Amino Acid Neurotransmitters and Dopamine in Brain and Pituitary of the Goldfish: Involvement in the Regulation of Gonadotropin Secretion

An isocratic high-performance liquid chromatographic technique was developed to measure levels of gamma-aminobutyric acid (GABA), glutamate, and taurine in the brain and pituitary of goldfish. Accuracy of this procedure for quantification of these compounds was established by evaluating anesthetic and postmortem effects and by selectively manipulating GABA concentrations by intraperitoneal administration of the glutamic acid decarboxylase (GAD) inhibitor 3-mercaptopropionic acid or the GABA transaminase inhibitor gamma-vinyl GABA. The technique provided a simple, rapid, and reliable method for evaluating the concentrations of these amino acids without the use of complex gradient chromatographic systems. To investigate the relationship between neurotransmitter amino acids and the control of pituitary secretion of gonadotropin, the effects of injection of taurine, GABA, or monosodium glutamate on GABA, glutamate, taurine, and, in some instances, monoamine concentrations in the brain and pituitary were evaluated and related to serum gonadotropin levels. Injection of taurine caused an elevation in serum gonadotropin concentrations. In addition, injection of the taurine precursor hypotaurine but not the taurine catabolite isethionic acid elevated serum gonadotropin levels. Intracerebroventricular injection of either GABA or taurine also elevated serum gonadotropin concentrations. Pretreatment of recrudescent fish with alpha-methyl-p-tyrosine reduced pituitary dopamine concentrations and also potentiated the serum gonadotropin response to taurine. Injection of monosodium glutamate caused an increase of glutamate content in the pituitary at 24 h; this was followed by a decrease at 72 h after administration. Pituitary GABA, taurine, and dopamine concentrations underwent a transient depletion after monosodium glutamate administration, and this was associated with an elevation of serum gonadotropin content.(ABSTRACT TRUNCATED AT 250 WORDS)     

Brain lesions and short-term endocrine effects of monosodium L-glutamate in goldfish,Carassius auratus

Summary Monosodium L-glutamate (MSG) was injected intraperitoneally into goldfish at a dosage of 2.5mg/g body weight. At 24 h post-injection there was a marked hypertrophy and edema in the region of the nucleus lateralis tuberis (NLT) from the anterior margin of the pituitary stalk through to the posterior end of the NLT, irrespective of the sex of the goldfish. A similar hypertrophy and edema occurred ventral to the anterior commissure in the preoptic region in the anterior-ventral nucleus preopticus periventricularis (NPP). At 6 h post-injection a slight vacuolization was evident in these two regions, and at two days the hypertrophy and edema had abated from the extent observed at 24 h post-injection. At five and eight days post-injection only necrotic cells were found in the affected NLT region, but only a small band of necrotic cells was evident in the anterior-ventral preoptic region. No other brain lesions were evident. Serum levels of gonadotropin (GtH) were increased at 6 h, 24 h, and two days after treatment with MSG, but were similar to control values at five, seven and eight days after MSG in male and female goldfish. Exocytosis of small dark secretory granules in gonadotrophs was evident at 24 h after MSG in a fish with a somewhat greater increase in serum GtH than usually found. The time course of increased serum GtH levels postinjection of MSG is consistent with the observed time course of hypertrophy and atrophy of NLT neurons; the increase in serum levels of GtH is interpreted to reflect a stimulation of release of GtH-releasing factor from neurons in the NLT. Electron microscope investigation indicates that prolactin cells have increased secretory and synthetic activity from 24 h through to seven days post-injection of MSG. The mechanism for stimulation of the prolactin cells by MSG is not known. No other changes in activity of adenohypophysial secretory cells were found.Supported by grant A-6371 from the Natural Sciences and Engineering Research Council of CanadaCNRS France and NRC Canada Exchange Scientist.     

长臀（鱼危）脑垂体和血清中促性腺激素的生殖周期变化

鲇形目鱼类在世界养殖鱼类中占有重要的位置.     

促性腺激素释放激素类似物对长臀鮠脑垂体促性腺激素分泌的影响

采用离体灌流孵育技术和促性腺激素的放射免疫测定方法,对长臀鮠(Cranoglanis bouderius)脑垂体碎片促性腺激素的分泌进行了研究。结果表明：持续的促性腺激素释放激素类似物(GnRH-A)能显著刺激退化期的长臀鮠离体脑垂体碎片促性腺激素(GTH)的分泌,并且长臀鮠脑垂体碎片对持续的GnRH-A刺激未表现出脱敏性,该结果与胡子鲇和鲇鱼相似,而与金鱼和鲤科鱼类不同;重复脉冲GnRH-A刺激对长臀鮠脑垂体碎片GTH分泌具有促进作用,而且存在剂量依存关系,与鲇鱼和鲤科鱼类相类似。上述结果表明在长臀鮠的人工繁殖中可以用持续高浓度GnRH-A刺激对长臀鮠进行催熟和催产。     

Plasma levels of gonadotropin releasing hormone during menstrual cycle ofMacaca radiata

A sensitive radioimmunoassay for gonadotropin releasing hormone has been developed. The assay has been validated for its specificity by testing various analogues of gonadotropin releasing hormone. Analysis of plasma samples during the menstrual cycle of 4 female bonnet monkeys showed a significant increase in the immunoreactive gonadotropin releasing hormone levels during preovulatory period of the menstrual cycle.     

