Effect of Naloxone on Serum Luteinizing Hormone Concentration in Anovulatory Holstein Cows During the Early Postpartum Period

This experiment was conducted to investigate the effect of naloxone (NAL), an opioid receptor antagonist, on pituitary LH secretion in anovulatory Holstein cows during the early postpartum period when cows were expected to be in negative net energy balance. Twenty-three cows (11 primiparous) received either saline (n = 12) or 1 mg/kg BW NAL i.v. (n = 11) on Day 14 or 15 postpartum. Jugular blood samples were collected at 15-min intervals for 2 hr before and 2.5 hr after NAL or saline. All cows received 3 ug gonadotropin releasing hormone (GnRH) at 2.5 hr post-NAL or -saline and blood collection was continued for 1 hr. Mean serum progesterone concentration was 0.33 ± 0.2 ng/ml. Mean net energy balance for all cows was -5.5 ± 0.6 Mcal/day. Naloxone caused a transient increase (P < 0.05) in serum LH concentrations in both primi- and multiparous cows within 45 min after administration. In contrast, serum LH concentrations remained unchanged in saline-treated cows. GnRH increased (P < 0.05) LH and there was no effect of treatment. These results suggest that modulation of LH secretion, at least in part, may be mediated via endogenous opioids in dairy cows before first postpartum ovulation.     

Interactions of photoperiod, testosterone, and naloxone on GnRH and LH pulse parameters in the male sheep

This study tested the hypothesis that the effects of the opiate antagonist naloxone on GnRH (and LH) secretion is affected by photoperiod length and testosterone (T) concentrations. The effect of infusing naloxone on GnRH and LH pulse patterns was determined in four groups of orchidectomized sheep: long day (LD) photoperiod treated with T, LD without T (LDC), short day photoperiod (SD) with T, SDC (n = 5-7/group). Hypophyseal-portal and jugular blood samples were collected at 10 min intervals for 4 h before and 4 h during naloxone infusion (1 mg/kg/h). Neither photoperiod nor T affected either mean GnRH or LH whereas naloxone (P < 0.01) increased both. LD photoperiod (P < 0.01), T (P < 0.01) and naloxone (P < 0.01) all increased LH pulse amplitude whereas only naloxone increased GnRH pulse amplitude (P < 0.01). There was an interaction (P < 0.01) between steroid and naloxone on LH, but not GnRH, pulse amplitude. Both LD photoperiod and T increased both LH and GnRH (P < 0.01) interpulse-interval (IPI). Naloxone decreased GnRH IPI (P < 0.01). The LH/GnRH pulse amplitude ratio was (P < 0.02) increased by T--likely a secondary response to the T-induced increase in IPI. These results are interpreted as showing that in the ram the endogenous opiate peptides regulate both GnRH pulse frequency and amplitude, but that their specific role is modulated by photoperiod and T. These results do not support the concept that the opiate peptides are the primary mediators of the negative feedback effects of T.     

Influence of calf removal on the serum luteinizing hormone response to naloxone in the postpartum beef cow

Twelve anestrous, postpartum beef cows were used to determine the effect of calf removal on the effect of naloxone on serum luteinizing hormone (LH) concentrations. On d 1, six cows were injected iv with saline and six with 200 mg naloxone dissolved in saline. Blood samples were taken at 15-min intervals for 2 h before and 2 h after naloxone or saline administration. At the beginning of blood sampling, calves were removed from three cows in each treatment. At 48 h after calf removal (d 3), all cows were injected iv with 200 mg naloxone and blood samples were collected as on d 1. On d 1, naloxone treatment increased (P less than .01) serum LH concentrations from 1.2 +/- .3 ng/ml at time 0 to 4.3 +/- .6 ng/ml and 4.7 +/- .8 ng/ml at 15 and 30 min, respectively. Injection of saline had no effect on serum LH concentrations. Forty-eight-hour calf removal increased (P less than .01) serum LH concentrations in five of six cows (1.7 +/- .8 vs 4.4 +/- 1.2 ng/ml). Naloxone treatment failed to increase serum LH concentrations in these cows. Injection of naloxone increased (P less than .01) serum LH concentrations in the one cow that did not exhibit an LH increase after calf removal and in six cows whose calves were not removed (1.4 +/- .2 vs 4.4 +/- .5 ng/ml). The present study provides additional evidence that endogenous opioids regulate LH in the postpartum beef cow. We hypothesize that suckling stimulates an opioid inhibition of LH secretion and removal of the suckling stimulus removes the opioid inhibitory tone.     

