Opioid inhibition of luteinizing hormone secretion during the postpartum period in suckled beef cows

Recent studies have shown that naloxone (N), an opioid antagonist, increases concentrations of luteinizing hormone (LH) in the postpartum anestrous beef cow. However, the LH response to N was influenced by the postpartum interval. For example, a significant LH response to 200 mg of N occurred on d 42 but not on d 14 or 28 postpartum. The present study was conducted to determine the effect of different doses of N on LH secretion during the postpartum period of beef cows. Twelve cows were given 200, 400 or 800 mg of N on d 14, 28 and 42 postpartum in a Latin square design with repeat measures within cells. On d 14, serum concentrations of LH increased (P less than .01) from .5 +/- .1 ng/ml (mean +/- SE) before N to a peak of 2.0 +/- .5 and 1.4 +/- .5 ng/ml for cows given 400 and 800 mg of N, respectively. In contrast, 200 mg of N had no effect on serum concentrations of LH. On d 28 and 42 all three doses of N elevated (P less than .01) serum concentrations of LH. Therefore, a larger dose of N was required to increase serum concentrations of LH on d 14 postpartum compared with d 28 and 42. Based on these data we suggest that endogenous opioids participate in the regulation of LH secretion in the early postpartum period. The differential response to naloxone may be due to changes in endogenous opioid inhibition of LH secretion during the postpartum period.     

Neuroendocrine regulation of growth hormone secretion in sheep. V. Growth hormone releasing factor and thyrotrophin releasing hormone.

The effects of intravenous (IV) and intracerebroventricular (ICV) administration of either bovine growth hormone releasing hormone (GRF) or thyrotrophin releasing hormone (TRH) on plasma growth hormone (GH) and glucose levels have been examined in sheep. Intravenous GRF 1-29NH2 at 3 and 30 micrograms stimulated an increase in GH levels in a dose-dependent fashion; administration of GRF into a lateral cerebral ventricle, however, produced a smaller GH response which was similar at these two doses. Evaluation of somatostatin levels in petrosal sinus blood (which collects pituitary effluent blood) showed that ICV administration of GRF stimulated a release of somatostatin into the blood. Furthermore, concurrent administration of GRF and a potent anti-somatostatin serum ICV resulted in a much enhanced release of GH which was similar to that obtained with a comparable dose of GRF given IV. TRH (as another putative GH-secretagogue) was also administered both IV and ICV. When given IV, 200 micrograms (but not 100 micrograms) TRH produced an elevation in GH levels. By contrast, when 5 micrograms TRH was given ICV there was a decrease in circulating GH levels, but no change in plasma somatostatin concentrations. These results indicate that the smaller GH response to ICV- compared with IV-administered GRF is due to the release of somatostatin within the brain. In addition, it would seem that TRH is not a physiological GH-secretagogue in sheep.     

Plasma somatotropin response to exogenous growth hormone releasing factor in lambs

Two experiments were performed to examine the ability of human pancreatic growth hormone releasing factor (hGRF) administration to stimulate endogenous growth hormone (GH) secretion in lambs. Each study utilized eight Dorset wether lambs in replicated 4 X 4 Latin square experiments. Growth hormone response (integrated area under the curve for 150 min post-injection) for 0, 1, 5 and 10 micrograms hGRF/kg body weight averaged 13, 23, 92 and 134 units, respectively. While the 1-microgram hGRF dose was not different (P greater than .05) than the response to saline injection, there was an increased (P less than .01) GH response to 5 or 10 micrograms hGRF. Overall the GH response increased in a log dose-response fashion. There was distinct variation between lambs in their response to hGRF. Study II examined the optimal method to administer 40 micrograms hGRF/kg body weight to maximize GH concentration over 24 h. Continuous infusion (CI) was compared with eight (8X), four (4X), or two (2X) injections/d. Hourly blood samples were obtained from all lambs. Growth hormone response (area under the curve for 24 h) was 162, 305, 306 and 220 units for CI, 8X, 4X and 2X, respectively. Growth hormone response to CI was inferior to discrete injections, and the GH response to 4X or 8X was superior to 2X/d. Results demonstrate that, in spite of lamb-to-lamb variation, one can utilize exogenous hGRF to enhance GH secretion in lambs. Thus, the ability of exogenous hGRF to enhance growth performance merits further study.     

