Ovarian function and hormone secretion of gilts actively immunized against androstenedione

Fifty crossbred gilts immunized against bovine serum albumin (BSA) or androstenedione conjugated to BSA (AD) were used in three experiments. Primary immunizations were given at 120 d of age and boosters at 148 and 176 d. Gilts were moved to pens containing four to five animals each and exposed to boars beginning at 180 d of age. Immunization against AD did not affect age at puberty, percentage of gilts exhibiting estrus or duration of first estrous cycle. Over the three experiments, ovulation rate was 24% greater for AD-immunized gilts than for controls, and the number of corpora lutea was related positively (r = .82) to the log of the antibody titer. Number of ovulations decreased as interval from booster immunization to onset of estrus increased. During diestrus of the first estrous cycle, gilts immunized against AD had more follicles 5 to 10 mm in diameter, more total ovarian follicles and more total ovarian structures (corpora lutea plus follicles) than controls. Immunization against AD increased the frequency of LH pulses on d 16 but not on d 17 or 18, of the estrous cycle. However, average serum concentrations of LH, FSH and estradiol from 5 d before until 2 d after expected estrus were not different between treatment groups. Concentrations of AD in follicles 4 to 6 and greater than 7 mm in diameter were greater in gilts immunized against AD. Mean serum progesterone was higher on d 9 and 12 after mating in AD immunized gilts than in controls. Immunization against AD had no effect on maintenance of pregnancy or embryo survival rate.     

Ovarian response of the hypophysial stalk-transected pig to pregnant mare serum gonadotropin

Hypophysial stalk transection (HST) or sham operation (S-HST) was performed on 14 prepuberal gilts, 169 ± 3 days of age and 72.8 ± 3.4 kg body weight (day of surgery = Day 0). Gilts received 1,000 IU of pregnant mare's serum gonadotropin (PMSG) or saline vehicle (V) intramuscularly (im) on Day 2 resulting in the following groups: S-HST + V (n=3), S-HST + PMSG (n=4), HST + V (n=3) and HST + PMSG (n=4). Ovarian morphology and weights were recorded after ovariectomy on Day 6. Gilts were weighed on Day 59 ± 3. Sequential blood samples were collected via jugular vein cannula from 6 S-HST and 6 HST gilts on Day 60 ± 3, and all gilts were necropsied on Day 86 ± 7. Body weight gain and whole pituitary gland weight were greater (P<0.05) for S-HST than HST gilts. Mean serum LH concentration, basal serum LH concentration, frequency of LH peaks and LH peak amplitude were greater (P<0.005) for S-HST than HST gilts. Serum PRL and GH, were similar for both groups. Total ovarian and follicular fluid weights were greater (P<0.05) in S-HST gilts given PMSG than those of the other three groups which did not differ. Therefore, PMSG stimulated follicular growth in S-HST gilts, but failed to stimulate follicular growth in HST gilts. We suggest that a critical basal serum LH and/or FSH concentration must be maintained to support and promote follicular growth and a pulsatile delivery of GnRH to the anterior pituitary gland by an intact brain-pituitary unit may be required to provide this basal serum gonadotropin concentration.     

Pulsatile administration of gonadotropin-releasing hormone agonist to gilts actively immunized against gonadotropin-releasing hormone

Sexually mature gilts (n = 20) were actively immunized against GnRH. Primary and booster immunizations of GnRH conjugated to bovine serum albumin induced production of antibodies in all gilts. Nineteen of the gilts became acyclic with suppressed concentrations of gonadotropins and estradiol. Intravenous challenges with 100 micrograms GnRH and 5 micrograms D-(Ala6, des-Gly-NH2(10)) ethylamide GnRH (a GnRH agonist that did not cross-react with antibodies produced by the gilts) caused release of LH and FSH, indicating maintenance of secretory capacity of pituitary gonadotropes in the immunized animals. Gilts were given 100 ng GnRH agonist at 2-h intervals for 72 h (n = 4) or 144 h (n = 10) or did not receive agonist (n = 5). Blood samples were taken every 6 h, and detectable concentrations of LH were observed in 42% and 52% of samples taken from gilts treated with or without agonist. In contrast, serum concentrations of FSH and estradiol were undetectable. Reproductive tracts and anterior pituitaries were taken from gilts at the conclusion of pulsatile administration of GnRH agonist or at 144 h for controls. Pituitary concentration of LH and FSH, uterine wet and dry weight, and size of the uterus were similar among groups. Paired ovarian weights for treated gilts pulsed with GnRH agonist for 72 h were heavier (P less than .05); however, ovaries from all immunized gilts were atrophied without follicular structures.(ABSTRACT TRUNCATED AT 250 WORDS)     