Endogenous opioid suppression of release of luteinizing hormone during suckling in postpartum anestrous beef cows

Beef cows were used to determine if suckling influences release of LH via endogenous opioids at 28 +/- 4 d after parturition. Cows of similar weight and body condition (6.8 +/- .1, 1 = emaciated, 9 = obese) were assigned randomly to five groups (n = 6 to 7): 1) control-suckled/saline (suckled 15 min every 6 hr for 48 hr); 2) control-suckled/naloxone; 3) calf-removal/saline (calf removal for 52 hr); 4) calf-removal/naloxone; and 5) control-suckled/GnRH (Gonadotropin-Releasing Hormone). At 0 hr, saline was administered to all cows. This treatment was continued at 6 hr intervals for 24 hr. Either naloxone (0.5 mg/kg), GnRH (40 ng/kg) or saline was administered to cows in their respective groups every 6 hr during the ensuing 24-hr period in calf-removal groups, or immediately preceding each suckling episode in the control-suckled groups. Blood samples for analysis of luteinizing hormone (LH) were collected at 15-min intervals for 1 hr prior to and 3 hr after treatment at 0, 24, 36 and 48 hr. Cows were observed for estrus twice daily. All cows in the control-suckled/GnRH group released LH (P less than .05) in response to exogenous GnRH, indicating the presence of releasable quantities of the gonadotropin. Mean concentrations of LH were not effected (P greater than .05) by the control-suckled regime. However, calf-removal alone, or in combination with naloxone, increased (P less than .05) mean concentrations of LH by 48 hr.(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of dose, frequency, and duration of infused gonadotropin-releasing hormone on secretion of luteinizing hormone and follicle-stimulating hormone in nutritionally anestrous beef cows.

Nutritionally induced anovulatory cows were ovariectomized and used to determine the relationships between dose, frequency, and duration of exogenous gonadotropin-releasing hormone (GnRH) pulses and amplitude, frequency, and concentrations of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in serum. In Experiment 1, cows were given pulses of saline (control) or 2 micrograms of GnRH infused i.v. during a 0.1-, 1.25-, 5-, 10-, or 20-min period. Concentrations of LH and FSH during 35 min after GnRH infusion were greater than in control cows (P < 0.01), and FSH concentrations were greater when GnRH infusions were for 10 min or less compared with 20 min. In Experiment 2, the effect of GnRH pulse frequency and dose on LH and FSH concentrations, pulse frequency, and pulse amplitude were determined. Exogenous GnRH (0, 2, or 4 micrograms) was infused in 5 min at frequencies of once every hour or once every 4th hr for 3 d. There was a dose of GnRH x frequency x day effect on LH and FSH concentrations (P < 0.01), indicating that gonadotropes are sensitive to changes in pulse frequency, dose, and time of exposure to GnRH. There were more LH pulses when GnRH was infused every hour, compared with an infusion every 4th hr (P < 0.04). Amplitudes of LH pulses were greater with increased GnRH dose (P < 0.05), and there was a frequency x dose x day effect on FSH pulse amplitude (P < 0.0006). We conclude that LH and FSH secretion in the bovine is differentially regulated by frequency and dose of GnRH infusions.     

Effect of metoclopramide on luteinizing hormone secretion in postpartum anestrous cows.

The effect of metoclopramide (MC), a dopamine antagonist on luteinizing hormone (LH), was examined in anestrous primaparous cows. Metoclopramide has been found to be beneficial in overcoming fescue toxicosis; increasing LH secretion stimulates return to ovulatory function after parturition. Consequently, if MC had negative effect on LH secretion, it would indicate that administration of MC to reproducing animals might be limited. Of 14 postpartum (47 to 66 days) cows, 7 were given MC (4 mg/kg of body weight, IV), and 7 served as controls. Blood was obtained via jugular cannulas at 15-minute intervals for 8 hours; MC was given at the end of the first hour, and gonadotropin-releasing hormone (GnRH, 7 mg/kg), was given IV at the end of hour 7 as a challenge stimulus for LH secretion. Prior to GnRH administration, MC did not have significant effect on LH secretion, as judged by mean serum LH concentration, LH pulse frequency, and LH pulse amplitude. Administration of MC resulted in greater (P less than 0.05) LH response to GnRH, indicating enhanced secretory ability when the pituitary gland was challenged. Serum prolactin concentration was increased (P less than 0.01) by MC administration. Therefore, MC did not have adverse effect on LH secretion in postpartum cows.     