Ovarian compensatory hypertrophy following unilateral ovariectomy in the suckled sow

The effects of unilateral ovariectomy on ovarian compensatory hypertrophy (OCH), endocrine profiles and the pituitary response to gonadotropin releasing hormone (GnRH) were studied in 46 multiparous suckled sows. On d 20 of lactation (d 0 of experiment), sows were subjected to sham ovariectomy (Sham; n = 23) or unilateral ovariectomy (ULO; n = 23). On d 1 (n = 16), 2 (n = 15) or 8 (n = 15) following initial surgery the remaining ovaries in both Sham and ULO sows were removed. Immediately following removal of the remaining ovaries, GnRH (10 micrograms) was administered to each sow. Peripheral blood samples were taken every 10 min for 80 min beginning 20 min prior to GnRH administration. No difference in ovarian weight was observed between ULO and Sham sows until d 8, when ovarian weight was greater (P less than .05) for the remaining ovary from ULO sows (3.96 +/- .21 vs 5.91 +/- .39 g). Ovarian follicular fluid weights from ULO sows were greater (P less than .05) than Sham sows on both d 2 and 8. On d 1, plasma concentrations of follicle stimulating hormone (FSH) were greater (P less than .05) in ULO sows than in Sham sows (2.9 +/- .2 vs 2.1 +/- .1 ng/ml). Plasma FSH concentrations, however, did not differ between Sham and ULO sows on either d 2 or 8. Ovarian venous concentrations of estradiol-17 beta were also greater (P less than .05) in ULO sows compared with Sham sows on d 2 but not d 8.     

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.     

Opioid modulation of gonadotropin releasing hormone release from the hypothalamic preoptic area in the pig

Two experiments (Exp) were conducted to examine in vitro the release of gonadotropin releasing hormone (GnRH) from the hypothalamus after treatment with naloxone (NAL) or morphine (MOR). In Exp 1, hypothalamic-preoptic area (HYP-POA) collected from 3 market weight gilts at sacrifice and sagitally halved were perifused for 90 min prior to a 10 min pulse of morphine (MOR; 4.5 × 10⁻⁶ M) followed by NAL (3.1 × 10⁻⁵ M) during the last 5 min of MOR (MOR + NAL; N=3). The other half of the explants (n=3) were exposed to NAL for 5 min. Fragments were exposed to KCl (60 mM) at 175 min to assess residual GnRH releasability. In Exp 2, nine gilts were ovariectomized and received either oil vehicle im (V; n=3); 10 μg estradiol-17β/kg BW im 42 hr before sacrifice (E; n=3); .85 mg progesterone/kg BW im twice daily for 6 d prior to sacrifice (P₄; n=3). Blood was collected to assess pituitary sensitivity to GnRH (.2 μg/kg BW) on the day prior to sacrifice. On the day of sacrifice HYP-POA explants were collected and treated as described in Exp 1 except tissue received only NAL. In Exp 1, NAL increased (P<.05) GnRH release. This response to NAL was attenuated (P<.05) by coadministration of MOR. Cumulative GnRH release after NAL was greater (P<.05) than after MOR + NAL. All tissues responded similarly to KCl with an increase (P<.05) in GnRH release. In Exp 2, pretreatment luteinizing hormone (LH) concentrations were lower (P<.05) in E gilts compared to V and P₄ animals with P₄ being lower (P<.05) than V gilts. LH response to GnRH was lower (P<.05) in E pigs than in V and P₄ animals, while the responses was similar between V and P₄ gilts. NAL increased GnRH release in all explants, whereas, KCl increased GnRH release in 6 of 9 explants. These results indicate that endogenous opioid peptides may modulate in vitro GnRH release from the hypothalamus in the gilt.     

