Lamprey gonadotropin-releasing hormone-III selectively releases follicle stimulating hormone in the bovine

Recent studies have shown that lamprey gonadotropin-releasing hormone (l-GnRH) is localized in the mammalian brain, and that l-GnRH-III, can selectively induce FSH secretion in the rat both in vivo and in vitro. Consequently, the purpose of this study was to determine if l-GnRH-III could elicit selective FSH release in cattle and compare this response with that to mammalian luteinizing hormone releasing hormone (m-LHRH). Cattle were chosen as the animal model because previous studies have demonstrated that FSH and LH are secreted by separate gonadotropes in that species. For these studies, crossbred cycling heifers were implanted with jugular cannulae and l-GnRH-III was infused either between Days 9–14 or on Day 20 of the estrous cycle. Blood samples were collected both before and following peptide infusion. Our results demonstrate that during Days 9–14 of the estrous cycle (luteal phase), when progesterone levels averaged between 4 and 5 ng/ml, a dose of 0.25 mg of l-GnRH-III induced the release of FSH (P < 0.05), but not LH. A 0.5 mg dose of l-GnRH-III caused a greater release of FSH (P < 0.01), but still did not induce LH release. Higher doses of the peptide were capable of significantly releasing both gonadotropins. Importantly, during the luteal phase, doses of 0.5 and 2 mg of m-LHRH were ineffective in stimulating FSH, but did elicit marked increases (P < 0.001) in LH. Again, progesterone levels averaged 4–5 pg/ml. In order to assess gonadotropin releasing ability of l-GnRH-III at a different phase of the estrous cycle, some animals were administered the peptide on Day 20, when progesterone levels were below 1.0 pg/ml. At this time, the l-GnRH-III induced the release of LH (P < 0.01), but not FSH. Overall, our results demonstrate that l-GnRH-III can selectively induce FSH in cattle during the luteal phase, whereas m-LHRH was ineffective in that regard. Furthermore, the fact that l-GnRH-III can selectively stimulate FSH when serum progesterone is high, and LH when serum progesterone is low, suggests its actions are under strong control of this steroid. We suggest the FSH releasing capacity of l-GnRH-III in cattle could render this peptide useful for enhancement of reproductive efficiency in this species.     

Evaluation of steroidogenic capacity after follicle stimulating hormone stimulation in bovine granulosa cells of Revalor 200~ implanted heifers

Background： Heifers not used as breeding stock are often implanted with steroids to increase growth efficiency thereby altering hormone profiles and potentially changing the environment in which ovarian follicles develop. Because bovine granulosa cell culture is a commonly used technique and often bovine ovaries are collected from abattoirs with no record of implant status, the objective of this study was to determine if the presence of an implant during bovine granulosa cell development impacts follicle stimulating hormone-regulated steroidogenic enzyme expression. Paired ovaries were collected from 16 feedlot heifers subjected to 1 of 3 treatments： non-implanted （n = 5）, Revalor 200 for 28 d （n = 5）, or Revalor 200 for 84 d （n = 6）. Small follicle （1 to 5 mm） granulosa cells were isolated from each pair and incubated with phosphate buffered saline （n = 16） or 100 ng/mL follicle stimulating hormone （n = 16） for 24 h. Results： Granulosa cells of implanted heifers treated with follicle stimulating hormone produced medium concentrations of progesterone similar （P = 0.22） to non-implanted heifers, while medium estradiol concentrations were increased （P 〈 0.10） at 28 and 84 d compared to non-implanted heifers indicating efficacy of treatment. Additionally, real-time PCR analysis in response to follicle stimulating hormone treatment demonstrated a decrease in steroidogenic acute regulatory protein （P = 0.05） mRNA expression in heifers implanted for 84 d and an increase in P450 side chain cleavage mRNA in granulosa cells of heifers implanted for 28 （P 〈 0.10） or 84 d （P 〈 0.05） compared to non-implanted females. However, no difference in expression of 3-beta-hydroxysteroid dehydrogenase （P= 0.57） and aromatase （P = 0.23） were demonstrated in implanted or non-implanted heifers. Conclusions： These results indicate follicles which develop in the presence of high concentrations of androgenic and estrogenic steroids via an implant tend to demonstrate an alter     