Response of gilts with delayed puberty to pregnant mare serum gonadotropin or estrogen

The effect of pregnant mare serum gonadotropin (PMSG) or estradiol cyclopentylpropionate (EC) on the induction of estrus, duration of estrus, and serum progesterone concentration after estrus was evaluated in 8 gilts with delayed puberty. Four gilts were given 500 IU of PMSG IM and 4 were given 2 mg of EC, IM. The inactive status of the ovaries at the time of treatment was verified by serum progesterone values of less than 0.5 ng/ml in serial samples collected before treatment. The 4 EC-treated gilts came into estrus at a mean of 3.5 days after treatment, but 1 of the gilts did not form corpora lutea. Three PMSG-treated gilts came into estrus at a mean of 4.0 days after treatment. The remaining PMSG-treated gilt remained anestrus and did not form corpora lutea. The mean duration of estrus in EC-treated gilts was 5.25 days compared with 2.0 days for PMSG-treated gilts (P less than 0.05). Serum progesterone concentrations were higher in PMSG-treated gilts than in EC-treated gilts at 8, 11, and 17 days after treatment (P less than 0.05).     

Luteinizing hormone (LH) response to different doses of synthetic gonadotropin releasing hormone (GnRH) in prepubertal gilts

The object of this investigation was to study luteinizing hormone (LH) response to different doses of synthetic gonadotropin-releasing hormone (GnRH) in prepubertal gilts. Four crossbred prepubertal gilts, 128–134 days old and body weight 57–63 kg, were used in this study. Four doses, 0. 5, 25 and 125 μg, of GnRH were administered via a jugular vein catheter in a latin square design. Each treatment consisted of 3 injections at 90 min intervals. Frequent blood samples were taken during a period of 90 min before up to 90 min after treatment. Total LH responses were measured from post-treatment samples as the area under the curve above base level obtained from pre-treatment samples. A positive relationship between GnRH dose and LH release was obtained in all gilts, except for 1 treatment given to a gilt with high plasma level of oestradiol-17β on the day of treatment. This study has demonstrated the responsiveness of the pituitary gland by LH release to different doses of GnRH in 4.5-month-old prepubertal gilts.     

Ovulation and embryonic survival in pubertal gilts treated with gonadotropin releasing hormone

An experiment was conducted to evaluate the effect of exogenous gonadotropin releasing hormone (GnRH) on ovulation and embryonic survival in pubertal gilts. Gilts were assigned in replicates to a control (n = 10) and treatment (n = 10) group. Treatment consisted of an iv injection of 200 micrograms of GnRH immediately after initial mating on the first day of detected estrus. Control gilts were similarly injected with physiological saline. Blood samples were collected from the anterior vena cava immediately prior to injection, thereafter at 15-min intervals for 90 min, and subsequently, before slaughter on d 30 of gestation. Serum samples were analyzed for luteinizing hormone (LH) and progesterone by radioimmunoassay. Treatment with GnRH increased the quantity of LH released (P less than .05), with highest serum concentrations (ng/ml, means +/- SE) of gonadotropin in treated gilts (17.3 +/- 3.5) occurring at 75 min post-injection. In control gilts, serum concentrations of LH were not affected by injection of saline. Mean number of ovulations in treated gilts was also greater (P less than .05) than that of control animals (14.5 +/- .7 vs 12.1 +/- .6). However, treatment with GnRH did not enhance the number of attached conceptuses (normal and degenerating) present (treated, 10.9 +/- .9 vs control, 10.5 +/- .7) nor the percentage of viable fetuses (treated, 74.7 +/- 6.9 vs control, 83.5 +/- 5.0%) on d 30 of gestation. Although GnRH increased ovulation rate, mean weight of corpora lutea of treated and control gilts did not differ (402.8 +/- 16.3 vs 389.5 +/- 11.3 mg, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)     

Reproductive and growth responses of gilts to exogenous porcine pituitary growth hormone

