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)     

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.     

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.     

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)     

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.     

Reduced postpartum anestrus of suckled beef cows treated with microencapsulated luteinizing hormone-releasing hormone analog.

This study evaluated the effect of microencapsulated LHRH agonist (D-Trp6-LHRH) on gonadotropin release and occurrence of estrus in early postpartum beef cows. Angus cows (n = 54) were assigned randomly to two treatment groups at d 5 postpartum. Group 1 received a single i.m. injection of D-Trp6-LHRH (LHRH-A) encapsulated in poly-DL-lactide-coglycolide, calculated to release 15 micrograms of LHRH-A per day for 30 d (n = 23). Group 2 received vehicle only (control, n = 31). Blood samples (15-min intervals for 6 h) were obtained on d 5, 10, 20, 30, and 40 postpartum for evaluation of LH and FSH concentrations (n = 12 per group). Days to first postpartum estrus were reduced by treatment with LHRH-A (Group 1, 43.7 +/- 4.2 d vs Group 2, 55.9 +/- 4.7 d; P < .05). However, days to conception were similar between groups (68.9 +/- 7.9 vs 76.7 +/- 6.7 d, respectively). On the day of treatment, cows treated with LHRH-A had higher mean concentrations of LH and FSH than did controls (8.3 +/- 1.4 vs 2.0 +/- .4 ng/mL for LH and 211.0 +/- 8.6 vs 51.2 +/- 2.7 ng/mL for FSH (P < .05). There were no differences in mean concentrations of LH or FSH between treatment groups on d 10, 20, 30, and 40 postpartum. Cows given LHRH-A had more (P < .05) LH pulses on d 10 and 30 postpartum than did controls. This study demonstrated that microencapsulated D-Trp6-LHRH reduced the postpartum anestrous interval in suckled beef cows.     

Relationship between weaning and secretion of luteinizing hormone, cortisol and transcortin in beef cows

The effects of suckling on secretion of luteinizing hormone, cortisol and transcortin were investigated in anovulatory postpartum cows. On d 35 postpartum, calves were separated from 12 cows to prevent suckling and eight calves continued to suckle their dams ad libitum. Between 35 and 41 d postpartum, samples of jugular blood were collected every 15 min for two periods of 6 h/d. In non-suckled cows, frequency of pulses and basal luteinizing hormone increased but amplitude of pulses did not change. Concentrations of total cortisol in serum of cows were not altered during 3 d after weaning calves and did not differ among intervals before, during and after a suckling event. Affinity of transcortin for cortisol was not affected by postpartum interval or treatment. Capacity of transcortin to bind cortisol tended to increase after weaning. We found no evidence to support the hypothesis that suckling reduces binding capacity of transcortin or increases unbound cortisol. Differences in preovulatory secretion of luteinizing hormone between suckled and non-suckled cows could not be accounted for by differences in secretion of cortisol. In beef cows that are fed to satisfy requirements for energy and have average body condition, we conclude that negative modulation of luteinizing hormone by suckling is not mediated by cortisol.     

Relationships among concentrations of four opioid neuropeptides and luteinizing hormone-releasing hormone in neural tissues of beef cows following early weaning

Acute changes associated with removal of the inhibition of estrus caused by suckling were examined in beef cows. Calves were weaned during the fifth week after parturition and cows were slaughtered at 0 (n = 8), 36 (n = 8) or 72 h (n = 8) after calf removal. Tissues of preoptic area (POA), hypothalamus (HYP), pituitary stalk-median eminence (SME) and pituitary neurointermediate lobe (NIL) were obtained for analyses of luteinizing hormone-releasing hormone (LHRH) and four opioid neuropeptides. In addition, one-half of each SME was superfused in vitro for measurement of basal and potassium-induced release of LHRH. The following opioid neuropeptides were quantified: methionine-enkephalin (Met-Enk), beta-endorphin (beta-EP), dynorphin-A, 1-17 (DYN-17) and dynorphin-A, 1-8 (DYN-8). All four opioid neuropeptides were most concentrated in the pituitary NIL. Luteinizing hormone-releasing hormone was most concentrated in the SME tissue, which also contained substantial concentrations of Met-Enk and beta-EP, but very little DYN-17 or DYN-8. In addition, weaning increased the weight of NIL between 0 and 36 h (P less than .05), and the concentrations of LHRH, Met-Enk, and DYN-17 in the combined POA + HYP (P less than .05) tissue between 36 and 72 h. No differences occurred among groups in SME content of LHRH or in vitro release of LHRH from the superfused SME. Although they were not affected by weaning, within-cow correlations among parameters revealed that: 1) concentrations of DYN-17 and DYN-8 were always positively correlated (P less than .05); 2) concentrations of LHRH were positively correlated with Met-Enk (P less than .01), beta-EP (P less than .05) and DYN-17 (P less than .05) in the combined POA + HYP tissue; 3) LHRH concentrations in SME tissue were negatively related to POA + HYP concentrations of Met-Enk (P less than .01) and beta-EP (P less than .05), but not of LHRH or DYN-17 and 4) in vitro release of LHRH from the pituitary SME was correlated with concentrations of DYN-8 in various tissues including the SME (P less than .01). In summary, bovine neural tissues differ widely in concentrations of the four opioid neuropeptides with NIL tissue having the greatest concentrations. Weaning calves at 36 and 72 h before slaughter caused parallel changes in LHRH, Met-Enk and DYN-17 in preoptic and hypothalamic tissues.(ABSTRACT TRUNCATED AT 400 WORDS)     