不同的下丘脑肽和神经递质对鲤鱼促性腺激素和生长激素分泌活动的影响

以１龄性腺发育中期鲤鱼为材料,采用腹腔（ｉ．ｐ）注射的方法,研究不同的下丘脑肽和神经递质对鲤鱼促性腺激素（ＧｔＨ）和生长激素（ＧＨ）分泌的影响。结果表明：促甲状腺激素释放激素（ＴＲＨ）、Ｌ－多巴（Ｌ－ＤＯＰＡ）、甲基睾酮（ＭＴ）、γ－氨基丁酸（ＧＡＢＡ）、促黄体素释放激素类似物（ＬＨＲＨ－Ａ）和三碘甲状腺原氨酸（Ｔ３）都能显著刺激ＧｔＨ的分泌,但最大效应时间各不相同。ＴＲＨ和ＬＨＲＨ－Ａ能促进ＧＨ的分泌,Ｌ－ＤＯＰＡ、ＭＴ、ＧＡＢＡ对血清ＧＨ水平没有明显影响;Ｔ３则对ＧＨ分泌有一定的抑制作用。这说明鲤鱼ＧｔＨ和ＧＨ的分泌除了受各自的下丘脑释放因子和释放抑制因子的双重神经内分泌调控外,还受多种其它相同和不同调节因子的影响,也反映了鲤鱼ＧｔＨ和ＧＨ分泌的神经内分泌调控的复杂性。     

Purification of equine gonadotropins and comparative study of their acid-dissociation and receptor-binding specificity

A method for the simultaneous purification of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) from equine pituitaries is briefly described. Different forms of each hormone were obtained. The total yield of LH was 24.2 mg·kg^?1 with a recovery of 22% and the yield of FSH was 26 mg·kg^?1 with a recovery of 34%. The specific activities of both hormones, measured in homologous equine radio-receptor assays are equal to or higher than those of the preparations described so far. In all species studied so far the acid-dissociation curves of LH and FSH are similar; this is an agreement with the view that the binding of the common α-subunit and the specific β-subunits involves polypeptide regions which are identical in both hormones. In contrast, the acid-dissociation pK_a of equine LH was found to be considerably lower (3.9) than that of equine FSH (5.8). The equine gonadotropins exhibit a much lower specificity with receptors of a porcine testicular fraction compared with an equine fraction. Equine LH exhibited a binding activity on FSH receptors from a porcine testicular fraction equal to 20% that of equine FSH instead of only 1% for an equine binding fraction. Similarly, all the equine FSH preparations tested exhibited a five-fold higher binding-activity on porcine LH receptors than on equine LH receptors. In the porcine system, pregnant mare serum gonadotropin behaved like equine LH towards LH and FSH receptors. In contrast, on equine binding fraction, pregnant mare serum gonadotropin was only 4% as active as equine LH and was devoid of FSH activity. All the data we have obtained are consistent with the ‘negative specificity’ model we proposed recently.     

Relationships between follicular fluid steroid hormone concentrations, oocyte maturity, in vitro fertilization and embryonic development in the rhesus monkey

Oocytes and matched samples of follicular fluid (FF) were obtained from 70 follicles of five rhesus monkeys stimulated with either pregnant mare serum gonadotropin or human menopausal gonadotropin. Follicular aspiration was performed 30-32 h after human chorionic gonadotropin administration. The concentrations of estradiol (E2), progesterone (P), testosterone (T), and dihydrotestosterone (DHT) in FF were measured. Twenty-six percent of oocytes were classified as mature (M), 41% matured in vitro (Miv), 13% were dysmature, and 20% atretic. M oocytes were associated with significantly higher levels of P and a higher P:E2 ratio. There were no differences in hormone levels associated with fertilized and nonfertilized oocytes. Thirty-five embryos developed to the six- to eight-cell stage in vitro, of which 13 exhibited optimal cleavage rates. Significantly lower levels of E2 and higher P:E2 ratios were associated with the more rapidly cleaving embryos. Proportionally more embryos showing optimal cleavage rates developed from M compared to Miv oocytes, and only embryos derived from M oocytes developed to blastocysts in culture. Optimal cleavage rates to the six- to eight-cell stage in vitro, rather than fertilization rates, are a better indicator of (subsequent) developmental capacity, and, in this study, embryonic development was closely associated with the maturity of the oocyte at recovery.     

