05) suppressed on Day 3 (6.2 +/- 0.5 pulses/8 h) and on Day 7 (2.6 +/- 0.9 pulses/8 h) relative to Day 0 (9.0 +/- 0.5 pulses/8 h) in the high P group. In both the low P and non-P groups, however, the changes of pulsatile frequency of LH were not significantly different, and high pulses (7-9 pulses/8 h) were maintained on each of the 3 days they were tested. An LH surge (peak concentration, 100.3 +/- 11.0 ng/ml) occurred in all goats in the non-P group, whereas there was no surge mode secretion of LH in either the high P or the low P group. The results of this study support our hypothesis that the threshold levels of progesterone that regulate negative feedback action on the LH pulse and the LH surge are different. Low levels of progesterone, around 1 ng/ml, completely suppressed the LH surge but did not affect the pulsatile frequency of LH secretion.     

Influence of stage of the estrous cycle on endogenous opioid modulation of luteinizing hormone, prolactin, and cortisol secretion in the gilt

Fourteen gilts that had displayed one or more estrous cycles of 18-22 days (onset of estrus = Day 0) and four ovariectomized (OVX) gilts were treated with naloxone (NAL), an opiate antagonist, at 1 mg/kg body weight in saline i.v. Intact gilts were treated during either the luteal phase (L, Day 10-11; n = 7), early follicular phase (EF, Day 15-17; n = 3), or late follicular phase (LF, Day 18-19; n = 4) of the estrous cycle. Blood was collected at 15-min intervals for 2 h before and 4 h after NAL treatment. Serum luteinizing hormone (LH) concentrations for L gilts averaged 0.65 +/- 0.04 ng/ml during the pretreatment period and increased to an average of 1.3 +/- 0.1 ng/ml (p less than 0.05) during the first 60 min after NAL treatment. Serum prolactin (PRL) concentrations for L gilts averaged 4.8 +/- 0.2 ng/ml during the pretreatment period and increased to an average of 6.3 +/- 0.3 ng/ml (p less than 0.05) during the first 60 min after NAL treatment. Serum PRL concentrations averaged 8.6 +/- 0.7 ng/ml and 7.6 +/- 0.6 ng/ml in EF and LF gilts, respectively, prior to NAL treatment, and decreased (p less than 0.05) to an average of 4.1 +/- 0.2 ng/ml and 5.6 +/- 0.4 ng/ml in EF and LF gilts, respectively, during the fourth h after NAL. Naloxone treatment failed to alter serum LH concentrations in EF, LF, or OVX gilts and PRL concentrations in OVX gilts.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of rate of change of luteinizing hormone concentration on in-vitro progesterone secretion within rat corpora lutea during differentiation.

Immature rats were injected with pregnant mares' serum gonadotrophin followed by human chorionic gonadotrophin (hCG). Ovaries were removed 0, 2, 5 or 8 days after hCG and either prepared for morphometric analysis or perifused with 0, 5 or 30 ng luteinizing hormone (LH)/min. In a second study, ovaries were removed on Day 2 or 8 and perifused with 0.1 mg 8-br-cyclic adenosine 5'-phosphate/ml (8-br-cAMP). On Day 0, the granulosa cells of the preovulatory follicles were small (53 +/- 0.5 microns2) with a cytoplasmic to nuclear (Cy:Nu) ratio less than or equal to 1.5. By Day 2, corpora lutea (CL) were present and composed of 95% small luteal cells (diameter less than 125 microns2, Cy:Nu greater than or equal to 3.0) and 5% large luteal cells (diameter greater than 125 microns2, Cy:Nu ratio greater than or equal to 3.0). The percentage of large luteal cells increased to 36 +/- 7% by Day 5, suggesting that they are derived from a select population of small luteal cells. Basal progesterone secretion increased from 38 +/- 5 on Day 0 to 1010 +/- 48 pg/mg/ml on Day 8. The rate of 5 ng LH/min stimulated progesterone secretion on Days 0, 2 and 8; 30 ng LH/min stimulated progesterone secretion on Days 0, 2 and 8, but not on Day 5; 8-br-cAMP stimulated progesterone secretion on both Days 2 and 8. These data demonstrate that once granulosa cells are induced to luteinize they lose their capacity to secrete progesterone in response to 5 ng LH/min and do not regain their responsiveness to LH rate until they completely differentiate. The loss of this LH responsiveness appears to be due to an inability to stimulate sufficient intracellular cAMP concentrations, since cAMP stimulates progesterone secretion on both Days 2 and 8.     