Comparison of the effectiveness of ovulation synchronization protocol in anestrous and cycling beef cows

Applicability of ovulation synchronization protocol using GnRH and PGF(2alpha) (PGF) injection to anestrous beef cows remains controversial. We compared the effectiveness of the protocol in the anestrous stage of the beef cow with that in the cycling stage using the same animals. Ovaries of five Japanese Black and three Japanese Shorthorn cows were ultrasonographically examined, and blood samples were collected daily for hormonal analyses. Each animal received the protocol twice (Day -6 to -8: GnRH, Day 0: PGF, Day 2: GnRH). Additional blood samples were taken before and after GnRH injection for LH and FSH measurements to evaluate the pituitary function. For the ovarian status at the onset of the protocol cows were divided into anestrous (n=8) and cycling (n=8) stages. There was no significant difference in size of the dominant follicle at the first and second GnRH injections, and in the magnitude of the pituitary response to GnRH between the two stages. However, the size of the corpus luteum and progesterone concentrations at the PGF injection in the anestrous stage were significantly smaller and lower (P<0.01), respectively, and ovulation synchronization rate in the anestrous stage was significantly lower (P<0.05) than in the cycling stage. In conclusion, ovulation synchronization protocol in anestrous beef cows has limited effectiveness.     

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.     

Gonadotropin-releasing hormone-induced ovulation and luteinizing hormone release in beef heifers: effect of day of the cycle

The COSynch protocol has been used to synchronize ovulation and facilitate fixed-time AI in beef cattle. Establishment and maintenance of pregnancy was negatively affected, in previous studies, by GnRH-induced ovulation of small dominant follicles (/=10 mm) and increased ovulatory response after GnRH 2.     

Cortisol and luteinizing hormone after adrenocorticotropic hormone administration to postpartum beef cows

Cortisol and luteinizing hormone (LH) were measured in serum after the administration of adrenocorticotropic hormone (ACTH) to suckled (S) and nonsuckled (NS) beef cows. Blood was sampled on 2 consecutive days every 2 weeks for four bleeding periods starting 14 days after calving. Cows were injected with 200 IU ACTH or saline in a 2-day switchback design. Serum was collected before ACTH or saline injection and at 30-min intervals thereafter for 8 hours. Average cortisol concentrations in serum were similar in S and NS cows (6.4 +/- .6 and 6.1 +/- .8 ng/ml, respectively) after saline. Average cortisol concentrations in serum collected during an 8-hr period after ACTH on days 14, 28, 42 and 56 postpartum were 24.7 +/- 2.4, 31.8 +/- 3.5, 36.4 +/- 4.2 and 40.7 +/- .5 ng/ml, respectively, for S cows, and 31.1 +/- 2.9, 44.7 +/- 5.2, 45.0 +/- 5.7 and 46.0 +/- 5.4 ng/ml, respectively, for NS cows. Cortisol response to ACTH, measured as area under the response curve, was greater (P less than .05) in NS than in S cows. Amount of cortisol released by 200 IU ACTH was maximal by days 28 to 29 postpartum in NS cows, but the response increased gradually between days 14 to 15 and days 56 to 57 in S cows. overall, LH in serum averaged .55 +/- .08 ng/ml for S cows and .92 +/- .06 ng/ml for NS cows after saline, and .49 +/- .07 ng/ml for S cows and .94 +/- .06 ng/ml for NS cows after ACth. Although mean and peak serum LH concentrations did not differ between cows given ACTH and those given saline, the number of LH peaks and the number of cows having LH after saline. Mean serum LH concentrations were lower (P less than. 05) in S than in NS cows at 28 days postpartum. The number of LH peaks was lower (P less than .05) and the magnitude of the largest LH peak tended to be lower (P less than .06) in S cows at all sampling periods.     