促卵泡素在奶牛超排效果上的对比试验

奶牛是单胎动物,超数排卵是有效进行奶牛胚胎移植所必需的技术环节,也是胚胎移植中最难控制的一个环节。一般来说平均每个供体牛可提供5-6个可用胚胎。虽然一头供体牛有时可提供20枚甚至更多的受精卵,但很多时候会得不到l枚可移植的胚胎。影响超排效果的因素主要有两方面,一方面是个体差异导致不同的超排反应;另一方面,不同厂家生产的促卵泡素（FSH）制剂成份比例不同,也是影响胚胎质量和数量的重要因素。目前国产的FSH制品均不是提纯品,而且含有两种活性成分,即FSH／LH。其比例常因厂家和批号不同而有较大的变化。在2005年4月至7月间,我们分别对中国科学院动物研究所生产的促卵泡素（每瓶10mg,售价350元）和从加拿大进口的促卵泡素（每瓶400mg,售价800元）进行超排效果对比试验。就其超排效果进行比较,以寻求较为适用的FSH制品。     

Superovulation in cycling mares using equine follicle stimulating hormone (eFSH)

Superovulation would potentially increase the efficiency and decrease the cost of embryo transfer by increasing embryo collection rates. Other potential clinical applications include improving pregnancy rates from frozen semen, treatment of subfertility in stallions and mares, and induction of ovulation in transitional mares. The objective of this study was to evaluate the efficacy of purified equine follicle stimulating hormone (eFSH; Bioniche Animal Health USA, Inc., Athens, GA) in inducing superovulation in cycling mares. In the first experiment, 49 normal, cycling mares were used in a study at Colorado State University. Mares were assigned to 1 of 3 groups: group 1, controls (n = 29) and groups 2 and 3, eFSH-treated (n = 10/group). Treated mares were administered 25 mg of eFSH twice daily beginning 5 or 6 days after ovulation (group 2). Mares received 250 (of cloprostenol on the second day of eFSH treatment. Administration of eFSH continued until the majority of follicles reached a diameter of 35 mm, at which time a deslorelin implant was administered. Group 3 mares (n = 10) received 12 mg of eFSH twice daily starting on day 5 or 6. The treatment regimen was identical to that of group 2. Mares in all 3 groups were bred with semen from 1 of 4 stallions. Pregnancy status was determined at 14 to 16 days after ovulation.In experiment 2, 16 light-horse mares were used during the physiologic breeding season in Brazil. On the first cycle, mares served as controls, and on the second cycle, mares were administered 12 mg of eFSH twice daily until a majority of follicles were 35 mm in diameter, at which time human chorionic gonadotropin (hCG) was administered. Mares were inseminated on both cycles, and embryo collection attempts were performed 7 or 8 days after ovulation.Mares treated with 25 mg of eFSH developed a greater number of follicles (35 mm) and ovulated a greater number of follicles than control mares. However, the number of pregnancies obtained per mare was not different between control mares and those receiving 25 mg of eFSH twice daily. Mares treated with 12 mg of eFSH and administered either hCG or deslorelin also developed more follicles than untreated controls. Mares receiving eFSH followed by hCG ovulated a greater number of follicles than control mares, whereas the number of ovulations from mares receiving eFSH followed by deslorelin was similar to that of control mares. Pregnancy rate for mares induced to ovulate with hCG was higher than that of control mares, whereas the pregnancy rate for eFSH-treated mares induced to ovulate with deslorelin did not differ from that of the controls. Overall, 80% of mares administered eFSH had multiple ovulations compared with 10.3% of the control mares.In experiment 2, the number of large follicles was greater in the eFSH-treated cycle than the previous untreated cycle. In addition, the number of ovulations during the cycle in which mares were treated with eFSH was greater (3.6) than for the control cycle (1.0). The average number of embryos recovered per mare for the eFSH cycle (1.9 ± 0.3) was greater than the embryo recovery rate for the control cycle (0.5 ± 0.3).In summary, the highest ovulation and the highest pregnancy and embryo recovery rates were obtained after administration of 12 mg of eFSH twice daily followed by 2500 IU of hCG. Superovulation with eFSH increased pregnancy rate and embryo recovery rate and, thus, the efficiency of the embryo transfer program.

