Luteinizing hormone in nutrient-restricted ovariectomized ewes

This study was designed to evaluate profiles of serum concentrations of LH, pituitary content of LH and GnRH receptors, and hypothalamic content of GnRH in undernourished, ovariectomized ewes. In earlier studies, pulsatile secretion of LH diminished as duration of undernutrition progressed in prepubertal or adult ovariectomized ewes. Ewes having similar body condition scores (CS) of 5 to 9 (1 = extremely thin, 5 = moderate, 9 = obese) were fed maintenance or low-energy diets (100% and 60% of NRC requirements, respectively). Blood samples for analysis of LH were collected at 15-min intervals for 4 h at initiation of the project and immediately prior to slaughter. Serum concentrations of LH did not differ (P greater than .05) among groups at the initial sampling period. At slaughter, ewes with CS less than or equal to 2 (n = 7) had lost 26.8 kg (42% of initial weight). Ewes with CS greater than or equal to 3 (n = 12) had lost an average of 13.7 kg (18% of initial weight). Concentrations of LH in ewes with final CS greater than or equal to 3 was similar (P greater than .05) to that observed during the initial sampling period. However, release of LH was reduced (P less than .01) in ewes with CS less than or equal to 2 compared with ewes with CS greater than or equal to 3 (2.6 vs 9.5 and 3.2 vs 10.5 ng/ml for basal and mean concentrations, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of enclomiphene on estradiol-induced changes in LH secretion in ewes

Experiments were conducted to characterize the ability of the antiestrogen enclomiphene (ENC) to block the effects of estradiol on secretion of LH in ovariectomized ewes. To determine whether ENC could block an estradiol-induced LH surge, ewes (n = 4/group) were administered 10 to 250 mg ENC followed 30 min later by 25 micrograms estradiol. Ten or 25 mg ENC suppressed the estradiol-induced LH surge in one of four ewes, whereas 100- or 250-mg doses suppressed the LH surge in three and four of four ewes, respectively. In ewes that received a single treatment of 100 mg ENC plus 25 micrograms estradiol, serum concentrations of LH remained below 1 ng/ml for 3 wk. Compared with untreated ewes, the number of pituitary GnRH receptors was elevated (P less than .05) at 12 d and 28 d, but pituitary content of LH had decreased (P less than .05) by 28 d in ewes treated with 100 mg ENC. To determine whether ENC could block the inhibitory effects of estradiol on serum concentrations of LH, ewes received injections of .03, .1, 1 or 10 mg ENC every 4 d. Half the ewes treated with each dose also received estradiol implants. Injection of .03, .1 or 1 mg ENC alone did not affect serum concentrations of LH, whereas the 10-mg dose decreased serum concentrations of LH below 1 ng/ml by wk 1 of treatment. No dose prevented the inhibition of serum concentrations of LH caused by estradiol implants. In ovariectomized ewes, ENC was antagonistic to estradiol; it prevented the positive effects of estradiol required to induce an LH surge.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of estradiol-17 beta, naloxone and gonadotropin releasing hormone on postpartum secretion of luteinizing hormone in fall-lambing ewes

The interaction among exogenous estradiol-17 beta, naloxone and gonadotropin releasing hormone (GnRH) in the control of luteinizing hormone (LH) secretion was studied in intact postpartum ewes nursing their offspring. One-half of 30 fall-lambing ewes were implanted subcutaneously with an estradiol-17 beta containing Silastic capsule between postpartum d 1 and 12 which doubled their serum concentrations of estradiol (16.0 +/- .1 vs 8.4 +/- .1 pg/ml). Blood samples were collected from implanted and non-implanted ewes at 15-min intervals for 5 h on d 3, 8, 13, 20 and 28 postpartum. Pre-injection samples were collected for 1 h, and ewes were injected with saline, naloxone (NAL;1 mg/kg) or GnRH (100 micrograms/ewe). When averaged across all days and implant groups, serum LH in the three post-NAL samples was higher (P less than .05) than in the three pre-NAL samples (3.6 +/- 1.2 vs .6 +/- .2 ng/ml). Post-GnRH concentrations of serum LH were lower (P less than .05) in estradiol-implanted ewes than in non-implanted ewes on d 8 and 13, but there were no differences in any LH characteristics on d 20 and 28 after implant removal on d 12. In non-implanted ewes, serum LH responses to GnRH increased (P less than .05) eightfold from d 3 (3.8 +/- 1.4 ng/ml) to d 8 (31.6 +/- 1.4 ng/ml), remained elevated through d 20, but declined by d 28 (10.8 +/- 1.4 ng/ml).(ABSTRACT TRUNCATED AT 250 WORDS)     

