Pulsatile administration of gonadotropin releasing hormone (Gn-RH) to lactating sows using a portable automatic pump ("Zyklomat")

The object of this investigation was to study the feasibility of using a “Zyklomat roller peristaltic pump” for pulsatile administration of gonadotropin releasing hormone (Gn-RH) in primiparous lactating sows. Four primiparous sows were used. The pump catheter was inserted into a jugular vein on day 8 or day 22 of lactation. The pump delivered a 1 min Gn-RH pulse of 10 μg every 89 min for 7 days. The pump worked without any complication or detrimental effect on the sows throughout the experimental period. Two sows showed standing oestrus during lactation and one of them ovulated. It can be concluded that the “Zyklomat” pump can be used for pulsatile infusion of Gn-RH in lactating sows.     

Response of the lactating and postweaning sow to gonadotropin releasing hormone (GnRH)

Clinical and endocrinological responses to administration of gonadotropin releasing hormone analog (LH-RH-A) during the lactation period and postweaning in the sow were investigated. Plasma LH concentrations in lactating sows rose immediately after administration of LH-RH-A. However, in postweaning sows the increase of LH level was more slowly. Three of 5 postweaning sows came into estrus and ovulated after LH-RH-A treatment. One sow exhibited a distinct LH response, but her ovaries remained quiescent. The remaining one with feeble estrus for a short period became cystic ovaries. Thus, LH response to GnRH in the sow seems to be higher during early lactation than at 2 days postweaning.     

Ovarian response to pregnant mare serum gonadotropin and porcine pituitary extract in gilts actively immunized against gonadotropin releasing hormone

Two experiments were conducted to determine the effect of exogenous gonadotropins on follicular development in gilts actively immunized against gonadotropin releasing hormone (GnRH). Four gilts, which had become acyclic after immunization against GnRH, and four control gilts were given 1,000 IU pregnant mare serum gonadotropin (PMSG), while four additional control gilts were given saline. Control animals were prepuberal crossbred gilts averaging 100 kg body weight. Control gilts given saline had ovaries containing antral follicles (4 to 6 mm in diameter). Control gilts given PMSG exhibited estrus and their ovaries contained corpora hemorrhagica and corpora lutea. PMSG failed to stimulate follicular growth in gilts immunized against GnRH, and ovaries contained regressed corpora albicantia and small antral follicles (less than 1 mm in diameter). Concentrations of luteinizing hormone (LH) and estradiol-17 beta (E2) were non-detectable in gilts immunized against GnRH and given PMSG. In the second experiment, five gilts actively immunized against GnRH were given increasing doses of PMSG every third day until unilateral ovariectomy on d 50. PMSG failed to stimulate follicular growth, and concentrations of follicle stimulating hormone (FSH), E2 and LH were not detectable. Six weeks later, gilts were given a booster immunization and then were given 112 micrograms LH and 15 micrograms FSH intravenously every 6 h for 9 d. The remaining ovary was removed on d 10. Although LH and FSH concentrations were elevated, administration of gonadotropins did not stimulate follicular growth or increase E2 concentrations. These results indicate that neither PMSG or exogenous LH and FSH can induce E2 synthesis or sustain follicular development in gilts actively immunized against GnRH.     

Gonadotropin releasing hormone (GnRH) induced luteinizing hormone (LH) secretion from perifused equine pituitaries.