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.     

Pituitary receptors for gonadotropin-releasing hormone in relation to changes in pituitary and plasma luteinizing hormone in ovariectomized-hypothalamo pituitary disconnected ewes. I. Effect of changing frequency of gonadotropin-releasing hormone pulses

Studies were undertaken to determine if changes in the amplitude of luteinizing hormone (LH) pulses that occur in response to changes in the frequency of gonadotropin-releasing hormone (GnRH) pulses are due to an alteration in the number of GnRH receptors. Ewes were ovariectomized (OVX) and the hypothalamus was disconnected from the pituitary (HPD). Ewes were then given pulses of GnRH at a frequency of 1/h or 1/3 h. Two control groups were included: OVX ewes not subjected to HPD, and HPD ewes that were not OVX. At the end of one week of treatment, blood samples were collected to determine the amplitude of LH pulses. The treated ewes were killed just before the next scheduled pulse of GnRH, and the content of LH and number of GnRH receptors were measured in each pituitary. The amplitude of LH pulses was highly correlated with the amount of LH in the pituitary gland (r = 0.71, p less than 0.01), and both LH content and pulse amplitude (mean + SEM) were higher in ewes receiving GnRH once per 3 h (189.7 +/- 39.3 microgram/pituitary, 10.3 +/- 1.1 ng/ml, respectively) than in ewes receiving GnRH once per h (77.8 +/- 11.4 microgram/pituitary, 5.2 +/- 1.3 ng/ml). The pituitary content of LH was highest in the OVX ewes (260.2 +/- 57.4 micrograms/pituitary) and lowest in the nonpulsed HPD ewes (61.7 +/- 51.2 micrograms/pituitary). The number of GnRH receptors was similar in all groups, and was not correlated with any other variable.(ABSTRACT TRUNCATED AT 250 WORDS)     

Luteinizing hormone secretion as influenced by age and estradiol in the prepubertal gilt

The aim of this study was to determine if there is an age related reduction in the sensitivity of the negative feedback action of 17β-estradiol (estradiol) on luteinizing hormone (LH) secretion in the prepubertal gilt. Ovariectomized gilts at 90 (n=12), 150 (n=11) or 210 (n=12) days of age received estradiol benzoate (EB) osmotic pump implants 6/group and the remaining animals received vehicle control (C) implants except for 150-day C (n=5) on Day 0. On Day 10 blood samples were collected every 15 min for 8h and serum LH and estradiol concentrations were measured. Serum estradiol concentrations averaged 5 ± 1, 5 ± 1 and 7 ± 2 pg/ml for the 90-, 150- and 210-day-old gilts implanted with estradiol, respectively, whereas, serum estradiol concentrations was undetectable in C gilts. Mean serum LH concentrations, basal LH concentrations and serum LH pulse amplitude were less in EB-treated gilts at all ages compared to control animals. In contrast, LH pulse frequency initially was less in EB-treated gilts but subsequently increased (P<0.04) with age (from 0.8 ± 0.2 at 90 days to 5.2 ± 0.2/8h at 210 days), and at 210 days of age the pulse frequency was similar to C gilts. These results demonstrate an age related reduction in the sensitivity to the negative feedback action of estradiol on LH secretion and support the idea that the gilt conforms to the gonadostat hypothesis.     

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.     

Concentrations of plasma melatonin and luteinizing hormone in domestic gilts reared under artificial long or short days.