Basal and GnRH induced secretion of FSH and LH in ovariectomized ewes treated with bovine follicular fluid

Ovariectomized (OVX) ewes were injected with 5 ml of either bovine serum, charcoalextracted bovine follicular fluid (FF), or whole bovine FF. Five hours after this pretreatment, ewes on each pretreatment were injected with either 0, 1, or 5 μg of GnRH. Ewes that were pretreated with either type of FF had decreased concentrations of FSH regardless of dose of GnRH when compared to ewes pretreated with bovine serum. There was no effect of charcoal extraction. There were no differences among the pretreatment groups in LH response to GnRH. In a second experiment, OVX ewes were pretreated (4 ml) with either bovine serum or bovine FF 5 hr prior to GnRH or with bovine FF 42, 30 and 18 hr prior to GnRH. Ewes were injected with either 0 or 5 μg of GnRH. Pretreatment with FF for 5 or 42 hr prior to GnRH resulted in significantly decreased concentrations of FSH both at the time of GnRH treatment and during the following 2 hr. Concentrations of LH did not differ among pretreatment groups. In a third experiment, OVX ewes were pretreated with either bovine serum or bovine FF 30, 18 and 5 hr prior to GnRH. Ewes were injected with either 0, 5 or 50 μg of GnRH. Pretreatment with FF resulted in decreased concentrations of FSH both at the time of GnRH treatment and during the following 2 hr. Concentrations of LH were also decreased at the time of GnRH treatment.     

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.     

Effects of gonadotropin-releasing hormone infused in a pulsatile or continuous fashion on serum gonadotropin concentrations and ovulation in the mare.

Studies were conducted to compare continuous vs pulsatile i.v. infusion of GnRH on serum gonadotropin concentrations and ovulation in seasonally anestrous mares and in cycling mares. Anestrous mares (Exp. 1) received no treatment (control; n = 3), 2, or 20 micrograms of GnRH/h continuous infusion (CI) (n = 4 and n = 6, respectively), or 20 micrograms of GnRH/h pulsatile infusion (PI) (n = 5). After initiation of GnRH infusion, serum LH levels increased earlier, and to a greater extent, in the PI group than in other groups (P less than .05). In contrast, serum FSH concentrations did not differ among groups. The number of days to development of the first 35-mm follicle was not different among GnRH treatment groups; however, mares receiving PI ovulated on d 9.4 of treatment, 2.8 d earlier than those receiving 20 micrograms of GnRH/h CI (P less than .05). Mares given 2 micrograms of GnRH/h CI failed to ovulate spontaneously after 16 d of treatment, but each one ovulated within 2 to 4 d after injection of 2,000 IU of hCG on d 16. Control mares did not ovulate or show any significant follicular development throughout the experiment. Cycling mares (Exp. 2) received no treatment (control; n = 6), 20 micrograms of GnRH/h CI, or 20 micrograms of GnRH/h PI (n = 4) beginning on d 16 of an estrous cycle (d 0 = day of ovulation). Serum LH concentrations in all groups increased after initiation of treatment; however, on the day of ovulation LH concentrations were lower in the CI group than in the PI or control groups (P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of dopamine, norepinephrine and serotonin on secretion of luteinizing hormone, follicle-stimulating hormone and prolactin in ovariectomized, pituitary stalk-transected ewes