猪生产中GRF基因质粒的应用研究进展

随着分子生物学的发展,将生长激素释放因子(GRF)基因质粒注入到动物体内,并改变动物机体的生长机能和营养调控已成为现代营养学研究的热点。本文主要从pGRF的来源、作用特点、作用机理及在猪生产中的应用等方面进行了概述。     

Thyrotropin releasing hormone interactions with growth hormone secretion in horses

Light horse mares, stallions, and geldings were used to 1) extend our observations on the thyrotropin releasing hormone (TRH) inhibition of GH secretion in response to physiologic stimuli and 2) test the hypothesis that stimulation of endogenous TRH would decrease the normal rate of GH secretion. In Exp. 1 and 2, pretreatment of mares with TRH (10 microg/kg BW) decreased (P < 0.001) the GH response to exercise and aspartate infusion. Time analysis in Exp. 3 indicated that the TRH inhibition lasted at least 60 min but was absent by 120 min. Administration of a single injection of TRH to stallions in Exp. 4 increased (P < 0.001) prolactin concentrations as expected but had no effect (P > 0.10) on GH concentrations. Similarly, 11 hourly injections of TRH administered to geldings in Exp. 5 did not alter (P > 0.10) GH concentrations either during the injections or for the next 14 h. In Exp. 5, it was noted that the prolactin and thyroid-stimulating hormone responses to TRH were great (P < 0.001) for the first injection, but subsequent injections had little to no stimulatory effect. Thus, Exp. 6 was designed to determine whether the inhibitory effect of TRH also waned after multiple injections. Geldings pretreated with five hourly injections of TRH had an exercise-induced GH response identical to that of control geldings, indicating that the inhibitory effect was absent after five TRH injections. Retrospective analysis of pooled, selected data from Exp. 4, 5, and 6 indicated that endogenous GH concentrations were in fact lower (P < 0.01) from 45 to 75 min after TRH injection but not thereafter. In Exp. 7, 6-n-propyl-2-thiouracil was fed to stallions to reduce thyroid activity and hence thyroid hormone feedback, potentially increasing endogenous TRH secretion. Treated stallions had decreased (P < 0.01) concentrations of thyroxine and elevated (P < 0.01) concentrations of thyroid-stimulating hormone by d 52 of feeding, but plasma concentrations of GH and prolactin were unaffected (P > 0.10). In contrast, the GH response to aspartate and the prolactin response to sulpiride were greater (P < 0.05) in treated stallions than in controls. In summary, TRH inhibited exercise- and aspartate-induced GH secretion. The duration of the inhibition was at least 1 h but less than 2 h, and it waned with multiple injections. There is likely a TRH inhibition of endogenous GH episodes as well. Reduced thyroid feedback on the hypothalamic-pituitary axis did not alter basal GH and prolactin secretion.     

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).     

Effect of active immunization against growth hormone releasing factor on concentrations of somatotropin and insulin-like growth factor I in lactating beef cows.

Two experiments were conducted to determine the effects of immunoneutralization of growth hormone-releasing factor [GRF(1-29)-NH2] on concentrations of somatotropin (ST) and insulin-like growth factor I (IGF-I) in lactating beef cows. In Experiment 1, multiparous Hereford cows were immunized against 2 mg GRF(1-29)-(Gly)4-Cys-NH2 conjugated to human serum albumin (GRFi, n = 3) or 2 mg human serum albumin (HSAi, n = 3) at 52 +/- 1 d prior to parturition. Boosters (1 mg) were administered on days 12, 40 and 114 postpartum (pp). Serum samples were collected at 15-min intervals for 5 hr on days 18, 46 and 120 pp, followed by administration (IV) of an opioid agonist (FK33-824; 10 micrograms/kg) and an antagonist (naloxone; .5 mg/kg) at hours 5 and 7, respectively. A GRF-analog ([desamino-Tyr1, D-Ala2, Ala15] GRF (1-29)-NH2; 3.5 micrograms/kg) and arginine (.5 g/kg) were administered at hour 10 on days 47 and 121, respectively. Percentage binding of [125I]GRF (1:100 dilution of serum) 28 d after primary immunization was greater in GRFi (14.3 +/- 4.9) than in HSAi (.7 +/- .3) cows. Binding increased to 29.3 +/- 6.5% after first booster in GRFi cows. Episodic release of ST was abolished by immunization against GRF; concentration and frequency of release of ST were lower (P less than .05) in GRFi than in HSAi cows on all days pp. Concentrations of IGF-I were lower in GRFi than in HSAi cows throughout lactation. Serum ST failed to increase following FK33-824 or arginine in GRFi; however, ST increased after both compounds in HSAi cows. Concentrations of ST following GRF-analog were greater (P less than .05) in HSAi than in GRFi cows. Experiment 2 was conducted to determine if a lower dose of antigen and a single booster would be sufficient to lower ST and IGF-I in lactating cows. Multiparous Hereford and Angus cows were assigned to GRFi (n = 6) or HSAi (n = 6). Primary (1.2 mg) and booster (.5 mg) immunizations were administered -14 and 8 d from calving, respectively. Cows were restricted to 60% of recommended intake of energy during lactation in order to elevate concentrations of ST. Serum samples were collected at 15-min intervals for 6 hr on days 26, 50, 73, 90 and 109 pp. Two of six GRFi cows had binding less than 10% (1:1,000 dilution of serum) and were omitted from further analyses.(ABSTRACT TRUNCATED AT 400 WORDS)     