Introduction

Induction of multiple ovulations or superovulation has been an elusive goal in the mare. Superovulation would potentially increase the efficiency and decrease the cost of embryo transfer by increasing embryo collection rates.[1 and 2] Superovulation also has been suggested as a critical requirement for other types of assisted reproductive technology in the horse, including oocyte transfer and gamete intrafallopian transfer. [2 and 3] Unfortunately, techniques used successfully to superovulate ruminants, such as administration of porcine follicle stimulating hormone and equine chorionic gonadotropin have little effect in the mare. [4 and 5]The most consistent therapy used to induce multiple ovulations in mares has been administration of purified equine pituitary gonadotropins. Equine pituitary extract (EPE) is a purified gonadotropin preparation containing approximately 6% to 10% LH and 2% to 4% FSH.[6] EPE has been used for many years to induce multiple ovulations in mares [7, 8 and 9] and increase the embryo recovery rate from embryo transfer donor mares. [10] Recently, a highly purified equine FSH product has become available commercially.The objectives of this study were to evaluate the efficacy of purified eFSH in inducing superovulation in cycling mares and to determine the relationship between ovulation rate and pregnancy rate or embryo collection rate in superovulated mares.

Materials and methods

Experiment 1

Forty-nine normally cycling mares, ranging in age from 3 to 12 years, were used in a study at Colorado State University. Group 1 (control) mares (n = 29) were examined daily when in estrus by transrectal ultrasonography. Mares were administered an implant containing 2.1 mg deslorelin (Ovuplant, Ft. Dodge Animal Health, Ft. Dodge, IA) subcutaneously in the vulva when a follicle 35 mm in diameter was detected. Mares were bred with frozen semen (800 million spermatozoa; minimum of 30% progressive motility) from 1 of 4 stallions 33 and 48 hours after deslorelin administration. The deslorelin implants were removed after detection of ovulation.[11] Pregnancy status was determined at 14 and 16 days after ovulation.Group 2 mares (n = 10) were administered 25 mg of eFSH (Bioniche Animal Health USA, Inc., Athens, GA) intramuscularly twice daily beginning 5 or 6 days after ovulation was detected. Mares received 250 g cloprostenol (Estrumate, Schering-Plough Animal Health, Omaha, NE) intramuscularly on the second day of eFSH treatment. Administration of eFSH continued until a majority of follicles reached a diameter of 35 mm, at which time a deslorelin implant was administered. Mares were subsequently bred with the same frozen semen used for control mares, and pregnancy examinations were performed as described above.Group 3 mares (n = 10) received 12 mg of eFSH twice daily starting 5 or 6 days after ovulation and were administered 250 μg cloprostenol on the second day of treatment. Mares were randomly selected to receive either a deslorelin implant (n = 5) or 2500 IU of human chorionic gonadotropin (hCG) intravenously (n = 5) to induce ovulation when a majority of follicles reached a diameter of 35 mm. Mares were bred with frozen semen and examined for pregnancy as described above.