Two experiments were conducted in ovariectomized, pituitary stalk-transected ewes to determine if dopamine (DA), norepinephrine (NE) or serotonin (5-HT) alter secretion of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and prolactin (PRL). In experiment 1, ewes were infused (iv) with saline (control), DA (66 micrograms/kg/min), NE (6.6 micrograms/kg/min) or 5-HT (6.6 micrograms/kg/min). Treatments did not alter pulse frequency, but 5-HT increased (P less than .05) amplitude of pulses of LH and mean concentrations of LH, DA and NE were without effect on basal secretion of LH. DA but not NE or 5-HT decreased (P less than .05) the release of LH in response to gonadotropin hormone-releasing hormone (GnRH, 25 micrograms, im). Concentrations of FSH were not affected by treatments. Secretion of PRL was reduced (P less than .05) by treatment with DA and NE but not 5-HT. Each amine reduced (P less than .05) the release of PRL in response to thyrotropin-releasing hormone (TRH; 3 micrograms, im). In experiment 2, ewes were given DA at doses of 0, 0.66, 6.6 or 66.0 micrograms/kg/min, iv. No dose altered basal LH, but each dose reduced (P less than .05) basal and TRH-induced release of PRL. Key findings from these studies include direct pituitary action for: (1) 5-HT enhanced basal secretion of LH, (2) suppression of GnRH-induced secretion of LH by DA. (3) DA and NE inhibition of PRL secretion, and (4) DA, NE and 5-HT inhibition of release of PRL in response to TRH.     

N-methyl-d,l-aspartate stimulates growth hormone and prolactin but inhibits luteinizing hormone secretion in the pig.

The effects of n-methyl-d,l-aspartate (NMA), a neuroexcitatory amino acid agonist, on luteinizing hormone (LH), prolactin (PRL) and growth hormone (GH) secretion in gilts treated with ovarian steroids was studied. Mature gilts which had displayed one or more estrous cycles of 18 to 22 d were ovariectomized and assigned to one of three treatments administered i.m.: corn oil vehicle (V; n = 6); 10 micrograms estradiol-17 b/kg BW given 33 hr before NMA (E; n = 6); .85 mg progesterone/kg BW given twice daily for 6 d prior to NMA (P4; n = 6). Blood was collected via jugular cannulae every 15 min for 6 hr. Pigs received 10 mg NMA/kg BW i.v. 2 hr after blood collection began and a combined synthetic [Ala15]-h GH releasing factor (1-29)-NH2 (GRF; 1 micrograms/kg BW) and gonadotropin releasing hormone (GnRH; .2 micrograms/kg BW) challenge given i.v. 3 hr after NMA. NMA did not alter LH secretion in E gilts. However, NMA decreased (P < .02) serum LH concentrations in V and P4 gilts. Serum LH concentrations increased (P < .01) after GnRH in all gilts. NMA did not alter PRL secretion in P4 pigs, but increased (P < .01) serum PRL concentrations in V and E animals. Treatment with NMA increased (P < .01) GH secretion in all animals while the GRF challenge increased (P < .01) serum GH concentrations in all animals except in V treated pigs. NMA increased (P < .05) cortisol secretion in all treatment groups. These results indicate that NMA inhibits LH secretion and is a secretagogue of PRL, GH and cortisol secretion with ovarian steroids modulating the LH and PRL response to NMA.     

Effect of copper status, supplementation, and source on pituitary responsiveness to exogenous gonadotropin-releasing hormone in ovariectomized beef cows

The effect of Cu status, supplementation, and source on pituitary responsiveness to exogenous GnRH was evaluated using nine multiparous, nonpregnant, nonsuckling, ovariectomized Angus cows (7.1 +/- 3.3 yr; 622.9 +/- 49.8 kg; BCS = 6.0 +/- 0.5). Cows were considered Cu-deficient based on liver Cu concentrations (< 30 mg of Cu/kg of DM) after receiving a low-Cu, forage-based diet supplemented (DM basis) with 5 mg of Mo/kg and 0.3% S for 216 d. Copper-deficient cows were stratified based on age, BW, BCS, and liver Cu concentration and assigned randomly to repletion-phase treatments. Treatments included 1) control (no supplemental Cu); 2) organic (ORG; 100% organic Cu); and 3) inorganic (ING; 100% inorganic CuSO4). Treatments were formulated to meet all NRC recommendations, except for Cu, which was supplemented to ORG and ING cows at 10 mg of Cu/kg of dietary DM. During the 159-d repletion phase, Cu status was monitored via liver biopsy samples, and all cows received exogenous progesterone. A controlled intravaginal drug-release device (replaced every 14 d) was used to maintain luteal phase progesterone as a means to provide negative feedback on the hypothalamic-pituitary axis. During the repletion phase, liver Cu concentrations did not differ between ORG and ING cows at any time. By d 77 of the repletion phase, all supplemented cows were considered adequate in Cu, and liver Cu concentrations were greater in supplemented than in nonsupplemented control cows on d 77 (P < 0.05) and throughout (P < 0.01) the repletion phase. Beginning on d 99, exogenous GnRH was administered to all cows at low (0, 3, and 9 microg; Exp. 1) and high doses (0, 27, and 81 microg; Exp. 2) at six different times. Cows were catheterized every fifth day, and blood samples were collected every 15 min for 1 h before and 4 h after GnRH administration and analyzed for LH concentration. In Exp. 1, Cu status and supplementation did not affect basal or peak LH concentrations, but total LH released tended (P < 0.07) to be greater in Cu-supplemented vs. control cows when 3 microg of GnRH was administered. In Exp. 2, there was no effect of Cu supplementation or source on basal, peak, or total LH released, regardless of GnRH dose. Pituitary LH concentrations did not differ across treatments. In conclusion, Cu status, supplementation, and source did not affect GnRH-induced LH secretion or pituitary LH stores in ovariectomized, progesterone-supplemented cows in this experiment.     