Synchronized estrus and fertility of beef cows after weaning calves for short intervals

In Exp. 1, the objective was to determine if interval of separating calves from cows (24 or 48 h) immediately before insemination affects detection and precision of estrus and pregnancy rates of lactating beef cows implanted with norgestomet. Separation of calves from cows for 24 h (n = 418) lengthened intervals to estrus, did not affect precision of estrus, reduced success of detecting estrus and lowered pregnancy rates relative to positive controls (48 h separation, n = 508). Cows with poor body condition, and not suckled for 24 h, conceived at lower rates than cows with similar condition that were not suckled for 48 h. Adverse effects of separation for only 24 h on fertility are apparently due to inadequate intervals between estrus and insemination at 48 h after removing implants. In Exp. 2, the objective was to determine effects of separating calves from cows for 48 h immediately before insemination on detection and precision of estrus and on pregnancy rate of ovulatory lactating beef cows injected twice with prostaglandin F2 alpha (PGF2 alpha). Weaning increased detection of estrus but overall pregnancy rates did not differ between suckled (n = 256) and nonsuckled (n = 221) cows. But, weaning calves improved pregnancy rates of young (2 to 3 yr) cows and reduced fertility among middle (4 to 6 yr)-aged cows. Increased pregnancy rates after weaning calves for 48 h are due largely to greater detection of estrus and inseminating more cows.(ABSTRACT TRUNCATED AT 250 WORDS)     

Pituitary concentrations of gonadotropins and receptors for GnRH in suckled beef cows at various intervals after calving

Mature beef cows were slaughtered at 5 (n = 6), 10 (n = 6), 20 (n = 6) or 30 (n = 5) d after calving to identify endocrine events that may affect the duration of postpartum anestrus. Additional cows (n = 6) were slaughtered 12 to 14 d after their first postpartum estrus (luteal phase cows). Anterior pituitary concentrations of luteinizing hormone (LH) were low at d 5 (383 +/- 69 micrograms/g), averaged 445 +/- 103 and 682 +/- 207 micrograms/g at d 10 and 20, respectively, and were elevated (P less than .05) by d 30 (1,097 +/- 174 micrograms) to a concentration similar to luteal phase cows (1,208 +/- 148 micrograms/g). Concentrations of follicle-stimulating hormone (FSH) averaged 12.4 +/- 1.1, 9.6 +/- 2, 8.6 +/- 1.8 and 7.4 +/- 3.3 mg/g at d 5, 10, 20 and 30, respectively. Affinity (1.6 +/- .2 X 10(9) M-1) of anterior pituitary receptors for the GnRH (gonadotropin-releasing hormone) analog (DAla6; des-Gly10, [D-Ala6]-LH-RH ethylamide) and weights (2.1 +/- .1 g) of the anterior pituitaries did not differ among groups (P greater than .05). Number of receptors for GnRH averaged 37 +/- 7, 39 +/- 9, 25 +/- 5 and 23 +/- 5 X 10(-14) M/mg protein at d 5, 10, 20 and 30, respectively. Anterior pituitaries from luteal phase cows contained 22 +/- 2 X 10(-14) M/mg protein of receptors for GnRH.(ABSTRACT TRUNCATED AT 250 WORDS)     

Synchronization of ovulation using GnRH or hCG with the CO-Synch protocol in suckled beef cows.

The objectives of this study were to evaluate replacing GnRH with hCG and the effects of 48-h calf removal (CR) on pregnancy rates of cows synchronized with the CO-Synch protocol. Suckled beef cows (n = 467) at two locations were assigned to treatment by breed, age, and calving date. Treatment included either GnRH with (n = 121) or without CR (n = 117) or hCG with (n = 115) or without CR (n = 114) using the CO-Synch protocol. On d 0 and 9, cows received either hCG (2,500 IU, i.m.) or GnRH (100 microg, i.m.), and on d 7 all cows received PGF2alpha (25 mg). At one location, blood samples were collected from all cows (n = 203) on d -14, -7, 0, 7, 9, and 16. Calves were removed on d 7 and returned on d 9 (48 h) from approximately half of the cows that received GnRH or hCG. Cows that were detected in estrus between d 6 and 9 were bred approximately 12 h later and received no further injections. Cows not observed in estrus by d 9 received a second injection of either GnRH or hCG and were timed-inseminated. The AI pregnancy rates for GnRH-treated cows with or without CR and hCG-treated cows with or without CR were 46, 49, 35, and 34%, respectively (P = 0.44). Pregnancy rates of cows differed by treatment x age interaction (P = 0.07), hormone (P = 0.09), and hormone x age (P = 0.01) but not by CR (P = 0.66) or CR x age (P = 0.33). Among 2-yr-olds, pregnancy rates were higher for cows treated with hCG without CR than for cows that received GnRH with calf removal, whereas cows treated with hCG with CR and GnRH without CR were intermediate. In addition hCG-treated 2-yr-olds had higher pregnancy rates than GnRH-treated 2-yr-olds regardless of calf presence, but the reverse was true for older cows. Overall, GnRH-treated cows (48%) had a higher (P = 0.09) pregnancy rate than hCG-treated cows (34%). Among anestrous cows, GnRH and hCG were similar (P = 0.40) in their ability to induce ovulation and corpus luteum formation after the first and second injections of GnRH (31 and 76%, respectively) or hCG (39 and 61%, respectively). More (P = 0.001) hCG-treated cows exhibited short estrous cycles following timed AI. We conclude that hCG is not a suitable replacement for GnRH to synchronize ovulation with the CO-Synch protocol in multiparous cows, although further evaluation among primiparous cows is warranted using hCG with the CO-Synch protocol.