Plasma melatonin concentrations were measured every 1-2 h over 24 h and plasma luteinizing hormone (LH) concentrations every 15 min over 12 h in domestic gilts reared under artificial light regimens that had previously been used to demonstrate photoperiodic effects on puberty. In Expt 1, the light regimens both commenced at 12 h light: 12 h dark (12L:12D) and either increased (long-day) or decreased (short-day) by 15 min/week until the long-day gilts were receiving 16L:8D and the short-day gilts 8L:16D at sampling. In Expt 2, both light regimens commenced at 12L:12D and either increased (long-day) or decreased (short-day) by 10 or 15 min/week to a maximum of 14.5L:9.5D or a minimum of 9.5L:14.5D before being reversed. Sampling took place when daylength had returned to 14L:10D (long-day) or 10L:14D (short-day). In immature gilts housed at 12L:12D (Expt 1) and in postpubertal (Expt 1) and prepubertal (Expt 2) gilts reared under long-day or short-day light regimens, mean plasma melatonin concentrations were basal (3.6 pg/ml) when the lights were on and increased to peak concentrations greater than 15 pg/ml within 1-2 h after dark, before declining gradually to basal concentrations at or near the end of the dark phase. In prepubertal gilts bearing subcutaneous melatonin implants and reared under long-days (Expt 2), mean plasma melatonin concentration in the 6 h before dark was 91.9 +/- 5.26 pg/ml and 125.0 +/- 6.66 pg/ml 1 h after dark, but this increase was not statistically significant. In Expt 2, the short-day gilts had fewer LH pulses (2.6 +/- 0.25 vs. 4.6 +/- 0.24; P less than 0.01) in the 12-h sampling period than the long-day gilts, but the amplitude of the pulses (2.28 +/- 0.23 vs. 1.26 +/- 0.16 ng/ml; P less than 0.01) and the area under the LH curve (78.8 +/- 5.60 vs. 47.3 +/- 6.16; P less than 0.01) was greater in the short-day gilts. In the short-day, but not in the long-day, gilts LH pulses were more frequent (2.0 +/- 0.0 vs. 0.6 +/- 0.25; P less than 0.01), but had a smaller area (61.9 +/- 7.2 vs. 120.2 +/- 23.6; P less than 0.05) in the 6 h of dark than in the 6 h of light, which together made up the 12-h sampling period.(ABSTRACT TRUNCATED AT 400 WORDS)     

Effects of gonadotropin-releasing hormone pulse-frequency modulation on luteinizing hormone, follicle-stimulating hormone and testosterone secretion in hypothalamo/pituitary-disconnected rams

The effects of changes in pulse frequency of exogenously infused gonadotropin-releasing hormone (GnRH) were investigated in 6 adult surgically hypothalamo/pituitary-disconnected (HPD) gonadal-intact rams. Ten-minute sampling in 16 normal animals prior to HPD showed endogenous luteinizing hormone (LH) pulses occurring every 2.3 h with a mean pulse amplitude of 1.11 +/- 0.06 (SEM) ng/ml. Mean testosterone and follicle-stimulating hormone (FSH) concentrations were 3.0 +/- 0.14 ng/ml and 0.85 +/- 0.10 ng/ml, respectively. Before HPD, increasing single doses of GnRH (50-500 ng) elicited a dose-dependent rise of LH, 50 ng producing a response of similar amplitude to those of spontaneous LH pulses. The effects of varying the pulse frequency of a 100-ng GnRH dose weekly was investigated in 6 HPD animals; the pulse intervals explored were those at 1, 2, and 4 h. The pulsatile GnRH treatment was commenced 2-6 days after HPD when plasma testosterone concentrations were in the castrate range (less than 0.5 ng/ml) in all animals. Pulsatile LH and testosterone secretion was reestablished in all animals in the first 7 days by 2-h GnRH pulses, but the maximal pulse amplitudes of both hormones were only 50 and 62%, respectively, of endogenous pulses in the pre-HPD state. The plasma FSH pattern was nonpulsatile and FSH concentrations gradually increased in the first 7 days, although not to the pre-HPD range. Increasing GnRH pulse frequency from 2- to 1-hour immediately increased the LH baseline and pulse amplitude. As testosterone concentrations increased, the LH responses declined in a reciprocal fashion between Days 2 and 7. FSH concentration decreased gradually over the 7 days at the 1-h pulse frequency. Slowing the GnRH pulse to a 4-h frequency produced a progressive fall in testosterone concentrations, even though LH baselines were unchanged and LH pulse amplitudes increased transiently. FSH concentrations were unaltered during the 4-h regime. These results show that 1) the pulsatile pattern of LH and testosterone secretion in HPD rams can be reestablished by exogenous GnRH, 2) the magnitude of LH, FSH, and testosterone secretion were not fully restored to pre-HPD levels by the GnRH dose of 100 ng per pulse, and 3) changes in GnRH pulse frequency alone can influence both gonadotropin and testosterone secretion in the HPD model.     