Effects of sodium pentobarbital on release of gonadotropins in ewes immediately after ovariectomy and after treatment with gonadotropin-releas ing hormone

Two experiments were conducted with ewes 9 to 11 days after estrus to determine whether the secretion of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) are controlled differentially. In experiment 1, gonadotropin-releasing hormone (GnRH) was injected (100 (μg/ewe) at time = 0 min into ewes in four treatment groups. The treatment groups (9 ewes/group) were: 1) periodic iv sodium pentobarbital (NaPen) vehicle from 0 min; 2) periodic iv NaPen from 0 min; 3) vehicle iv for 120 min then iv NaPen from 120 min; 4) vehicle iv for 150 min then iv NaPen from 150 min. A surgical plane of anesthesia was maintained from the initiation of NaPen injection until the experiment ended. Jugular blood was sampled at 30-min intervals from ?30 to + 210 min for LH and FSH assays, and profiles of hormone concentrations were compared by time-trend analyses. GnRH released LH (P<.001) and FSH (P<.001), but NaPen did not affect the profiles of hormone concentrations; this indicated that NaPen did not reduce the ability of the pituitary to secrete gonadotropins in response to GnRH. Experiment 2 was a 2x2 factorial with ovariectomy (time = 0 hr) and NaPen as the main effects. One group of ovariectomized (n = 6) and one group of sham ovariectomized (n = 6) ewes were anesthetized only during surgery, while a group of ovariectomized (n = 7) and a group of sham ovariectomized (n = 6) ewes were kept at a surgical plane of anesthesia until 10 hr after surgery. Patterns of LH and FSH were compared in jugular blood collected hourly from 0 hr until 10 hr after surgery and in samples collected at 24 hr intervals from -24 to +72 hr of surgery. After ovariectomy, LH increased (P<.001) hourly and daily, but anesthesia suppressed (hourly, <.001 and daily, P<.005) these increases, which resulted in an interaction (hourly, P<.001 and daily, P<.01) of ovariectomy and anesthesia. FSH after ovariectomy increased hourly and daily (hourly, P<.02 and daily, P<.001), but the effect of anesthesia and interaction of ovariectomy and anesthesia were not significant. Because NaPen did not alter secretion of LH or FSH after exogenous GnRH in experiment 1 while it blocked the postovariectomy increase in LH but not FSH in experiment 2, we concluded that the postovariectomy increase in LH resulted from increased hypothalamic secretion of GnRH. The mechanisms responsible for the postovariectomy increase in FSH secretion are not identical to those for LH. The mechanisms that control the postovariectomy secretion of FSH might involve factors that are not suppressible by NaPen or, alternatively, the differences in LH and FSH release after ovariectomy might reflect the removal of ovarian factors that suppress FSH but not LH secretion in intact ewes.     