Effect of naloxone on serum luteinizing hormone, cortisol and prolactin concentrations in anestrous beef cows

Two experiments were conducted with the opioid antagonist naloxone to determine the effect of opioid receptor blockade on hormone secretion in postpartum beef cows. In Exp. 1, nine anestrous postpartum beef cows were used to measure the effect of naloxone on serum luteinizing hormone (LH), cortisol and prolactin concentrations. Cows received either saline (n = 4) or 200 mg naloxone in saline (n = 5) iv. Blood samples were collected at 15-min intervals for 2 h before and after naloxone administration. Serum LH concentrations increased (P less than .01) in naloxone-treated cows from 1.8 +/- .04 ng/ml before treatment to 3.9 +/- .7 ng/ml and 4.2 +/- .5 ng/ml at 15 and 30 min, respectively, after naloxone administration. In contrast, LH remained unchanged in saline-treated cows (1.6 +/- .3 ng/ml). Serum cortisol and prolactin concentrations were not different between groups. In Exp. 2, 12 anestrous postpartum beef cows were used to examine the influence of days postpartum on the serum LH response to naloxone. Four cows each at 14 +/- 1.2, 28 +/- .3 and 42 +/- 1.5 d postpartum received 200 mg of naloxone in saline iv. Blood samples were taken as in the previous experiment. A second dose of naloxone was administered 2 h after the first, and blood samples were collected for a further 2 h. Serum LH concentrations increased (P less than .01) only in cows at 42 d postpartum.(ABSTRACT TRUNCATED AT 250 WORDS)     

Pituitary receptors for GnRH and estradiol, and pituitary content of gonadotropins in beef cows. II. Changes during the postpartum period

Pregnant beef heifers (n = 24) were assigned randomly to four groups and slaughtered at day 1, 15, 30 or 45 postpartum. The day prior to slaughter blood samples were taken from each cow every 15 min for 8 hr. The anterior pituitary gland, preoptic area (POA) and medial basal hypothalamus (HYP) were collected from each cow. Contents of gonadotropin-releasing hormone (GnRH) in extracts of POA and HYP, and luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in extracts of anterior pituitary were quantified by radioimmunoassay. In the anterior pituitary gland, membrane receptors for GnRH were quantified by a standard curve technique and cytosolic receptors for estradiol were quantified by saturation analysis. Concentrations of LH, FSH and prolactin in serum were quantified by radioimmunoassay. Only one cow of eight had a pulse of LH during the 8 hr bleeding period on day 1 postpartum. This increased to 8 pulses in 6 cows on day 30 postpartum. Contents of GnRH in POA (15.0 +/- 3.2 ng) and HYP (14.0 +/- 2.0 ng) did not change significantly during the postpartum period. Pituitary content of LH was low following parturition (.2 +/- .1 mg/pituitary) and increased significantly through day 30 postpartum (1.2 +/- .1 mg/pituitary). Pituitary content of FSH did not change over the postpartum period. Receptors for both GnRH (.9 +/- .2 pmoles/pituitary) and estradiol (5.0 +/- .9/moles/pituitary) were elevated on day 15 postpartum, possibly increasing the sensitivity of the anterior pituitary gland to these hormones and leading to an increased rate of synthesis of LH that restored pituitary content to normal by day 30 postpartum.     