Modulation of luteinizing hormone pulse amplitude by the frequency of luteinizing hormone-releasing hormone stimulation and by testosterone in castrated, hypothalamic-lesioned male rats

The frequency of spontaneous luteinizing hormone (LH) pulses is thought to be a direct result of the frequency of luteinizing hormone-releasing hormone (LHRH) pulses from the hypothalamus. By contrast, the amplitude of spontaneous LH pulses may be controlled by several factors other than the amplitude of LHRH pulses. We tested two hypotheses: 1) that LH pulse amplitude is determined in part by the frequency of LHRH pulses of constant magnitude, and 2) that testosterone (T) exerts a direct feedback effect on the pituitary gland to regulate LH pulse amplitude. Gonadal feedback was eliminated by castrating adult male rats (n = 20). Endogenous LHRH secretion was eliminated by lesioning the medial basal hypothalamus. Serum LH levels (0.19 +/- 0.04 ng/ml RP-2, mean +/- SEM) and T levels (0.15 +/- 0.02 ng/ml), measured several weeks after hypothalamic lesioning, confirmed the hypogonadotropic hypogonadal state of the animals. During a 8-h period, unanesthetized, unrestrained animals were injected with 40-ng pulses of LHRH via catheters into the jugular vein, and blood samples for LH measurement were drawn at 10-min intervals. The LHRH pulse interval was 20 min during the first 4 h in all animals. The pulse interval was doubled to 40 min in half of the animals (n = 10) during the next 4 hours; in the other 10 animals, the pulse interval was maintained constant at 20 min throughout the study. Within both of these groups, one-half of the animals (n = 5) were infused with T to achieve a physiological level of T in serum (2.46 +/- 0.36 ng/ml at 4 h), while the other half received vehicle.(ABSTRACT TRUNCATED AT 250 WORDS)     

Pattern of ovarian progesterone secretion during the luteal phase of the ovine estrous cycle

Pulsatile secretion of progesterone has been observed during the late luteal phase of the menstrual cycle in the rhesus monkey and human. As the luteal phase progresses in each of these species, there is a pattern of decreased frequency and increased amplitude of progesterone pulses. The present study was designed to determine the pattern of progesterone secretion during the late luteal phase (Days 10-16) of the normal ovine estrous cycle. Five unanesthetized ewes, each bearing an indwelling cannula in the utero-ovarian vein, were bled every 15 min from 0800 h on Day 10 through 0800 h on Day 16 of the estrous cycle. With the computer program PULSAR, it was determined that progesterone secretion was episodic, with pulsations observed on all days. Analysis of variance was used to determine differences in frequency, amplitude, and interpeak interval (IPI) of progesterone pulses among ewes and days. The ewes averaged 8.0 +/- 0.63 pulses of progesterone per 24 h. Mean frequency of pulses was not different between days but showed differences between ewes. Mean amplitude of progesterone pulses was 7.0 +/- 0.27 ng/ml, with no differences observed either between days or between ewes. Mean IPI was 197 +/- 7.1 min, and, like frequency, the IPI was not different between days, but varied between ewes. No consistent temporal relationship was found between progesterone pulses and luteinizing hormone (LH), as determined by bioassay and radioimmunoassay, on Day 14 of the cycle in one ewe. The results indicate that progesterone secretion is episodic during the luteal phase of the ovine estrous cycle and is independent of LH pulses.(ABSTRACT TRUNCATED AT 250 WORDS)     

Adenylate cyclase activity of the corpus luteum during the oestrous cycle of the pig