Experiment 2

Sixteen cycling light-horse mares were used during the physiologic breeding season in Brazil. Reproductive activity was monitored by transrectal palpation and ultrasonography every 3 days during diestrus and daily during estrus. On the first cycle, mares were administered 2500 IU hCG intravenously once a follicle 35 mm was detected. Mares were subsequently inseminated with pooled fresh semen from 2 stallions (1 billion motile sperm) daily until ovulation was detected. An embryo collection procedure was performed 7 days after ovulation. Mares were subsequently administered cloprostenol, and eFSH treatment was initiated. Mares received 12 mg eFSH twice daily until a majority of follicles were 35 mm in diameter, at which time hCG was administered. Mares were inseminated and embryo collection attempts were performed as described previously.

Statistical analysis

In experiment 1, 1-way analysis of variance with F protected LSD was used to analyze quantitative data. Pregnancies per ovulation were analyzed by x² analysis. In experiment 2, number of large follicles, ovulation rate, and embryo recovery rate were compared by Student,'s t-test. Data are presented as the mean S.E.M. Differences were considered to be statistically significant at p < .05, unless otherwise indicated.

Results

In experiment 1, mares treated with 25 mg eFSH twice daily developed a greater number of follicles 35 mm in diameter (p = .001) and ovulated a greater number of follicles (p = .003) than control mares (Table 1). However, the number of pregnancies obtained per mare was not significantly different between the control group and the group receiving 25 mg eFSH (p = .9518). Mares treated with 12 mg eFSH and administered either hCG or deslorelin to induce ovulation also developed more follicles 35 mm (p = .0016 and .0003, respectively) than untreated controls. Mares receiving eFSH followed by hCG ovulated a greater number of follicles (p = .003) than control mares, whereas the number of ovulations for mares receiving eFSH followed by deslorelin was similar to that of control mares (p = .3463). Pregnancy rate for mares induced to ovulate with hCG was higher (p = .0119) than that of control mares, whereas the pregnancy rate for eFSH-treated mares induced to ovulate with deslorelin did not differ from that of controls (p = .692). Pregnancy rate per ovulation was not significantly different between control mares (54.5%) and mares treated with eFSH followed by hCG (52.9%). The lowest pregnancy rate per ovulation was for mares stimulated with 25 mg eFSH and induced to ovulate with deslorelin. The mean number of days mares were treated with 25 mg or 12 mg of eFSH was 7.8 ± 0.4 and 7.5 ± 0.5 days, respectively. Overall, 80.0% of mares administered eFSH had multiple ovulations compared with 10.3% of control mares.     

Secretion of luteinizing hormone and follicle stimulating hormone in intact and ovariectomized mares in summer and winter

Sequential samples of blood were drawn via jugular catheters every 15 min for 24 h from four mares in each of five reproductive states: intact anestrous mares in winter, intact diestrous mares in summer, intact estrous mares in summer, ovariectomized mares in winter and ovariectomized mares in summer. Estrous mares were sampled on d 4 or 5 of estrus and diestrous mares on d 10 or 11 of diestrus. Each sample of plasma was assessed for concentrations of luteinizing hormone (LH) and follicle stimulating hormone (FSH) in two independent radioimmunoassays. A computer program was developed that determined peak hormone concentrations based on assay sensitivity, assay variability and repeatability of peaks in both independent assays. Peaks in LH and FSH were observed for mares in all five reproductive states, except for FSH concentrations in estrous mares. High frequency peaks of short duration were observed only in ovariectomized mares. Low frequency peaks of relatively long duration were observed in both intact and ovariectomized mares in both seasons. With the exception of estrous mares, there was variation among mares in the patterns of LH and(or) FSH within any one group; all estrous mares exhibited high, variable LH concentrations and low, constant FSH concentrations. In general, peaks in both gonadotropins occurred simultaneously. Ovariectomized mares exhibited more (P less than .05) peaks/24 h than intact mares for both gonadotropins. Ovariectomized mares also exhibited more (P less than .05) FSH peaks/24 h in summer than in winter.     