Uterine influences on the formation of subnormal corpora lutea in seasonally anestrous ewes

The hypothesis that subnormal luteal function after induced ovulation in anestrous ewes was the result of uterine influences exerted during the periovulatory period was tested. Crossbred ewes (n = 27) in seasonal anestrus were induced to ovulate by administration of 12 doses of 250 ng of LHRH at 2-h intervals, followed immediately by a bolus injection of LHRH (250 micrograms; d 0). Ewes were unilaterally hysterectomized on either d -3 (PRELHRH) or 2 (POSTLHRH). Daily blood samples were collected and assayed for progesterone (P4) and 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM). All ewes were slaughtered on d 10, and corpora lutea (CL) were collected, weighed, and assayed for concentration of P4. All ewes that ovulated exclusively in the ovary ipsilateral to the remaining uterine horn had a transient increase in plasma P4 of 2 to 3 d (short luteal phase). In ewes with at least one CL in the isolated ovary, elevated plasma P4 was maintained after hysterectomy but was consistently lower (P less than .05) in POSTLHRH ewes than in PRELHRH ewes. Concentrations of PGFM did not differ between treatments. The CL ipsilateral to the remaining uterine horn weighted less (P less than .01) and contained less P4 (P less than .01) than contralateral CL. These data confirm the hypothesis that premature regression of subnormal CL is uterine-dependent in a local fashion. Presence of the uterus during the follicular and(or) early luteal phase inhibited subsequent luteal function in seasonally anestrous ewes.     

Androgen and estradiol effects on gonadotropin secretion and response to GnRH in ovariectomized pony mares

In Exp. 1, 16 long-term ovariectomized pony mares were used to determine the effects of treatment with estradiol benzoate (EB) and dihydrotestosterone (DHT) benzoate alone, and in combination, on secretion of follicle stimulating hormone (FSH) and luteinizing hormone (LH) in daily blood samples and after three consecutive injections of gonadotropin releasing hormone (GnRH). Administration of EB alone, or in combination with DHT, every other day for 11 d reduced (P less than .05) concentrations of FSH and increased (P less than .05) concentrations of LH in daily blood samples, and increased (P less than .05) the secretion of both gonadotropins after administration of GnRH. Treatment with DHT alone had no effect (P greater than .10) on LH or FSH concentrations in daily blood samples and no effect on the LH response to exogenous GnRH. There was no interaction (P greater than .10) between DHT and EB treatment for any hormonal characteristic. In Exp. 2, the control mares and mares treated with DHT in Exp. 1 were equally allotted to treatment with vehicle or testosterone propionate (TP) every other day for six injections, and then GnRH was administered as in Exp. 1. Treatment with TP had no effect (P greater than .10) on LH or FSH concentrations in daily blood samples but increased (P less than .05) the FSH response to exogenous GnRH, confirming our findings in previous experiments. It is concluded that the TP-induced stimulation of FSH secretion after exogenous GnRH in ovariectomized mares may involve estrogens produced from aromatization of the injected androgen.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of melatonin and progesterone administered to ewes in spring and summer

Melatonin (MEL) was evaluated for effects on LH, prolactin (PRL) and fertility in spring (Exp. 1, 2) and summer (Exp. 3 to 5). In Exp. 1, 17 ovariectomized ewes bearing estradiol implants were fed 3 mg MEL or vehicle for 44 d beginning May 1. Melatonin decreased (P less than .001) PRL levels but had no effect on LH secretion and response to GnRH. In Exp. 2, 12 ewes each received a 40-d MEL ear implant or a sham implant on March 31. Progesterone-releasing pessaries (CIDR) were applied for 12 d and were withdrawn concomitant with ram joining on May 7. Neither treatment stimulated follicular development or induced estrus or ovulation. Exp. 3 and 4 were contemporary 2 x 2 factorial trials with 24 ewes at each of two locations. Melatonin implants were administered on June 29 and CIDR on July 22. The CIDR were removed and rams (Exp. 3, vasectomized; Exp. 4, fertile) were joined on August 3. Days from introduction of rams to estrus were reduced (P less than .05) by CIDR but not by MEL. All ewes lambed in Exp. 4, and days to estrus and conception were reduced (P less than .001) by CIDR but not by MEL. Exp. 5 was designed like Exp. 4 except that MEL implants were inserted June 20 and rams were joined August 8. Intervals from introduction of rams to estrus were reduced (P less than .01) by both MEL and CIDR treatments.(ABSTRACT TRUNCATED AT 250 WORDS)     

Possible role of histamine in the regulation of secretion of luteinizing hormone in the ewe