Regulation of growth hormone expression by thyrotropin-releasing hormone through the pituitary-specific transcription factor Pit-1 in chicken pituitary

Pit-1 is a pituitary-specific POU-domain DNA binding factor, which binds to and trans-activates promoters of growth hormone- (GH), prolactin- (PRL) and thyroid stimulating hormone beta- (TSHbeta) encoding genes. Pit-1 has been identified in several mammalian and avian species. Thyrotropin-releasing hormone (TRH) is located in the hypothalamus and it stimulates TSH, GH and PRL release from the pituitary gland. In the present study, we successfully developed a competitive RT-PCR for the detection of Pit-1 expression in the chicken pituitary, that was sensitive enough to detect picogram levels of Pit-1 mRNA. Applying this method, the effect of TRH injections on Pit-1 mRNA expression was determined in the pituitary of chick embryos and growing chicks. In both 18-day-old embryos and 10-day-old male chicks the Pit-1 mRNA expression was significantly increased following TRH injection, thereby indicating that the stimulatory effects of TRH on several pituitary hormones is mediated via its effect on Pit-1 expression. Therefore, a semi-quantitative RT-PCR method was used to detect possible changes in GH levels. TRH affected the GH mRNA levels at both developmental stages. These results, combined with the data on Pit-1 mRNA expression, indicate that Pit-1 has a role in mediating the stimulatory effects of TRH on pituitary hormones like GH.     

Effects of plasmid-mediated growth hormone releasing hormone supplementation in young,healthy Beagle dogs

Our study focused on the evaluation of the pharmacological and toxicological effects of plasmid-mediated GHRH supplementation with electroporation in normal adult dogs over a 180-d period. Twenty-eight dogs (< 2 yr of age) were randomized to four groups. Three groups (four dogs/sex for each group) were treated with ascending doses of GHRH-expressing plasmid: 0.2, 0.6, and 1 mg. One group (two dogs of each sex) served as the control. Clinical observations and body weights were recorded. Hematological, serum biochemical, and urine analyses were performed. Serum IGF-I, ACTH, and insulin were determined. Necropsies were performed on d 93 and 180; organs were weighed and tissues were fixed and processed for light microscopy. Selected tissues were used to assess plasmid biodistribution on d 93. At all doses, plasmid GHRH caused increased weight gain (P < 0.001), without organomegaly. Serum glucose and insulin in fasted dogs remained within normal ranges at all time points. Adrenocorticotropic hormone was normal in all groups. Significant increases in number of red blood cells, hematocrit, and hemoglobin (P < 0.01) were observed. In conclusion, our study shows that plasmid-mediated GHRH supplementation is safe in electroporated doses up to 1.0 mg in young healthy dogs.     