Ovarian inhibition of peripheral plasma concentration of follicle stimulating hormone in prepuberal Holstein heifers

The objective of this study was to determine effects of age and castration on follicle stimulating hormone (FSH) secretion in prepuberal heifers. In experiment 1, twelve heifers were bilaterally ovariectomized at 3, 6, or 9 months of age (n = 4/group). Blood was collected at 10 min intervals for 8 hr at 1 week before ovariectomy and 1 and 4 weeks after ovariectomy. Frequency, amplitude and duration of FSH pulses were calculated. Mean plasma concentration of FSH (ng/ml), and frequency (pulses/8 hr), amplitude (ng/ml), and duration (min/pulse) of FSH pulses were not altered by age. Mean concentration of FSH increased (P less than .01) from 1 week before to 1 week and 4 weeks after ovariectomy, respectively, in all age groups. Pulse frequency increased (P less than .05) from 1 week before ovariectomy to 4 weeks after ovariectomy in 3 month old heifers, from 1 week before to 4 weeks after ovariectomy in 6 month old heifers, and from 1 week before to 1 week and 4 weeks after ovariectomy in 9 month old heifers. In experiment 2, twelve heifers were bilaterally ovariectomized at 3, 6 or 9 weeks of age (n = 4/group). Sample collection and measurement of mean concentration of FSH were the same as in experiment 1. Mean concentration of FSH increased (P less than .01) from 1 week before to 1 and 4 weeks after ovariectomy in heifers ovariectomized at 6 and 9 weeks of age.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of dosage and frequency of injection of luteinizing hormone releasing hormone on release of luteinizing hormone and follicle stimulating hormone in estradiol-treated steers

The objective was to determine how estradiol (0 vs 1 mg) and changes in the dosage of luteinizing hormone releasing hormone (LHRH; 1,000 ng/steer vs 1 ng/kg body weight) and frequency of LHRH injection (25 vs 50 min) affect LH and follicle stimulating hormone (FSH) release in steers. In steers pretreated with estradiol peak concentrations of LH in serum after LHRH averaged 14.4 ng/ml, which was greater (P less than .001) than peak concentrations in steers given oil (7.4 ng/ml). Increasing the dosage of LHRH from 1 ng/Kg body weight (approximately or equal to 300 ng/steer) to 1,000 ng/steer increased (P less than .001) peak LH values from 7.5 to 14.4 ng/ml. Furthermore, increasing the frequency of LHRH injections from once every 50 min to once every 25 min increased (P less than .001) LH release, but only in steers given estradiol. Estradiol reduced basal concentrations of FSH by 65% and then increased LHRH-induced FSH release by 276% (P approximately .07) relative to values for steers given oil. Only when 1,000 ng LHRH was given every 25 min to steers pretreated with estradiol were LH and FSH release profiles similar to the preovulatory gonadotropin surges of cows in magnitude, duration and general shape. The results demonstrate that increases in the dosage or frequency of LHRH pulses increase LHRH-induced release of LH, but not of FSH. Furthermore, these results are consistent with the hypothesis that in cows, estradiol increases responsiveness of the gonadotrophs to LHRH and then increases the magnitude and frequency of pulses of LHRH secretion beyond basal levels, thereby causing the preovulatory gonadotropin surges.     

Luteinizing hormone, follicle stimulating hormone and prolactin secretion in ewes and wethers after zeranol or estradiol injection

Plasma concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH) and prolactin (PRL) were determined over a 24-h period using radioimmunoassay in sheep injected with corn oil (control) or various doses of zeranol or estradiol-17 beta. Injection of .333, 1 or 10 mg of zeranol caused dose-related increases (P less than .01) in plasma PRL (peak levels at 12 to 18 h) and LH (peak levels at 12 to 20 h) in ovariectomized ewes. Similarly, PRL and LH increased following doses of 33 or 100 microgram of estradiol. Before the LH surge, plasma LH levels were significantly depressed (4 to 8 h). Plasma FSH levels were significantly decreased 4 to 8 h after zeranol and estradiol injection. Slight surges of FSH were observed at times similar to those of LH, but the peak level was never greater than control levels. Injection of 1 mg of zeranol or 100 microgram of estradiol into wethers resulted in a 24-h pattern of PRL secretion not significantly different of LH concentration and significantly prolonged inhibition of FSH secretion. These results indicate similarities in the effects of zeranol and estradiol on anterior pituitary hormone secretion within groups of animals of the same sex or reproductive state. Differences in secretion and plasma concentrations of LH, FSH and PRL due to underlying sexual dimorphism are maintained and expressed even when animals are challenged with structurally different compounds of varying estrogenic potencies.     