Concentrations of histamine were quantified by an enzymatic isotopic assay in different regions of the brain and pituitary gland of gonadal-intact and chronically ovariectomized ewes during the anestrous season. Sera concentrations of LH were confirmed to be elevated in ovariectomized compared with intact animals immediately before tissues were obtained. Areas of the brain that were examined included cerebral cortex, thalamus, pineal gland, hypothalamus (rostral, medial basal, median eminence), midbrain, cerebellum and brain stem. Concentrations of histamine were greatest within the thalamus, pineal gland, medial basal hypothalamus and median eminence. Histamine within the medial basal hypothalamus was greater (P less than .05) in ovariectomized than in ovarian-intact animals. Further experiments were designed to determine the effect of antihistaminic drugs on secretion of LH. Ovariectomized ewes were treated every 6 h (i.m.) for 24 h with diphenhydramine (an antagonist of the H1-receptor for histamine), cimetidine (an H2-receptor antagonist), a combination of the drugs, or vehicle. Twelve hours after initiation of treatments, animals were injected with estradiol. Diphenhydramine depressed (P less than .01) basal serum concentrations of LH and the positive feedback effect of estradiol on serum concentrations of LH. Cimetidine did not influence the pattern of secretion of LH. Diphenhydramine did not alter LHRH-induced release of LH in ovariectomized ewes or basal serum concentrations of LH in ovarian-intact anestrous ewes. We suggest that histamine acts at the level of the central nervous system through an H1-receptor mechanism to control secretion of LH in female sheep.     

Naloxone-reversible inhibition of luteinizing hormone in postpartum ewes: effects of suckling and season

Involvement of endogenous opioids in inhibition of luteinizing hormone (LH) release and stimulation of prolactin (PRL) release was investigated by injecting the opioid antagonist naloxone into 18 ewes on d 7 and 8, d 12 and 13, and d 18 and 19 postpartum. Compared with control injections of saline, iv naloxone (1 mg/kg) increased serum concentrations of LH and decreased serum PRL in samples collected 15, 30 and 45 min after each injection. Ewes lambing in the spring (March) or autumn (September and October) that nursed one or two lambs did not differ in their LH and PRL responses to naloxone. Autumn-lambing ewes from which lambs were weaned within 1 d after parturition did not differ from ewes of the autumn-nursed group in any of the following characteristics: 1) serum LH increases following naloxone, 2) basal secretion of LH, 3) postpartum interval to first increase in serum progesterone and 4) relative decrease in serum PRL after naloxone despite large differences in basal PRL secretion. In summary, postpartum expression of a naloxone-reversible inhibition of LH release and stimulation of PRL secretion did not depend on suckling stimuli or differ between autumn and spring parturitions.     

Effect of cerebroventricular infusion of insulin and (or) glucose on hypothalamic expression of leptin receptor and pituitary secretion of LH in diet-restricted ewes

The objective was to determine the effect of central infusion of insulin and (or) glucose on hypothalamic expression of leptin receptor and pituitary secretion of LH in the ewe. Twenty-two ovariectomized ewes (32 wk of age) were fitted with two lateral cerebroventricular (LCV) cannulae and fed 33% of NRC requirements for 8 wk. Ewes (n> or =5/group) were then infused, via LCV cannulae, with artificial cerebrospinal fluid (aCSF) or aCSF containing physiological concentrations of insulin (INS), glucose (GLU), or INS + GLU; the mass of each increasing linearly from Day 0 (mass = 0 units/h) to Day 8 (mass of INS = 80 mIU/hr and GLU = 10 mg/hr). Jugular serum was collected every 12 min for 4 hr on Days 0, 2, and 4. Ewes treated with INS or INS + GLU had greater (P<0.06) mean concentrations of LH than aCSF treated ewes on Day 2 (13.8+/-1.8 and 12.5+/-1.3 > 8.0+/-3.3 ng/ml). Furthermore, on Day 4, concentrations of LH in INS treated ewes exceeded that (P<0.07) of aCSF treated ewes (14.8+/-2.0 > 7.4+/-3.0 ng/ml). Expression of NPY mRNA did not differ between treatments (P = 0.87). Leptin receptor mRNA expression was dramatically reduced (P<0.0002) in INS+GLU versus aCSF treated ewes. These data provide evidence to suggest that insulin may be an important component of hypothalamic mechanisms regulating secretion of LH and expression of leptin receptors in undernourished ruminants.     