Forty gilts (mean wt = 72 kg) were administered daily either vehicle (C = control) or 70 micrograms porcine growth hormone (pGH)/kg BW. After 30 d of treatment, eight gilts per group (Exp. 1) were slaughtered and blood, uteri and ovaries were collected. Follicular fluid (FFl) was collected and granulosa cells (GC) were cultured. The remaining gilts (Exp. 2) were treated for up to 35 additional days and examined twice daily for estrus. Estrusal gilts were removed from the experiment. Noncyclic gilts (n = 9 of 12 pGH; n = 4 of 12 C) were slaughtered on d 66 and their ovaries were examined. Ovarian weights were not different for pGH and C gilts in either Exp. 1 (P greater than .1) or Exp. 2 (P = .09). Uterine weights were greater for pGH-treated than for C gilts (P less than .007) in Exp. 1, but not in Exp. 2. Concentrations of estradiol (E2) in plasma and FF1 and of progesterone (P) in plasma and FF1 were not different for pGH and C gilts. Concentrations of insulin-like growth factor-I (IGF-I) in FF1 and in serum were greater for pGH than for C gilts (P less than .01). Concentration of P in serum-free medium of cultured GC was lower for GH than for C (P less than .05) in the presence or absence of gonadotropins in Exp. 1. The FSH-stimulated secretion of P was also lower for GC of pGH-treated gilts in Exp. 2, indicating a failure of GC to differentiate in culture. Only one pGH gilts in Exp. 2 manifested estrus, compared with seven C gilts (P less than .025). In Exp. 1, ADG was higher (P less than .03) and feed/gain lower (P less than .07) for pGH gilts. Longissimus muscle area (LMA) was not different (P = .19) between groups. Backfat thickness (BF) was lower (P less than .005) in pGH than in C in both Exp. 1 and 2. We conclude that exogenous pGH increased growth rate, improved feed efficiency and altered carcass traits in gilts. However, these effects were associated with impaired ovarian development of prepubertal gilts and a low incidence of estrus.     

Aggressive behavior is reduced in bulls actively immunized against gonadotropin-releasing hormone

The purpose of this research was to compare the frequency of aggressive behavior's in beef bulls actively immunized against gonadotropin-releasing hormone relative to contemporary nonimmunized control bulls and surgically castrated steers. Eight males were assigned to each ofthese treatments in each of 4 yr. Immunized males were treated with a GnRH-keyhole-limpet hemocyanin (KLH) conjugate at approximately 4 mo of age. A secondary (booster) immunization was administered at 12 mo. Steers were castrated at 4 mo of age. Animals in each treatment in each year were housed as a single group prior to testing. At approximately 16 mo of age, each group of eight animals was placed in a 10- x 16-m enclosure for 20 min on five occasions at 2 to 3 d intervals. An observer recorded butts initiated by each animal as well as participation in bouts of sparring. Relative to control bulls, immunocastration reduced the frequency of butts initiated (P < 0.05) and participation in sparring bouts (P < 0.05) to levels typically observed in steers (P > 0.05). These observations indicate that active immunization against GnRH reduces the incidence of aggressive behavior in male beef cattle and are consistent with our postulate that immunoneutralization of GnRH is an effective alternative to surgical castration in the management of beef cattle.     

Serum prolactin response to thyrotropin releasing hormone in estrogen treated hypophysial stalk-transected gilts

Twelve crossbred gilts, 169 ± 3 days of age and 72.8 ± 3.4 kg body weight, were hypophysial stalk-transected (HST)1 or sham hypophysial stalk-transected (S-HST). Gilts were ovariectomized 6 days later and assigned to four treatments of 3 gilts each in a 2 × 2 factorial arrangement. One-half of the HST and S-HST gilts received 5 mg estradiolbenzoate (EB) or corn oil vehicle im at 0800 hr daily for 5 days beginning 64 ± 3 days after HST or S-HST. Blood was collected by jugular vein cannula at 0830 and 0900 hr the day after the last injection of EB or oil. Immediately after the 0900 hr sample, 200 μg thyrotropin releasing hormone (TRH) were injected (iv). Mean basal serum prolactin (PRL) concentration was similar for HST (10.3 ± 1.0 ng/ml) and S-HST (12.3 ± 1.7 ng/ml) gilts, however mean basal serum PRL concentration was greater (P<.05) for EB-treated gilts (13.7 ± 1.3 ng/ml) than for oil-treated gilts (8.8 ± .5 ng/ml). Mean serum PRL concentration of all gilts increased within 10 min and returned to approximately 20 ng/ml by 150 min after TRH. Maximum serum PRL concentrations at 10 min after TRH were greater (P<.01) for S-HST (255.9 ± 29.6 ng/ml) than HST gilts (83.4 ± 18.8 ng/ml), but were not different for EB (198.0 ± 50.6 ng/ml) and oil-treated gilts (141.4 ± 36.3 ng/ml). Area under the serum PRL response curve after TRH was greater (P<.005) for S-HST than HST gilts and for EB than oil-treated gilts (P<.05). These results do not eliminate the possible influence of estrogen on PRL secretion at the hypothalamus, but do indicate that estrogen directly stimulated the anterior pituitary gland to secrete PRL.     