In vitro responsiveness of the horse anterior pituitary (AP) gonadotropes to single and multiple GnRH challenges was examined. The pituitaries were collected from reproductively sound mares in estrus (n = 5) and diestrus (n = 5). Uniform 0.5 mm AP slices were subdivided using a 3 mm biopsy punch and then bisected for use in the perifusion chamber. Four bisected sections per chamber were perifused at 0.5 ml/min at 37 C for 560 min in Medium 199 saturated with 95% 0(2)/5% CO2. Ten minute fractions were collected after an initial 2 hr equilibration period. Four different treatment regimes of GnRH (10(-10) M) were evaluated: (A) three consecutive 10 min GnRH pulses separated by 80 and 100 min, respectively; (B) a single 120 min GnRH infusion; (C) a 10 min GnRH pulse followed 80 min later by a 120 min GnRH infusion and (D) two 10 min GnRH pulses separated by 60 min followed 80 min later by a 120 min GnRH infusion. Estimated total pituitary LH content was higher in estrous than diestrus mares (p less than 0.05). The total amount of LH released in response to GnRH tended to be greater in estrus than diestrus (p less than 0.1), whereas the percentage of LH released in estrus and diestrus was similar. An increase in the area under the LH response curve was noted with each successive 10 min pulse of GnRH during both estrus and diestrus (p less than 0.05), demonstrating a self-priming effect of GnRH. In addition, a significant increase in the peak LH amplitude (p less than 0.05) and the slope to peak amplitude (p less than 0.05) were observed for the 120 min GnRH pulse in regime C and D indicating that prior exposure to short-term pulses of GnRH increased the acute LH secretory response. These results suggest that in the cycling mare (1) the responsiveness of the pituitary (amount of LH released as percent of total LH) is similar in both estrus and diestrus, however, the magnitude of the LH response (total microgram amount of LH released) differs with the stage of the estrous cycle, being highest in estrus, and appears to be related, in part, to pituitary LH content and (2) GnRH self-priming occurs independently of the stage of the estrous cycle. Furthermore, we have demonstrated that the pulsatile mode of GnRH can act directly on the anterior pituitary to dictate the pulsatile release pattern of LH in the cycling mare.     

The influence of exogenous pituitary growth hormone on the ovulatory response to PMSG and hCG and the LH response to estradiol benzoate in prepubertal gilts

Two experiments were performed to examine the influence of exogenous growth hormone on the reproductive axis in gilts. Experiment one employed 26 Yorkshire × Landrace prepubertal gilts, which were selected at 150 d and 86.5 ± 1.5 kg bodyweight (BW) and assigned equally to two treatments. Gilts received injections of either porcine growth hormone at 90 μg/kg BW, or vehicle buffer, from 150 to 159 d. At 154 d gilts received 500 IU PMSG, followed 96 hr later by 250 IU hCG. Gilts were slaughtered at 163 days and their ovaries recovered to determine ovulatory status. In each treatment, gilts failed to show any ovarian response to PMSG/hCG. All remaining control gilts ovulated and their ovaries appeared morphologically normal. In gilts receiving exogenous growth hormone, fewer ovaries (4/11, P<.01) appeared morphologically normal. The ovaries of all other growth hormone injected gilts had very large (12–25 mm) non-luteinized follicles. In experiment two, 20 prepubertal Yorkshire × Landrace gilts were selected at 138 days and 85 kg BW. These gilts received injections of growth hormone at 90 μg/kg BW (n=9) or vehicle (n=11) from 138 to 147 days. At 143 days, all gilts were given an injection of estradiol benzoate (EB) at 15 μg/kg BW. Blood samples were taken at the time of EB injection, at 24 and 36 hr and then at 6 hr intervals until 78 hr. All samples were assayed for serum LH concentrations. The EB induced LH peak height was lower (P<.04) in gilts receiving exogenous growth hormone than in controls. The results presented indicate that the daily injection of growth hormone at 90 μg/kg BW reduced the estradiol-induced release of LH in addition to reducing the number of corpora lutea in gonadotrophin stimulated gilts.     

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.     

Stimulation of annexin A5 expression by gonadotropin releasing hormone (GnRH) in the Leydig cells of rats

The distribution and regulation of annexin A5 expression, a gonadotropin releasing hormone (GnRH) receptor regulated protein in gonadotropes and luteal cells, in the testes of rats were examined. Immunocytochemical staining revealed high levels of annexin A5 in the Leydig and endothelial cells and lower levels in the primary spermatocytes and sperm. Hemicastration significantly increased the annexin A5 content of the remaining testis within 24 h. Annexin A5 immunoreactivity was increased mainly in interstitial tissues including the peritubular cells, while some spermatocytes also showed higher intensity of annexin A5 in the remaining testis. Administration of hCG (50 IU) enhanced the testicular content of annexin A5 after 24 h. This treatment expanded the area of interstitial tissue in the testis and increased annexin A5 immunoreactivity, but the area of the of endothelial cells was unchanged. Similarly, human chorionic gonadotropin (hCG) enhanced annexin A5 expression in a primary culture of testis cells that consisted of mainly interstitial cells. Because GnRH stimulates the expression of annexin A5 in the gonadotropes and luteal cells, we examined the effect of GnRH on annexin A5 expression in the testes. We found that des-Gly10 [Pro9]-GnRH ethylamide (100 nM), a GnRH agonist, increased annexin A5 expression in cultured testis cells and that Cetrorelix (100 nM), a GnRH antagonist, inhibited the effect of hCG on annexin A5 expression. These results suggest that pituitary luteinizing hormone promotes annexin A5 synthesis in Leydig cells and that this effect could be mediated by local GnRH in the testis.     