Luteinizing hormone release after administration of the gonadotropin-releasing hormone agonist Fertilan (goserelin) for synchronization of ovulation in pigs

The generic GnRH agonist, Fertilan (goserelin), was tested for the ability to induce an LH surge and ovulation in estrus-synchronized gilts. Three experiments were performed to 1) examine the effect of various doses of Fertilan on secretion of LH in barrows, to select doses to investigate in gilts (Exp. 1); 2) determine doses of Fertilan that would induce a preovulatory-like rise of LH in gilts (Exp. 2); and 3) determine the time of ovulation after Fertilan treatment (Exp. 3). In Exp. 1, 10 barrows were injected on d 1, 4, 7, 10, and 13 with 10, 20, or 40 microg of Fertilan; 50 microg of Gonavet (depherelin; GnRH control) or saline (negative control); and sequential blood samples were collected for 480 min. There was a dose-dependent stimulation (P < 0.05) of LH release. Maximal plasma concentrations of LH (LH(MAX)) were 2.1 +/- 0.2, 4.1 +/- 0.3, 2.6 +/- 0.4, and 3.4 +/- 0.3 ng/mL after 10, 20, and 40 microg of Fertilan and 50 microg of Gonavet, respectively, and duration of release was 78 +/- 9, 177 +/- 12, 138 +/- 7, and 180 +/- 11 min, respectively. Fertilan doses of 10 and 20 microg were deemed to be the most suitable for testing in gilts. In Exp. 2, 12 gilts received (after estrus synchronization with Regumate and eCG) injections of 10 or 20 microg of Fertilan or 50 microg of Gonavet 80 h after eCG to stimulate a preovulatory-like LH surge and ovulation. An LH surge was induced in 3 of the 4 gilts in both of the Fertilan groups and in all of the Gonavet-treated gilts. Characteristics of induced release of LH did not differ among groups: LH(MAX), 5.0 +/- 0.9 vs. 4.6 +/- 1.8 vs. 6.6 +/- 1.1 ng/mL; duration, 11.7 +/- 2.0 vs. 12.3 +/- 2.2 vs. 14.3 +/- 0.5 h; interval from GnRH injection to LH(MAX), 4.0 +/- 2.0 vs. 6.7 +/- 1.3 vs. 5.8 +/- 1.6 h. In Exp. 3, estrus-synchronized gilts were injected with 20 microg of Fertilan (n = 8) or 50 microg of Gonavet (n = 4), and the time of ovulation was determined by repeated endoscopic examination. Time of ovulation ranged from 34 to 42 h postGnRH; however, ovulation occurred earlier in the Gonavet compared with the other groups (P < 0.05). Results of these experiments indicate that 1) barrows are an appropriate model for determining GnRH doses that can be effective in inducing a preovulatory-like LH surge in females; 2) the generic GnRH agonist Fertilan, at doses of 10 to 20 microg, can stimulate an LH surge in gilts, with subsequent ovulation; and 3) Fertilan at doses of 10 and 20 microg should be examined further for use in fixed-time insemination protocols.     

Endogenous opioid modulation of luteinizing hormone and prolactin secretion in postpartum ewes and cows

A possible role for endogenous opioid peptides (EOP) in the control of luteinizing hormone (LH) and prolactin (PRL) secretion was studied by injecting the opioid antagonist, naloxone (NAL), into postpartum ewes and cows. Twelve ewes that lambed during the fall breeding season and nursed their lambs were injected iv with NAL (1.0 mg/kg) on d 10, 14, 18, 22 and 26 postpartum. Blood samples were collected at 15-min intervals from 2 h before to 2 h after NAL, and serum concentrations of LH and PRL were quantified. Following treatment on d 10, suckling lambs were removed from 6 of the 12 ewes, creating non-suckled (NS) and suckled (S) treatment groups for subsequent study on d 14 through 26. On d 10, NAL treatment increased LH (P less than .01) but concentrations of PRL were not affected. When averaged across d 14 to 26, post-NAL concentrations of LH were greater (P less than .001) than pre-NAL concentrations (6.5 +/- .7 vs 1.9 +/- .4 ng/ml). In contrast, concentrations of PRL in the post-NAL period were lower (P less than .001) than pre-NAL concentrations (129 +/- 15 vs 89 +/- 10 ng/ml). Compared with S ewes over d 14 to 26, those in the NS group had similar pre-NAL concentrations of LH, tendencies for higher (P less than .10) post-NAL concentrations of LH, lower (P less than .001) mean serum concentrations of PRL (pre- and post-NAL) and similar pre-NAL vs post-NAL differences in serum PRL. Six suckled beef cows on d 24 to 35 were injected iv with either saline or NAL (.5 mg/kg) in a replicated crossover design. Injections of NAL increased serum concentrations of LH (P less than .05), when averaged over all 12 injections in the six cows, but serum PRL was not changed. However, three of six cows did not respond to NAL with increases in serum LH. These non-responding cows were similar to the responding cows in their pre-injection concentrations of LH and PRL, but they tended (P = .10) to have higher serum concentrations of cortisol than responding cows.     