Effect of administration of adrenocorticotropic hormone on plasma concentrations of testosterone, luteinizing hormone, follicle stimulating hormone and cortisol in stallions

In boars and rabbits, administration of adrenocorticotropic hormone (ACTH) results in a testis-dependent, short-term increase in concentrations of testosterone in peripheral plasma. This experiment was designed to assess the short-term effects of a single ACTH injection on plasma concentrations of testosterone, luteinizing hormone (LH), follicle stimulating hormone (FSH) and cortisol in stallions. Eight light horse and two pony stallions were paired by age and weight and then one of each pair was randomly assigned to the treatment (ACTH, .2 IU/kg of body weight) or control (vehicle) group. Injection of ACTH increased (P<.01) plasma concentrations of cortisol by approximately twofold in the first 60 minutes; control stallions showed no change (P>.10) in concentrations of cortisol during the blood sampling period. Control stallions exhibited a midday increase (P>.05) in concentrations of testosterone similar to that reported previously; ACTH treatment prevented or delayed this increase such that concentrations of testosterone in treated stallions were lower (P<.05) than in controls 4 to 5 hours after injection of ACTH. Treatment with ACTH had no effect (P<.10) on plasma concentrations of LH or FSH up to 12 hours after injection.     

Concentrations of luteinizing hormone, follicle stimulating hormone and prolactin following transection of the pituitary stalk in ovariectomized ewes

Two experiments (Spring and Fall) were conducted in ovariectomized ewes to determine changes in pituitary hormone secretion immediately after pituitary stalk-transection. Ewes underwent either pituitary stalk-transection (SS), sham-transection (SH) or administration of anesthesia only (AO). Stalk-transected, but not sham-operated or anesthetized ewes had polyuria and polydipsia for 7 to 14 days after surgery. Concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH) and prolactin were measured in peripheral blood samples collected every 10 minutes for a six-hour period. Results were comparable for each season. During the six hours following surgery or removal from anesthesia, concentrations of LH declined in all ewes, but more slowly in SS ewes. No differences in patterns or mean concentrations of FSH were observed. Immediately after surgery, concentrations of prolactin were elevated, then declined in SH and SS ewes. The decrease was greater in SH than SS ewes. Data are consonant with the view that hypothalamic inhibition as well as LHRH stimulation regulate gonadotropin release by the pituitary.     

Patterns of secretion of luteinizing hormone, follicle stimulating hormone and testosterone in stallions during the summer and winter

Samples of jugular blood were drawn from each of five stallions every 15 min for 12 h during the summer and winter to determine the short-term fluctuations in plasma concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH) and testosterone. Concentrations of LH and FSH were generally not pulsatile, although one stallion exhibited three distinct pulses in these hormones during the winter. In general, patterns of secretion of all three hormones were similar in both seasons and the number of significant rises in hormonal concentrations did not differ between seasons. Concentrations of LH and FSH were positively correlated (P less than .05) for eight of the ten sampling periods, indicating a close relationship between the secretion rates of these two gonadotropins. Testosterone concentrations varied in an episodic manner during the 12-h period, and all stallions exhibited at least one episode of high testosterone secretion regardless of the pattern of LH concentrations. The response in testosterone concentrations to the three LH pulses exhibited by the one stallion in winter was not the same for each pulse. The correlations between a single random sample and mean concentrations over the 12-h period were high (r between .88 and .99) for all three hormones, indicating that a single sample of blood would be representative of overall concentrations. It appears that the stallion differs from males of other domestic species in that concentrations of gonadotropins and testosterone vary in a much less pulsatile manner.     