Effects of pregnant mare serum gonadotropin (PMSG) on pituitary responsiveness to progesterone and LHRH in the hen,Gallus domesticus

Laying hens were primed once daily with either pregnant mare serum gonadotropin (PMSG) or saline until spontaneous ovulations were suppressed (PMSG-treated birds) or for 5–7 days (controls). PMSG-primed hens and saline-primed hens were subsequently injected with vehicle, 500 μg progesterone or 20 μg [Arg⁸]-LHRH. Twelve hr following the injection, PMSG-treated hens were found to lack a normal follicular hierarchy and to have a significantly greater number of follicles larger than 2.5 cm compared to saline-primed hens. Treatment with PMSG also increased basal plasma concentrations of estradiol-17β and decreased plasma LH compared to controls. Finally, challenge with progesterone or LHRH failed to either stimulate LH release or to overcome the ovulation-suppressing effect of PMSG. Results suggest that the action of PMSG in the hen can be attributed, at least in part, to the unresponsiveness of the pituitary to the normal LH-stimulating hormone(s).     

Feedlot performance of steers and bulls actively immunized against gonadotropin-releasing hormone.

Feedlot performance and testicular and pituitary function were assessed in cattle actively immunized against GnRH. In Trial 1, 50 steers were either unimmunized (n = 10), actively immunized against keyhole limpet hemocyanin (KLH; n = 10), or immunized against a GnRH-KLH conjugate (n = 30). Fifteen of 30 steers immunized against GnRH-KLH received a secondary immunization 8 wk after primary immunization. Antibodies against GnRH were not evident in unimmunized steers or steers actively immunized against KLH. Antibodies against GnRH were noted in all immunized animals (n = 30) within 6 wk of primary immunization and anti-GnRH antibody concentrations became maximal 20 to 24 wk after immunization. The increasing anti-GnRH titer in immunized steers was associated with decreasing serum concentrations of LH. Serum concentrations of LH were depressed (P less than .05) within 8 wk of primary immunization and reached a nadir by wk 20. The patterns of increase in GnRH titer and decrease in serum concentrations of LH did not differ (P greater than .05) in animals receiving primary immunization alone or primary and secondary immunization. Feedlot performance and carcass quality were not affected (P greater than .05) by immunization against KLH or the GnRH-KLH conjugate. In Trial 2, 60 bull calves (mean weight = 325.2 +/- 2.8 kg) were randomly assigned to a 2 x 3 factorial experiment. The two classes (n = 30) were 1) unimplanted and 2) implanted with Synovex-S. The three treatments (n = 20) were 1) intact control, 2) actively immunized against GnRH, and 3) castrate.(ABSTRACT TRUNCATED AT 250 WORDS)     

Sexual dimorphic porcine pituitary response to growth hormone-releasing hormone

The aim of this study was to compare growth hormone (GH) response of barrows and gilts to porcine growth hormone-releasing hormone (pGRH) at the pituitary level. Anterior pituitary cells from barrows and gilts responded to pGRH in a dose-dependent manner. The median effective pGRH concentration (EC50) which stimulated GH release from cells of barrows was greater (P less than .05) than that for cells obtained from intact female siblings. Maximal pGRH mediated GH secretion from barrows was not different (P greater than 0.05) than that from gilt stimulated cells. These data demonstrate that somatotrophs of growing peripubertal gilts are more responsive to pGRH stimulation than are cells from their castrated male siblings. This difference could be caused by castration of the neonatal male.     