Potent interaction between thyrotropin releasing hormone (TRH) and human pancreatic growth hormone releasing factor (hpGRF) in stimulating chicken growth hormone (cGH) in vivo: Hypothalamic noradrenergic mediation in TRH stimulation of cGH release

The interaction of human pancreatic growth hormone releasing factor (hpGRF) and thyrotropin releasing hormone (TRH) on chicken growth hormone (cGH) release in vivo and possible noradrenergic involvement on TRH-induced stimulation of cGH in vivo were examined. Four-week old cockerels (1 kg) were injected intravenously with hpGRF (1.0 μg/bird), TRH (0.1 μg/bird), or hpGRF (1.0 μg/bird) in combination with TRH (0.1 μg/bird). Five min after the injection, blood samples were collected and serum concentrations of cGH were determined by a homologous RIA. The results showed that hpGRF and TRH were potent stimulators of cGH release, 5- and 6-fold over the control birds, respectively, and that hpGRF and TRH administered in combination produced a synergistic stimulation of cGH release (>20 fold). In separate experiments, pretreatment with alpha-methyl-para-tyrosine (250 mg/bird) for 2 hours resulted in complete suppression of the TRH stimulatory effect on cGH release but not the stimulatory effect of hpGRF. Pretreated with phenoxybenzamine hydrochloride (20 mg/bird) or diethyl-dithiocarbamate (500 mg/bird) also resulted in complete suppression of TRH-induced cGH release. These results indicate that hpGRF acts directly at the pituitary and TRH acts at the hypothalamus in addition to the pituitary in stimulating cGH release, possibly mediated through the noradrenergic neurons. HpGRF and TRH were potent releasers of cGH and their stimulation was potentiated when administered together.     

Steroid hormone and luteinizing hormone concentrations in the anestrous sow.

This investigation characterized serum concentrations of luteinizing hormone (LH), estradiol-17 beta (E2), progesterone (P4) and cortisol (C) in anestrous sows. Twenty-two sows that had not returned to estrus within 45 days after weaning (anestrous sows), and ten sows that had returned to estrus within seven days following weaning (cyclic sows) were nonsurgically fitted with indwelling jugular vein cannulae. Blood samples were collected at 6 h intervals for seven days and at 15 min intervals for 8 h on the fifth day after cannulation. Serum LH concentrations were determined in all samples, while C, E2 and P4 levels were quantitated in serum collected at 6 h intervals. Serum P4 concentrations in anestrous sows were consistently less than 0.5 ng/mL, and E2 levels ranged from 10 to 19 pg/mL. Concentrations of LH remained less than 1.0 ng/mL in anestrous sows, whereas a preovulatory LH surge was observed in five of ten cyclic sows. There was a circadian rhythm in mean C levels with C peaks occurring at 0600 or 2400 h and nadir levels observed at 1200 and 1800 h. Few differences in C levels were detected between anestrous and cyclic sows. It was evident that anestrous sows did not exhibit cyclic or predictable variations in steroid hormone concentrations. Unfortunately, the results of this study failed to elucidate the endocrine pathogenesis of the anestrous sow.     

Growth hormone releasing factors and secretion of growth hormone in sheep,calves and pigs

Human pancreatic growth hormone releasing factors (hpGRF) (1–40) and (1–44) were administered iv in sheep, pigs and calves to determine their effectiveness in stimulating GH release in these species. Both peptides produced a rapid increase in plasma GH concentration in all three species at dose levels ranging from .0065 to .65 nmol/kg. Moreover, there was no difference in the GH-secretory response observed between hpGRF(1–44)NH₂ and (1–40)OH in sheep. Sheep also responded to hpGRF(1–40)NH₂ and (1–40)OH as well as [his₁]- and [tyr₁]-hpGRF(1–40)NH₂ in a similar manner. Rat hypothalamic GRF was less effective than [his₁]-hpGRF(1–40), while the response to bGRF was not significantly different from hpGRF(1–40) in stimulating GH secretion in sheep. Although all three species responded to hpGRF, the elevation in plasma GH levels above baseline were greater after hpGRF injection in sheep than in pigs or calves. Subcutaneous injection of hpGRF in sheep was an effective mode of administration of the peptide, although the effect was not as long-lasting as that after iv injections and higher doses were required to stimulate GH secretion.     