Effects of butorphanol on luteinizing hormone and follicle stimulating hormone levels in ovariectomized rats--comparative study with morphine sulphate.

Studies were conducted into the effects on pituitary gonadotrophic hormones in ovariectomized rats of butorphanol, a synthetic morphine derivative which was claimed to be a potent analgesic with few side-effects, in comparison to effects of the naturally occurring alkaloid morphine. For this purpose, 3 groups of ovariectomized rats were used. Rats of the 1st group were injected butorphanol at 2 dose levels (1 or 2 mg/kg body weight [b.w.]. Those of the 2nd group were injected morphine sulphate (10 or 20 mg/kg b.w.). The 3rd group was injected saline and served as control. Blood samples were collected by orbital sinus punctures, just before treatment and 1 hour post injection. Luteinizing hormone (LH) and follicle-stimulation hormone levels were determined in the sera of rats by radio-immuno-assay. The results revealed that morphine, at the 2 dose levels used, produced more than 90% decrease in serum LH concentration, whereas butorphanol produced more than 70% decrease in serum LH levels. Both morphine and butorphanol, at the 2 doses used, produced more than 76% decrease in serum follicle stimulating hormone concentration. It is concluded that butorphanol, the morphinic derivative, has a depressive effect on the synthesis and/or release of gonadotrophic hormones. This inhibitory effect, however, was nearly as potent as that produced by morphine sulphate.     

Testosterone propionate treatment of stallions: Effects on secretion of luteinizing hormone and follicle stimulating hormone in daily samples and after administration of gonadotropin releasing hormone

Ten stallions were used to determine if the stallion responds to administration of testosterone propionate (TP) with an increase in follicle stimulating hormone (FSH) secretion after administration of gonadotropin releasing hormone (GnRH) as has been previously observed for geldings and intact and ovariectomized mares. Five stallions were treated with TP (350 μg/kg of body weight) in safflower oil every other day for 11 days; control stallions received injections of safflower oil. The response to GnRH (1.0 μg/kg of body weight) was determined for all stallions before the onset of treatment (GnRH I) and at the end of treatment (GnRH II). Blood samples were also withdrawn daily from 3 days prior to treatment through GnRH II. Treatment with TP decreased (P<.10) concentrations of FSH in daily blood samples. However, treatment with TP did not affect (P>.10) the GnRH-induced secretion of FSH. Concentrations of luteinizing hormone (LH) decreased (P<.05) in daily blood samples averaged over both groups of stallions and were lower (P<.10) in TP-treated stallions than in controls during the latter days of treatment. We conclude that TP administration to stallions does not alter the FSH response to GnRH as has been observed for geldings and for mares of several reproductive states.     

Effects of adding luteinizing hormone to a medium containing follicle stimulating hormone on progesterone-induced differentiation of cumulus cells during meiotic resumption of porcine oocytes

In the present study, we investigated the effects of adding luteinizing hormone (LH) to a medium containing follicle stimulating hormone (FSH) on the shift in expression of progesterone receptor (PR) isoforms (PR‐A and PR‐B) and the roles in function of cumulus cells of cumulus‐oocyte complexes (COC). The level of PR‐B mRNA in cumulus cells was up‐regulated by FSH during the first 16‐h cultivation but the level was significantly decreased at 20 h. The decrease of PR‐B mRNA was accelerated when COC were cultured with FSH and LH. Still, a high level of total PR mRNA was maintained in cumulus cells cultured with or without the addition of LH up to 20 h, suggesting that the expression of PR isoforms was shifted from PR‐B to PR‐A in cumulus cells. The reduction of PR‐B was also induced by addition of progesterone to FSH‐containing medium. The addition of LH or progesterone to FSH‐containing medium stimulated cumulus expansion of COC as compared with that of COC cultured with FSH. In the expanded COC, ADAMTS‐1 which is expressed in granulosa cells and cumulus cells in rodent follicles through LH‐induced progesterone‐ and PR‐dependent pathway, was more accumulated within the COC matrix. These results suggest that the addition of LH or progesterone to FSH‐containing medium is required for the differentiation of cumulus cells, such as cumulus expansion, mediated by the shift from PR‐B to PR‐A in them.     