Basal adenylate cyclase values for corpora lutea (CL) removed from cyclic gilts on Days 3, 8, 13 and 18 were 178 +/- 61, 450 +/- 46, 220 +/- 25 and 208 +/- 18 pmol cAMP formed/min/mg protein, respectively. Basal activity was significantly elevated on Day 8 (P less than 0.001). LH-stimulatable adenylate cyclase values for CL from Days 3, 8, 13 and 18 were 242 +/- 83, 598 +/- 84, 261 +/- 27 and 205 +/- 17 pmol cAMP formed/min/mg protein respectively. Serum progesterone concentrations of 12 gilts bled every 2 days through one complete oestrous cycle ranged from 1.1 to 26.9 ng/ml with highest values between Days 8 and 12. The decline in serum progesterone concentrations was coincident with the decrease in basal adenylate cyclase activity. There was no LH-stimulatable adenylate cyclase activity present in the CL at the specific times of the oestrous cycle examined. We conclude that progesterone secretion by the pig CL is apparently dependent on basal activity of adenylate cyclase.     

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)     

Changing frequency of pulsatile luteinizing hormone and progesterone secretion during the luteal phase of the menstrual cycle of rhesus monkeys

Experiments were conducted to examine the pulsatile nature of biologically active luteinizing hormone (LH) and progesterone secretion during the luteal phase of the menstrual cycle in rhesus monkeys. As the luteal phase progressed, the pulse frequency of LH release decreased dramatically from a high of one pulse every 90 min during the early luteal phase to a low of one pulse every 7-8 h during the late luteal phase. As the pulse frequency decreased, there was a corresponding increase in pulse amplitude. During the early luteal phase, progesterone secretion was not episodic and there were increments in LH that were not associated with elevations in progesterone. However, during the mid-late luteal phase, progesterone was secreted in a pulsatile fashion. During the midluteal phase (Days 6-7 post-LH surge), 67% of the LH pulses were associated with progesterone pulses, and by the late luteal phase (Days 10-11 post-LH surge), every LH pulse was accompanied by a dramatic and sustained release of progesterone. During the late luteal phase, when the LH profile was characterized by low-frequency, high-amplitude pulses, progesterone levels often rose from less than 1 ng/ml to greater than 9 ng/ml and returned to baseline within a 3-h period. Thus, a single daily progesterone determination is unlikely to be an accurate indicator of luteal function. These results suggest that the changing pattern of mean LH concentrations during the luteal phase occurs as a result of changes in frequency and amplitude of LH release. These changes in the pulsatile pattern of LH secretion appear to have profound effects on secretion of progesterone by the corpus luteum, especially during the mid-late luteal phase when the patterns of LH concentrations are correlated with those of progesterone.     

Changes in patterns of luteinizing hormone secretion before and after the first ovulation in the postpartum mare

To determine whether luteinizing hormone (LH) secretion during the first estrous cycle postpartum is characterized by pulsatile release, circulating LH concentrations were measured in 8 postpartum mares, 4 of which had been treated with 150 mg progesterone and 10 mg estradiol daily for 20 days after foaling to delay ovulation. Blood samples were collected every 15 min for 8 h on 4 occasions: 3 times during the follicular phase (Days 2-4, 5-7, and 8-11 after either foaling or end of steroid treatment), and once during the luteal phase (Days 5-8 after ovulation). Ovulation occurred in 4 mares 13.2 +/- 0.6 days postpartum and in 3 of 4 mares 12.0 +/- 1.1 days post-treatment. Before ovulation, low-amplitude LH pulses (approximately 1 ng/ml) were observed in 3 mares; such LH pulses occurred irregularly (1-2/8 h) and were unrelated to mean circulating LH levels, which gradually increased from less than 1 ng/ml at foaling or end of steroid treatment to maximum levels (12.3 ng/ml) within 48 h after ovulation. In contrast, 1-3 high-amplitude LH pulses (3.7 +/- 0.7 ng/ml) were observed in 6 of 7 mares during an 8-h period of the luteal phase. The results suggest that in postpartum mares LH release is pulsatile during the luteal phase of the estrous cycle, whereas before ovulation LH pulses cannot be readily identified.