Seasonal differences in the parameters of luteinizing hormone release to exogenous gonadotropin releasing hormone in prepubertal Holstein heifers in Sapporo

Stress due to summer heat has adverse effects on reproduction in Holstein dairy cattle. Summer suppression of reproduction of Holsteins can pose an important economic problem, even in Hokkaido prefecture located in the northern region of Japan. Hokkaido is one of the most important dairy farming areas of Japan. This study is an attempt to clarify the seasonal differences in the parameters of luteinizing hormone (LH) response to exogenous gonadotropin releasing hormone (GnRH) in Sapporo, Hokkaido, Japan. A total of 12 prepubertal heifers received an injection with GnRH analogue intramuscularly in either May (n=4, May group), July (n=4, July group), or November (n=4, November group), and serial blood samples were collected to analyze the parameters of the LH response curve after GnRH injection. The parameters were as follows: the basal LH concentration, peak LH concentration, duration from the time of GnRH injection to the time of the peak LH concentration, and area under the LH response curve (AUC). There were no significant differences in the basal and peak LH concentrations or the AUC among the three groups. The July group reached the LH peak significantly (P<0.05) faster than the May group, but there was no significant difference with the November group. Therefore, the results of the present study do not demonstrate an effect of summer heat on the LH response to the exogenous GnRH in Holstein heifers.     

Luteinizing hormone in the bovine pars tuberalis: secretion in response to luteinizing hormone releasing hormone and intracellular isoforms

The objective of this study was to examine the physiological characteristics of gonadotropes in the bovine (b) pars tuberalis as assessed by their ability to release Luteinizing Hormone (LH) in response to LH-Releasing Hormone (LHRH) and the intracellular distribution of LH isoforms. At slaughter, the stalk median eminence and associated pars tuberalis as well as the anterior pituitary gland were collected from each of 7 castrate males. Each stalk median eminence and pituitary gland was mid-sagitally sectioned and weighed. One half of each tissue was immediately frozen and subsequently homogenized to determine the intracellular distribution of bLH isoforms. Tissue extracts were desalted by flow dialysis against water and chromatofocused on pH 10.5-7.0 gradients. The remaining half of the pituitary was sliced with a Staddie-Riggs slicer. The pituitary slices and the remaining half of the stalk median eminence were perifused (0.1 ml/min) for a total of 360 min with effluent samples (1.0 ml) collected every 10 min. At 130 min tissues were stimulated with 5 x 10(-8) M LHRH. Concentrations of LH in the effluent samples and the fractions collected from chromatofocusing were determined by radioimmunoassay. The release of LH in response to LHRH was 43.9% and 47.0% above basal secretion for the pars tuberalis and pituitary, respectively, suggesting similar degrees of responsiveness. Pars tuberalis and pituitary extracts resolved into nine LH isoforms during chromatofocusing and were coded with letters beginning with the most basic form. No differences (P greater than .05) were observed in distribution of LH isoforms between the pars tuberalis and the pituitary gland.(ABSTRACT TRUNCATED AT 250 WORDS)     

Plasma luteinizing hormone response to increasing doses of synthetic gonadotrophin-releasing hormone in heifers

The dosenresponse relationship for a synthetic gonadotrophin-releasing hormone (GnRH) was studied in normally cycling heifers using the area under the luteinizing hormone (LH) curve as a response parameter. Oestrus was synchronized by an injection of 0.5 mg cloprostenol before the experiment started and after the 3rd treatment with GnRH. Treatment with GnRH as assigned in a Latin square included 5 dose levels (0, 10, 50, 100, 250 μg) and 5 treatment days over a period of 22 days. GnRH was capable of inducing an increase of plasma LH within 30 min after injection. Plasma LH response increased with increasing doses of GnRH, the largest increase being observed when the dose was raised from 50 μg to 100 μg. One heifer did not respond to any of the doses applied. The existence of an individual treshold dose of GnRH is suggested.     