Pubertal development in the male pig: effects of treatment with a long-acting gonadotropin-releasing hormone agonist on plasma luteinizing hormone, follicle stimulating hormone and testosterone.

The effects of a long-acting gonadotropin-releasing hormone (GnRH) agonist, [D-Trp6]-GnRH (GnRH-A) on developmental profiles of plasma luteinizing hormone (LH), follicle stimulation hormone (FSH) and testosterone (T), and pituitary responsiveness to exogenous GnRH were studied in male Dutch Landrace x Large White crossbred pigs from 1 to 30 wk of age. Group 1 control animals (control; n = 12) were injected subcutaneously in the neck with vehicle at 1 and 16 wk of age. Group 2 animals (early treatment; n = 10) were injected with 600 micrograms [D-Trp6]-GnRH at 1 wk and with vehicle at 16 wk. Group 3 animals (late treatment; n = 8) were injected with vehicle and 3 mg GnRH-A at 1 and 16 wk, respectively. Group 4 animals (early plus late treatment; n = 9) were injected at both 1 and 16 wk with GnRH-A. Blood was collected by brachiocephalic puncture at weekly or biweekly intervals, and through brachiocephalic cannulae, to determine longitudinal profiles of LH, FSH and T, and plasma gonadotropin responses to intravenous injection of GnRH (0.1 microgram/kg), respectively. In control animals, LH and FSH declined over the first 5 wk of postnatal life and peaked again at 10-14 wk. Levels of both hormones were basal from 18 to 30 wk. Plasma T was high in the first week, declined progressively over the next few weeks and remained low until 24 wk when a transient increment was noted. The LH and FSH responses to acute GnRH stimulation were similar at 7 and 14 wk and declined significantly at 23 wk of age.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparative efficacy of an injectable vaccine and an intranasal vaccine in stimulating Bordetella bronchiseptica-reactive antibody responses in seropositive dogs

OBJECTIVE: To compare antibody responses to intranasal and SC Bordetella bronchiseptica vaccines in seropositive dogs. DESIGN: Randomized controlled study. ANIMALS: 40 young adult Beagles vaccinated against B bronchiseptica. PROCEDURE: Dogs were randomly assigned to 1 of 4 groups (intranasal vaccine, SC vaccine, intranasal and SC vaccines, no vaccine) and vaccinated on day 0. Serum and salivary B bronchiseptica-reactive antibody responses were measured on days 0 through 7, 10, 14, 21, and 28. RESULTS: Dogs that were vaccinated with the SC vaccine, alone or in combination with the intranasal vaccine, had a significant increase in serum concentration of B bronchiseptica-reactive IgG beginning on day 5 and persisting through day 28. Dogs that were vaccinated with the intranasal vaccine alone had a significant increase in serum concentration of B bronchiseptica-reactive IgG beginning on day 10 and persisting through day 28, but serum IgG concentration in these dogs was significantly less than concentration in dogs that received the SC vaccine. Neither vaccine had a demonstrable effect on salivary concentrations of B bronchiseptica-reactive IgA or IgG. On day 10, all vaccinated groups had significantly higher serum IgA concentrations than did unvaccinated control dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the SC B bronchiseptica vaccine may be used to stimulate antibody responses in seropositive dogs. There was no apparent benefit to administering these vaccines simultaneously. Intranasal vaccines may not be effective for booster vaccination of dogs previously exposed to or immunized against B bronchiseptica. Dogs should be vaccinated at least 5 days prior to exposure to B bronchiseptica.