Patterns of tonic luteinizing hormone release and ovulation frequency in suckled anestrous beef cows following varying intervals of temporary weaning

Thirty postpartum Brahman crossbred cows were utilized to determine the effects of varying intervals of temporary weaning on tonic LH secretion and ovulation. Cows were assigned randomly on day 17–21 postpartum to one of five groups: 1) Suckled Ad libitum, 2) 48-hr weaned, 3) 72-hr weaned, 4) 96-hr weaned, or 5) 144-hr weaned. The mean maximal rise in LH pulse frequency due to weaning occurred within 2 days and averaged 221 percent of time 0 values. The frequency of LH pulses was greater (P<.06) in weaned than in suckled controls. This temporal increase was self-limiting, displaying an acute rise followed by a variable rate of decline in all groups. However, pulse frequency remained elevated relative to suckled controls for the longest period of time for weaning durations of 96 and 144 hr (P<.10). In 48-hr and 72-hr weaned cows, a rapid reversal of the initial increase in LH pulse frequency was observed following calf return. A significant linear regression (y = 1.9 ± .64x; P<.03) described the increase in LH pulse frequency that occurred in cows which ovulated following weaning. Nonovulators were sensitive to calf return and responded by exhibiting a linear decline (y = 2.87 − .43x; P<.04) in LH pulse frequency following this event. The amplitude of LH pulses increased (P<.02) during the period after calf return in ovulators, but did not change in nonovulators. Percentage ovulating by day 10 increased (P<.05) with increased weaning duration past 72 hr. We conclude that calf return before 96 hr markedly attenuates weaning-induced increases in LH secretion and ovulation.     

Endocrine changes in sows exposed to elevated ambient temperature during lactation

Seven sows were placed into one of two environmental chambers at 22 C, 5 d prior to farrowing. On day 9 of lactation, one chamber was changed to 30 C (n = 4) and the other remained at 22 C (n = 3). On days 24 and 25, blood samples were collected every 15 min for 9 hr and 7 hr, respectively. On day 24, thyrotropin releasing hormone (TRH) and gonadotropin releasing hormone (GnRH) were injected iv at hour 8. On day 25 naloxone (NAL) was administered iv at hour 4 followed 2 hr later by iv injection of TRH and GnRH. Milk yield and litter weights were similar but backfat thickness (BF) was greater in 22 C sows (P less than .05) compared to 30 C sows. Luteinizing hormone (LH) pulse frequency was greater (P less than .003) and LH pulse amplitude was less (P less than .03) in 22 C sows. LH concentrations after GnRH were similar on day 24 but on day 25, LH concentrations after GnRH were greater (P less than .05) for 30 C sows. Prolactin (PRL) concentrations were similar on days 24 and 25 for both groups. However, PRL response to TRH was greater (P less than .05) on both days 24 and 25 in 30 C sows. Growth hormone (GH) concentrations, and the GH response to TRH, were greater (P less than .0001) in 30 C sows. Cortisol concentrations, and the response to NAL, were less (P less than .03) in 30 C sows. NAL failed to alter LH secretion but decreased (P less than .05) PRL secretion in both groups of sows. However, GH response to NAL was greater (P less than .05) in 30 C sows. Therefore, sows exposed to elevated ambient temperature during lactation exhibited altered endocrine function.     

Luteinizing hormone secretion in hypophysial stalk-transected gilts given hydrocortisone acetate and pulsatile gonadotropin-releasing hormone.

The site within the hypothalamic-pituitary axis at which cortisol acts to inhibit luteinizing hormone (LH) secretion was investigated in female pigs. Six ovariectomized, hypophysial stalk-transected (HST) gilts were given 1 microgram pulses of gonadotropin releasing-hormone (GnRH) iv every 45 min from day 0 to 12. On days 6-12, each of 3 gilts received either hydrocortisone acetate (HCA; 3.2 mg/kg body weight) or oil vehicle im at 12-hr intervals. Four ovariectomized, pituitary stalk-intact gilts served as controls and received HCA and pulses of 3.5% sodium citrate. Jugular blood was sampled daily and every 15 min for 5 hr on days 5 and 12. Treatment with HCA decreased serum LH concentrations and LH pulse frequency in stalk-intact animals. In contrast, serum LH concentrations, as well as the frequency and amplitude of LH pulses, were unaffected by HCA in HST gilts and were similar to those observed in oil-treated HST gilts. We suggest that chronically elevated concentrations of circulating cortisol inhibit LH secretion in pigs by acting at the level of the hypothalamus.     

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)     

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.     