Changes in metabolic and reproductive characteristics associated with lactation and glucose infusion in the postpartum ewe

This study examined mechanisms whereby the metabolic environment interacts with basic reproductive function. Ewes lambing during the breeding season were fed to maintain (MAINT, n = 10) or gain (GAIN, n = 11) body weight during the last 4 mo of gestation. From d 7 to 22 postpartum, ewes were infused iv with saline (n = 10) or glucose at a rate calculated to increase normal glucose entry rate by 75% (n = 11). Blood samples were collected daily to determine plasma concentrations of nutritive metabolites and insulin and at frequent intervals on d 14 and 21 to determine serum gonadotropin concentrations. Hypothalami and pituitaries were collected on d 22 to determine hormone content and receptor concentrations. Plasma concentrations of nutritive metabolites and insulin indicated that MAINT ewes mobilized more (P less than .01) body fat and protein reserves during gestation and early lactation than ewes in the GAIN group. Glucose infusion elevated plasma concentrations of glucose (P less than .05) and insulin (P less than .07) and reduced (P less than .05) fat and protein mobilization, even though it depressed feed intake (P less than .001), compared with saline infusion. Hypothalamic gonadotropin-releasing hormone (GnRH), pituitary luteinizing hormone (LH) and follicle-stimulating hormone (FSH) content and pituitary GnRH receptor concentration were similar between treatments.(ABSTRACT TRUNCATED AT 250 WORDS)     

Gonadotropin releasing hormone-induced secretion of luteinizing hormone during the milk-ejection reflex in the postpartum beef cow

A study was conducted to determine the effect of the milk-ejection reflex on exogenous gonadotropin releasing hormone (GnRH)-induced release of luteinizing hormone (LH) after short-term calf removal. Twenty-four postpartum multiparous beef cows were assigned randomly to groups arranged in a 2(3) factorial arrangement. Factors consisted of two levels of suckling [suckled (S) or nonsuckled (NS)], treatment with GnRH [saline (C) or 200 micrograms GnRH] and days postpartum (d 1 and 14). Dams were isolated from their calves for 4 h on d 1 and 14 postpartum. At the end of 4 h dams were reunited with their calves in S + C and S + GnRH groups, while dams of calves in NS + C and NS + GnRH groups remained separated an additional 2 h. Cows were injected iv with saline or GnRH following the 4-h isolation period, 5 min after calves had begun suckling or nuzzling the udder. Sera from jugular blood samples collected 15 min prior to the end of the 4-h isolation period, immediately prior to injection (0 h) and at 15 min intervals thereafter for 120 min were analyzed for LH. Serum concentrations of LH in control cows did not differ due to suckling or stage of the postpartum period and averaged 2.3 +/- .1 ng/ml. Pituitary response to GnRH was determined by computing the rate of LH release. Rate of LH release (ng LH.ml-1.min-1) in response to GnRH on d 14 was greater (P less than .001) than on d 1 in both suckled and nonsuckled groups (S + GnRH, 37.1 +/- 3.9 vs 18.3 +/- 5.0; NS + GnRH, 34.7 +/- 5.9 vs 14.5 +/- 1.1). However, GnRH-induced release of LH did not differ between suckled and nonsuckled cows on either d 1 or 14 postpartum. These data indicate that response of the bovine pituitary to GnRH during the postpartum period is not influenced by the act of suckling but is enhanced with time after parturition.