Follicle growth in hypophysial stalk-transected pigs given pulsatile GnRH and pregnant mare serum gonadotropin

In experiment 1, nine prepuberal crossbred gilts 145 +/- 2 days of age and 90.3 +/- 1.6 kg body weight (BW) were hypophysial stalk-transected (HST) or sham-HST. Starting at 0800 on Day 1 (35 +/- 2 days after surgery), three sham-HST and two HST gilts received 3.5% sodium citrate vehicle (V) while two HST gilts and two sham-HST gilts received pulses of 2.5 micrograms GnRH every 45 min for 9 days via a jugular vein cannula. At 0800 on day 7, all gilts received 1,000 IU of pregnant mare serum gonadotropin (PMSG) im. Blood was sampled every 15 min from 0800 to 0845 on Days 1 through 6. On Day 10, ovarian morphology and ovarian and follicular fluid weights were recorded. In experiment 2, eight prepuberal crossbred gilts, 146 +/- 6 days of age and 79.5 +/- 1.5 kg BW, were HST or sham-HST. Starting at 0800 on Day 1 (7 +/- 4 days after surgery), two sham-HST and three HST gilts received V, while three HST gilts received pulses of 2.5 micrograms GnRH every 45 min for 8 days. At 1200 on Day 5, all gilts, including three unoperated controls (UC), received 1,000 IU of PMSG im. Blood was sampled from all but UC gilts every 15 min from 0800 to 0845 on Days 1 through 5. Ovarian data were obtained on Day 9. The HST + V gilts failed to respond to PMSG, whereas growth of ovulatory follicles was stimulated in the other groups in both experiments.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of feed restriction on serum somatotropin, insulin-like growth factor-I-(IGF-I) and IGF binding proteins in cyclic heifers actively immunized against growth hormone releasing factor

Feed restriction often increases serum somatotropin (ST) and decreases insulin-like growth factor-I (IGF-I) in ruminants; however, the mechanisms responsible for this change in ST and IGF-I are not well defined. We investigated the effects of feed restriction on serum ST, IGF-I, IGF binding proteins (IGFBP), insulin and nonesterified fatty acids (NEFA) in cyclic Angus and Charolais heifers (n=15) previously immunized against growth hormone releasing factor (GRFi) or human serum albumin (HSAi). Cows were fed a concentrate diet ad libitum (AL) or were restricted to 2 kg cotton seed hulls (R) for 4 d. Each heifer received each dietary treatment in a single reversal design. As anticipated, GRFi decreased ST, IGF-I and insulin (P<.05). In addition, GRFi decreased serum IGFBP-3 (P<.01), but increased IGFBP-2 (P<.01). Feed restriction resulted in an increase in serum ST in HSAi, but not in GRFi heifers. Regardless of immunization treatment, feed restriction decreased serum IGF-I and insulin, and increased NEFA (P<.01). In conclusion, the increase in serum ST levels observed during feed restriction was blocked by active immunization against GRF. However, feed restriction resulted in decreased serum IGF-I in GRFi heifers in spite of initial low levels of IGF-I (due to GRFi). Although GRFi decreased levels of IGFBP-3 and increased levels of IGFBP-2, feed restriction for 4 d did not alter serum IGFBP.     

Increased luteinizing hormone secretion and ovarian function in heifers actively immunized against estrogen and progesterone

This study was designed to test the effects of active immunization against estrogen and progesterone on patterns of luteinizing hormone (LH) and follicle stimulating hormone (FSH) secretion, ovarian characteristics and growth rate of heifers. Heifers were randomly assigned to four treatments: 1) control injection (n = 10); 2) ovariectomy (n = 9); 3) immunization against estrogen (anti-E, n = 10); and 4) immunization against estrogen and progesterone (anti-E+P4, n = 10). Three booster immunizations were administered at 1, 1.5 and 6 mo after primary immunization. Progesterone antibody binding was 40% (34 fmol at 1:600 final dilution) in the anti-E+P4 heifers, and estradiol-17 beta binding was 35% (30 fmol) and 60% (52 fmol at 1:100 final dilution) in the anti-E+P4 and anti-E heifers, respectively, after the final immunization. Anti-E+P4 heifers had more pulses of LH and higher basal concentrations of LH than anti-E or control heifers (P less than .05). Concentrations of LH in anti-E+P4 heifers did not increase to concentrations found in ovariectomized heifers (P less than .05). Immunization against steroids did not alter the secretion of FSH. The number of large follicles (greater than 15 mm diameter) in anti-E+P4 and anti-E heifers was greater than in control heifers (P less than .05). Ovarian weight was increased in anti-E+P4 heifers (P less than .05). Average daily gain was not different among groups (P greater than .05). It was concluded that active immunization against estrogen and progesterone in heifers increased LH secretion and stimulated ovarian function.     