黄牛孪生处理前后发情周期的外周血中GnRH水平

在母牛产后第1次发情后第10d，给5头发情周期正常的母牛安装颈静脉血管导管；在母牛产后第2个发情周期进行挛生处理，即在产后第2次发情后第10d注射PMSG2000IU，2d后注射氯前列烯醇0．4mg第3次发情后注射抗PMSG抗体2000IU。自第1个发情周期第10d开始，每天间隔10min连续采血2h。发情当天间隔30min采血一次，直到第3次发情结束为止。挛生处理后采血间隔时间和方法与处理前不同。应用酶免疫测定方法测定血清中促性腺激素释放激素水平。结果表明：（1）牛外周血中GnRH释放具有脉冲性；（2)PMSG处理后外周血中GnRH水平升高，表现为脉冲频率增加，脉冲峰值升高。     

Elicitation of release of luteinizing hormone by N-methyl-d,l-aspartic acid during three paradigms of suppressed secretion of luteinizing hormone in the female pig.

Two experiments were conducted to determine the minimal effective dose during lactation and site of action of N-methyl-d,l-aspartic acid (NMA) for elicitation of release of luteinizing hormone (LH) in female pigs. In the first experiment, three doses of NMA were given to lactating primiparous sows in which endogenous LH was suppressed by suckling of litters. In the second experiment, ovariectomized gilts were pretreated with estradiol benzoate or porcine antisera against GnRH to suppress LH and then given NMA to determine if it elicited secretion of LH directly at the anterior pituitary or through release of GnRH. In experiment 1, 3 lactating sows (17 +/- 1.5 d postpartum) were each given three doses of NMA (1.5, 3.0 and 5.0 mg/kg body weight [BW]; IV) on 3 consecutive days in a Latin Square design. Blood samples were collected every 10 min from -1 to 1 hr from injection of NMA. NMA at 1.5 and 3.0 mg/kg did not affect (p greater than .5) secretion of LH; however, 5 mg NMA/kg elicited a 114% increase (p less than .001) in circulating levels of LH during 1 hr after treatment. In experiment 2, 8 ovariectomized gilts were given either estradiol benzoate (EB; 10 micrograms/kg BW; IM n = 4) to suppress release of GnRH or porcine antiserum against GnRH (GnRH-Ab; titer 1:8,000; 1 ml/kg BW; IV; n = 4) to neutralize endogenous GnRH. Gilts infused with GnRH-Ab were given a second dose of antiserum 24 hr after the first. Gilts were then given NMA (10 mg/kg BW; IV) 33 hr after EB or initial GnRH-Ab. Blood samples were drawn every 6 hr from -12 to 24 hr from EB or GnRH-Ab treatments, and every 10 min from -2 to 2 hr from NMA. Serum LH declined (p less than .001) after EB (from 1.87 +/- .2 ng/ml at 12 hr before EB to 0.46 +/- .02 ng/ml during 24 hr after EB) and GnRH-Ab (from 1.97 +/- .1 to 0.59 +/- .02 ng/ml). In gilts treated with EB, the area under the curve (AUC) for the LH response (ng.ml-1.min) 1 hr after NMA (38.7 +/- 3) was significantly greater (p less than .01) than the 1 hr prior to NMA (21.3 +/- 1.5). Treatment with NMA had no effect (p greater than .5) on secretion of LH in gilts infused with GnRH-Ab.(ABSTRACT TRUNCATED AT 400 WORDS)     

Opioid modulation of tonic luteinizing hormone release in ovariectomized dairy cows

A possible role of endogenous opioid peptides (EOP) in regulating the release of luteinizing hormone (LH) in the absence of ovarian influence was investigated. Experiments were conducted on three lactating Holstein-Friesian dairy cows, 20-27 days after ovariectomy. The cows were bled before and after a single intravenous (i.v.) injection of either 250 mg of naloxone (EOP antagonist) or 300 mg of morphine (EOP agonist) or a combination of the two in Experiments 1, 2 and 3, respectively. The mean and basal LH concentrations and the LH pulse frequency and amplitude were compared before and after each treatment in each cow. Naloxone induced an immediate rise in LH concentration by 60-300% above the preceding baseline values. This rise lasted for 15-30 min in each cow, after which the normal rhythmic LH release continued. One cow (A) suffered discomfort and respiratory distress 15-25 min after naloxone administration and the mean and basal LH concentration dropped significantly. Morphine significantly reduced the mean LH concentration by decreasing the number and amplitude of LH pulses and the basal LH values in two cows, although the decrease in one was not significant. The mean LH concentration in each cow remained unaffected by the combined treatment of morphine and naloxone. In conclusion, the elevation of LH concentration by naloxone, the suppression of LH release by morphine and the reversal by morphine and naloxone of each other's effects suggest that EOP could be involved in the control of LH release in cows in the absence of ovarian influence.     