Cortisol response to two different doses of intravenous synthetic ACTH (tetracosactrin) in overweight cats

Fifteen middle-aged to older, overweight cats attending a first-opinion clinic were investigated to rule out hyperadrenocorticism as a cause of their weight problem, using two different protocols for the adrenocorticotropic hormone (ACTH) stimulation test. The cats received intravenous synthetic ACTH (tetracosactrin) at an initial dose of 125 microg; a second test was performed between two and three weeks later, using a dose of 250 microg intravenously. The mean basal serum cortisol concentration was 203 nmol/litre (range 81 to 354 nmol/litre). The highest mean serum cortisol concentration occurred at 60 minutes following the 125 microg dose and at 120 minutes following the 250 microg dose. There was, however, no statistically significant difference between these peak cortisol concentrations attained using either dose of tetracosactrin. A significantly higher mean serum cortisol concentration was attained after the higher dose at the 180 minutes time point, indicating a more prolonged response when compared with the lower dose. The cats were followed up for one year after the initial investigations and none were found to develop hyperadrenocorticism during this time.     

The effect of gonadotropin releasing hormone (GnRH) and analogs on the conception rate in cattle. A critical literature review

The objective of this article was to review the current literature of the influence of gonadotropin releasing hormone (GnRH) and GnRH agonists on conception rate in dairy cattle. The application of GnRH or agonists at artificial insemination (first and subsequent) and between days 7 and 34 after parturition were considered. The variations between studies were discussed as well as different mechanisms concerning the influence of GnRH for establishing pregnancy. From a critical point of view the routine use of GnRH or agonists at the time of first or subsequent breeding or during the postpartum period cannot be supported.     

日粮能量对初情期前母猪外周血清FSH、LH释放的影响

选杜长大三元杂交母猪24头(140日龄)分3个组,高能组、中能组、低能组。试验处理20 d,每5 d采血1次,离心,-20℃冰箱储存,然后放射免疫分析(RIA)。结果表明:外周血浆FSH、LH浓度在不同日粮能量水平饲养后,高能组急剧上升,到第10天达到高峰,然后逐渐下降,在处理第20天接近恢复试验前水平。低能组缓慢降低,第10天到最低点,然后逐渐恢复,在处理第20天接近恢复试验前水平。外周血浆FSH、LH浓度在高能组与低能组于处理第5天差异显著(P<0.05),第10天差异极显著(P<0.01),说明能量通过下丘脑-垂体-卵巢轴影响初情期前母猪生殖机能。     

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.     

In utero programming of sexually differentiated gonadotrophin releasing hormone (GnRH) secretion

It has long been recognised that steroids can have both organisational and activational effects on the reproductive neuroendocrine axis of many species, including the sheep. Specifically, if the ovine foetus is exposed to testosterone during a relatively short 'window' of in utero development (from approximately day 30-90 of a 147 day pregnancy) the neural mechanisms regulating gonadotrophin releasing hormone (GnRH) secretion become organised in a male-specific manner. In post-natal life the consequences of foetal androgen exposure are sexually differentiated responses of the GnRH neuronal network to activation by factors such as photoperiod and ovarian steroid hormones. Studies in the gonadectomized lamb have demonstrated that elevated concentrations of oestrogen (E) are unable to trigger a preovulatory-like GnRH surge in the male and the androgenized ewe lamb. Further, these animals have markedly reduced sensitivity to the inhibitory actions of progesterone on tonic GnRH release compared with normal ewes. The reasons for these abnormal steroid feedback mechanisms may reside in sexually dimorphic inputs to the GnRH neurone, including those from oestrogen-receptive neurones in the arcuate nucleus that synthetize the neuropeptide, neurokinin B (NKB). The consequences of in utero androgen exposure are reflected in a progressive and dramatic impairment of fertility in the ovary-intact ewe.