Effect of unilateral castration and unilateral cryptorchidism on gonadotropin and testosterone response to gonadotropin releasing hormone in the bull

The effects of unilateral castration (UC) and induced unilateral cryptorchidism (UCR) on basal plasma luteinizing hormone (LH), follicle stimulating hormone (FSH) and testosterone, and on the responses of these hormones to gonadotropin releasing hormone (GnRH), were investigated in bulls altered at 3, 6 or 9 months of age. Blood plasma was collected before and after GnRH (200 micrograms) stimulation approximately 1 year following gonadal manipulation. Neither mean baseline concentrations nor GnRH-induced increases in plasma testosterone were altered (P greater than .1) by hemicastration or UCR (P greater than .1). Both mean baseline LH and GnRH-induced LH release were greater (P less than .05) in bulls altered at 3 months of age than in bulls altered at 9 months of age. UC increased (P less than .05) plasma LH response to GnRH over that observed in intact bulls, but not above that in UCR bulls. UCR had no detectable effect on either baseline concentrations or GnRH-stimulated LH release. FSH was increased (P less than .05) in hemicastrates, while UCR had a variable effect on peripheral FSH: FSH was reduced (P less than .05) in UCR animals altered at 3 months of age but increased (P less than .05) in UCR bulls altered at both 6 and 9 months of age when compared to FSH in intact bulls. The results indicate that, compared with intact bulls, UC bulls release increased amounts of both gonadotropins but similar amounts of testosterone in response to GnRH stimulation. UCR had a variable effect on FSH release and did not alter either LH or testosterone.     

Immunization of sheep against modified peptides of gonadotropin releasing hormone conjugated to carriers

The efficacy of antigens based on modified GnRH peptides in stimulating the production of antibodies against GnRH in sheep was tested. In the first study cysteine-containing GnRH peptides were conjugated to keyhole limpet haemocyanin (KLH) in 3 different orientations. The 3 conjugates were prepared in an emulsion of Freund's complete adjuvant (FCA) and were injected into 3 groups of 6 castrated male lambs. The 3 vaccines efficiently induced anti-GnRH titers in all the animals treated. The specificity of the GnRH antisera raised varied depending on the orientation of the GnRH molecule in the antigen and on the individual animal.

In a second trial designed to evaluate carrier molecules, a cysteine-containing GnRH peptide was conjugated to either KLH, equine serum albumin, ovalbumin or tetanus toxoid. The conjugates were prepared with FCA and injected into intact male lambs. All 4 vaccines stimulated the production of antibodies against GnRH in all the animals treated. The conjugates prepared with equine serum albumin or ovalbumin were the most effective in raising high anti-GnRH titers. In 18 of 20 lambs treated, anti-GnRH titers resulted in a marked atrophy of the testes.

We conclude that: 1) the different epitopes of the GnRH molecule are equally immunogenic in sheep; 2) the GnRH antibody response is affected by the carrier used; and, 3) anti-GnRH vaccines based on cysteine-substituted GnRH analogues show potential for use in immunocastration of livestock.     

Effects of active immunization against gonadotropin releasing hormone on gonadotropin secretion after ovariectomy and testosterone propionate administration to mares

Five lighthorse mares were actively immunized against gonadotropin releasing hormone (GnRH) conjugated to bovine serum albumin (BSA) to study the involvement of GnRH in luteinizing hormone (LH) and follicle stimulating hormone (FSH) secretion following ovariectomy (OVX) and after administration of testosterone propionate (TP). Five mares immunized against BSA served as controls. Immunizations were started on November 1, and OVX was performed in June (d 1). All mares were treated with TP from d 50 to 59 after OVX. On the day of OVX, concentrations of LH were lower (P less than .05) in GnRH-immunized mares than in BSA-immunized mares and were generally nondetectable; FSH concentrations were reduced (P less than .05) by 50% in GnRH-immunized mares relative to BSA-immunized mares. In contrast to BSA-immunized mares, plasma concentrations of LH or FSH did not increase after OVX in GnRH-immunized mares. The LH response to GnRH analog (less than .1% cross-reactive with GnRH antibodies) on d 50 was reduced (P less than .05) by 97% in GnRH-immunized mares relative to BSA-immunized mares, whereas the FSH response was similar for both groups. Treatment with TP for 10 d reduced (P less than .01) the LH response and increased (P less than .01) the FSH response to GnRH analog in BSA-immunized mares, but it had no effect (P greater than .1) on the response of either gonadotropin in GnRH-immunized mares.(ABSTRACT TRUNCATED AT 250 WORDS)