Effects of body condition and estradiol on luteinizing hormone secretion in post-partum beef cows

An experiment was conducted to test the hypothesis that the effect of body fatness on LH pulsatility in post-partum cows is entirely independent of the negative feedback effects of ovarian steroids. Forty beef cows were fed in the last 100 d of gestation so that they achieved either a thin (mean score 1.97) or fat (mean score 2.79) body condition (0 to 5 scale) at calving and were fed after calving to maintain live weight and body condition. At 15 (sd 3.7) d post partum all cows were ovariectomised and half from each body condition score treatment group received a subcutaneous estradiol implant (+EST) while the remainder received no implant (−EST). At weeks 5 and 9 post-partum blood samples were collected via jugular catheter every 20 minutes for 10 hr on two consecutive d and on the third d cows were injected via the jugular vein with 2.5 μg GnRH. Blood samples were collected every 15 minutes for 1 hr before and 2 hr after GnRH injection. At 5 and 9 weeks the fatter cows had significantly higher mean LH concentrations, baseline LH concentrations, LH pulse amplitudes and pulse frequencies (P<0.01). Implantation with estradiol in both fat and thin cows reduced mean LH concentrations, baseline LH concentrations, LH pulse amplitudes and pulse frequencies (P<0.001). The lack of interaction between body condition and the presence or absence of estradiol implies that the effect of body condition on LH release is independent of ovarian steroid feedback mechanisms. Fat cows showed a greater release of LH in response to exogenous GnRH (P<0.01) than thin cows while implantation with estradiol in both fat and thin cows decreased (P<0.01) LH release. The pituitary responsiveness to GnRH with the −EST cows was greater at 9 compared to 5 weeks, but there was no difference with time in the +EST cows. However, there was no such interaction in endogenous LH pulse amplitude suggesting that in the absence of estradiol the magnitude of GnRH pulses declined with time post-partum.     

N-methyl-d,l-aspartate modulation of pituitary hormone secretion in the pig: Role of opioid peptides

Sixteen ovariectomized (OVX) mature gilts, averaging 139.6 ± 3.1 kg body weight (BW) were assigned randomly to receive either progesterone (P, 0.85 mg/kg BW, n=8) or corn oil vehicle (OIL, n=8) injections im twice daily for 10 d. On the day of experiment, all gilts received either the EAA agonist, N-methyl-d,l-aspartate (NMA; 10 mg/kg BW, iv) alone or NMA plus the EOP antagonist, naloxone (NAL, 1 mg/kg BW, iv), resulting in the following groups of 4 gilts each: OIL-NMA, OIL-NMA-NAL, P-NMA and P-NMA-NAL. Blood samples were collected via jugular cannula every 15 min for 6 hr. All pigs received NMA 5 min following pretreatment with either 0.9% saline or NAL 2 hr after blood collection began and a GnRH challenge 3 hr after NMA. Administration of NMA suppressed (P<0.03) LH secretion in OIL-NMA gilts and treatment with NAL failed to reverse the suppressive effect of NMA on LH secretion in OIL-NMA-NAL gilts. Similar to OIL-NMA gilts, NMA decreased (P<0.03) mean serum LH concentrations in P-NMA gilts. However, in P-NMA-NAL gilts, serum LH concentrations were not changed following treatment. All gilts responded to GnRH with increased (P<0.01) LH secretion. Additionally, administration of NMA increased (P<0.01) growth hormone (GH) and prolactin (PRL) secretion in both OIL-NMA and P-NMA gilts, but this increase in GH and PRL secretion was attenuated (P<0.01) by pretreatment with NAL in OIL-NMA-NAL and P-NMA-NAL gilts. Serum cortisol concentrations increased (P<0.01) in all gilts and the magnitude of the cortisol response was not different among groups. In summary, results of the present study confirmed previous findings that NMA suppresses LH secretion in both oil- and P-treated OVX gilts, but we failed to provide definitive evidence that EOP are involved in the NMA-induced suppression of LH secretion. However, NMA may, in part, activate the EOP system which in turn increased GH and PRL secretion in the gilt.     