Injection of porcine growth hormone releasing hormone gene plasmid in skeletal muscle increases piglets' growth and whole body protein turnover

The aim of the current study was to investigate the effects of a porcine growth hormone releasing hormone (pGHRH) gene plasmid injection in piglets on growth performance and whole body protein turnover. Sixty male Canadian Landrace × Chinese Taihu piglets were assigned to an intramuscular injection of 0 (control), 0.25, 0.5, 1 and 2 mg. All pigs were fed with the same diet (crude protein: 239.8 g/kg, digestible energy: 14.28 MJ/kg) at ad libitum intake. Protein turnover was determined on the 22nd day with a three-pool model by using a single-dosage, end-product analysis method with ¹⁵ N-glycine as a tracer. Injection of the pGHRH gene plasmid increased the piglets' growth rate, altered feed intake and decreased feed conversion ratio. It increased plasma growth hormone releasing hormone (GHRH), growth hormone (GH), insulin-like growth factor-I (IGF-I) and somatostatin but reduced serum urea and triglyceride. It reduced the urinary nitrogen excretion and led to higher nitrogen retention as well as the efficiencies of nitrogen retention and digestible N utilization. It increased the rates of protein synthesis, protein breakdown and net protein gain. Excretion of endogenous urinary nitrogen was reduced and nitrogen reutilization rate was improved. Conclusions: Injection of the pGHRH gene plasmid in skeletal muscle stimulated GHRH, GH and IGF-I excretion in piglets. Protein deposition was increased by an increase in protein synthesis and a smaller increase in protein breakdown, which was accompanied by reducing amino acid oxidation and increasing nitrogen reutilization.     

Effects of corticotropin releasing factor on the production of adrenocorticotropic hormone by leukocyte populations

1. Corticotropin-releasing factor (CRF), a neuropeptide with immunomodulating properties, is known to stimulate avian splenic leukocytes to produce adrenocorticotropic hormone (ACTH). 2. The present study was to determine which avian splenic leukocyte subpopulation(s) produce ACTH in response to CRF stimulation. 3. Splenic leukocytes from 8-week-old male chickens were isolated on Histopaque 1077 and macrophages were separated from lymphocytes by adherence to a polystyrene surface. 4. Different concentrations of CRF (0, 5, 50, 500 or 1000 ng/m) were incubated with the different leukocyte populations, supernatants were collected and ACTH was measured using a radioimmunoassay. 5. Isolated macrophages, stimulated with CRF, produced significantly more ACTH than either unstimulated macrophages or CRF-stimulated lymphocytes, suggesting that ACTH may be produced by a particular subset of leukocytes, the macrophages (and monocytes), in response to CRF stimulation.     

Estradiol feedback inhibition of luteinizing hormone concentrations in the anestrous sow

The suppressive effects of exogenous 17 beta-estradiol (E2) on LH concentrations in sows that remained anestrus following weaning and in those that returned to estrus were evaluated. Four anestrous and four cyclic sows were treated subcutaneously with silastic implants containing E2 at 13 d after ovariectomy (d 0). Three anestrous and six cyclic sows received silastic implants without E2. Blood was collected at 6-h intervals from d -1 to d 12 and at 15-min intervals for 8 h on d -1, 2, 7 and 12. Sows were treated with 1 microgram GnRH/kg BW at the completion of each 8-h frequent sampling period. Blood was collected at intervals of 10 to 30 min for 3 h after GnRH treatment. Concentrations of E2 remained less than 5 pg/ml in sham-treated sows and were between 20 and 25 pg/ml in E2-treated females. Pulsatile LH concentrations was similar between anestrous and cyclic sows prior to implant treatment. Sham-treated anestrous sows had greater (P less than .05) pulse frequency and mean LH concentrations than E2-treated anestrous sows on d 2, 7 and 12. Differences in pulsatile LH concentrations between E2-treated and sham-treated cyclic sows were not detected. Pulse frequency was less (P less than .05) in E2-treated anestrous sows than in E2-treated cyclic sows on d 7 and 12. Peak LH concentrations were greater (P less than .05) in E2-treated cyclic sows than in E2-treated anestrous sows at each GnRH challenge. These results suggest that the hypothalamo-hypophyseal axis is more sensitive to the negative feedback effects of E2 in anestrous sows than in cyclic sows. In addition, chronic E2 treatment reduces pituitary responsiveness to GnRH to a greater extent in anestrous than in cyclic sows. Failure to return to estrus in swine may be due, at least in part, to an increased sensitivity of the hypothalamo-hypophyseal axis to the negative feedback effect of estradiol.