Adenohypophyseal receptors for LHRH, pituitary content of gonadotropins and plasma concentrations of LH in cyclic, pregnant and postpartum beef cows

Adenohypophyseal concentrations of LHRH receptors, pituitary content of LH and FSH, and plasma concentrations of LH were determined in thirty Hereford, Angus or Hereford-Angus heifers that were randomly assigned by breed and weight to five periods including day 3 of the estrous cycle (CY), pregnant day 120 (P120), 200 (P200), 275 (P275), or day 2 postpartum (PP). Jugular blood samples were collected at 10-min intervals for 8 hr from all cows. Within 2 hr after completion of blood sampling, animals were slaughtered and the pituitary gland frozen at −196 C. LH pulse frequency/8 hr was reduced (P<.05) during gestation (.5, .2, and 1.5 ± .5/8 hr, for P120, P200, and P275, respectively) and PP (.5 ± .5/8 hr) compared to CY (7.8 ± .5/8 hr). Frequency of LH pulses/8 hr was not different (P>.1) among P120, P200 or PP periods but was different (P<.05) between P200 and P275. There were no differences in LH pulse height (P>.1) among periods; however, pulse amplitude was greatest (P<.05) at P120 (1.3 ± .2 ng/ml) and lowest between P200 and PP (.6 to .8 ± .2 ng/ml). Baseline concentrations of plasma LH did not differ (P>.1) among P and PP periods (.3 ± .1 ng/ml), but were lower (P<.05) than in CY animals (.7 ± .1 ng/ml). Concentration of adenohypophyseal LHRH receptors was approximately two-fold greater (P<.05) at P120 (25.85 ± 2.2 fmol/mg) than at all other periods (9.5 to 14.9 ± 2.2 fmol/mg). Pituitary content of LH was greatest at P120 (1.56 ± .11 ug/mg) and lowest (P<.05) at P275 and PP (0.46 to 0.52 ± .11 ug/mg). Pituitary content of FSH was greatest (P<.05) in P (12.7 to 17.0 ± 1.4 ug/mg) and PP (18.3 ± 1.4 ug/mg) vs CY (5.0 ± 1.4 ug/mg) cows and increased from P120 to PP (P<.05). Results indicate that physiological changes occurring during gestation may have an effect on subsequent function of the adenohypophysis in beef cows.     

A study on ghrelin and LH secretion after short fasting and on ghrelin levels at perioestrual period in dairy cattle

In two experiments, we studied (a) the changes of LH secretion in heifers under different feeding schedules and (b) total ghrelin concentration at oestrus in cows and heifers. In experiment one, synchronized heifers were allocated in three groups (R, regularly fed controls; F, fasted; and F‐F fasted‐fed). One day after the completion of the oestrous induction protocol, group F and F‐F animals stayed without feed for 24 hr; thereafter, feed was provided to R and F‐F cattle; 2 hr later, GnRH was administered to all animals. Blood samples were collected for ghrelin, progesterone, LH and cortisol concentrations. Fasting caused increased ghrelin concentrations in groups F and F‐F, while in response to GnRH, LH surge was significantly attenuated in groups F and F‐F compared to R. In experiment 2, lactating cows and heifers were used. On day 9 of a synchronized cycle, PGF2α was administered, and blood samples were collected twice daily until the third day after oestrus and analysed for progesterone, estradiol, ghrelin, glucose and BHBA concentrations. No difference was recorded between groups in steroids and BHBA concentrations. In comparison to mid‐luteal values, ghrelin concentrations significantly increased at perioestrual period in cows, but not in heifers. This study provides evidence that starving‐induced elevated ghrelin concentrations can have suppressing effect on LH secretion, even after ghrelin's restoration to basal values and that during oestrus, ghrelin secretion is differently regulated in cows and heifers, likely being independent from oestradiol concentrations. Further research is required to identify the determining factors that drive the different regulation of ghrelin secretion in cows and heifers.