Regulation of ovarian 17β-hydroxysteroid dehydrogenase activity by gonadotropin

Serum testosterone levels are elevated prior to the lutropin surge, and decline abruptly following the release of endogenous lutropin. To investigate this phenomenon, the activity of 17β-hydroxysteroid dehydrogenase, the enzyme directly related to testosterone production form androstenedione, was measured. This was done in immature rats in which follicular maturation and ovulation were induced by pregnant mare serum gonadotropin administration. It appears that the effect of the gonadotropin on the enzyme activity is sharply divided into two phases that match with the follicular and the luteal phases. One day following gonadotropin administration, there was already a 7.67-fold increase in the original activity which further increased 48 h following hormone administration. At the peak of the lutropin surge, when follicular development is at its maximum, a 18.44-fold increase was measured. The activity fell abruptly 10 h following ovulation, at a time when fresh corpora lutea are already present in the ovary.It seems that the evaluation of serum testosterone followed by its abrupt decline, is directly related to the increased and decreased ovarian 17β-hydroxysteroid dehydrogenase activity. The possible importance of the observed changes to the mechanisms of the onset of puberty are discussed.     

脊椎动物促性腺激素释放激素的分子结构变异型和功能多样性

综述了近十几年来有关对脊椎动物促性腺激素释放激素的结构和功能的研究进展,阐明促性腺激素释放激素分子结构的变异型和它们在脊椎动物的分布和进化,分析促性腺激素释放激素的多种功能及其受体的结构与特异性。     

Ovarian follicular differentiation with prepubertal gonadotropin surges and gonadotropin priming in mice

Preantral follicles were mechanically isolated from the ovaries of 1.5 to 8 week old mice and cultured in vitro for 4 days in the presence or absence of either activin A or FSH. Plasma gonadotropin, estradiol and immunoreactive (IR) inhibin levels were measured. Cultured follicles showed stepwise changes in response to recombinant human (rh) FSH, with no response until 11 days, a gradual increase from 2 weeks, culminating in a strong response to rhFSH at 8 weeks. The response to activin A was vice versa. It enhanced the effect of rhFSH on preantral follicular growth of up to 4-week-old mice, but inhibited the effect of rhFSH in 8-week-old mice. The peak of the prepubertal gonadotropin surge was observed on day 11. Seven-day-old mice were treated with either luteinizing hormone releasing hormone (LHRH) or rhFSH or human chorionic gonadotropin (hCG) for 3 consecutive days from day 7, and follicles were collected on day 11. Those follicles showed enhanced response to rhFSH, no response to activin A, and an enhanced response to the combination of rhFSH and activin A, suggesting that the chronological changes in follicular response are a result of the prepubertal gonadotropin surge.     

溴隐亭不同给药方案治疗高泌乳素血症女性性腺激素促排卵临床研究

目的:研究溴隐亭不同给药方案在治疗高泌乳素血症(HPRL)女性不育症中的临床疗效,关注其对女性促性腺激素诱导排卵的影响。方法:本研究共纳入60例就诊于我院的确诊为高泌乳素血症不孕不育患者,随机分为两组。分为研究组与对照组:研究组采取先口服溴隐亭调整血清泌乳素水平至正常后予以促性腺激素诱导排卵;对照组采取促性腺激素与溴隐亭同步治疗方案。结果:观察两组患者的促排卵周期数、平均用药天数、雌二醇水平及妊娠率,两组治疗前后的血清泌乳素都显著改善(P0.05);但是两组之间相比,采取溴隐亭药物治疗后诱导排卵的研究组在促排卵、雌二醇水平和妊娠率方面具有显著优势(P0.05)。结论:采用溴隐亭治疗高泌乳素血症患者,调整至正常后再使用促卵泡激素药物促排卵治疗不孕不育具为较优的治疗方案。     

Cd^2＋对离体的鲤鱼脑垂体分泌促性腺激素的影响

采用离体的鲤鱼脑垂体灌流孵育系统,研究重金属镉离子对脑垂体分泌促性腺激素的害影响。发现Ｃｄ＾２＋既能直接刺激ＧｔＨ分泌,又抑制鲑鱼促性腺激素类似物释放激素刺激脑垂体分泌ＧｔＨ作用。实验结果提示,Ｃｄ＾２＋对鲤鱼脑垂体ＧｔＨ分泌细胞的毒定机制之一可能是它干扰了Ｃａ＾２＋的正常生理作用。     

Peripheral administration of metastin induces marked gonadotropin release and ovulation in the rat

Metastin is a novel peptide that has been isolated from the human placenta as the cognate ligand of the G-protein-coupled receptor OT7T175 (or GPR54). However, its physiological functions have not yet been fully investigated. In the present study, we show that subcutaneous administration of metastin increased the plasma levels of gonadotropins (follicle-stimulating hormone and luteinizing hormone) and induced ovulation in prepubertal female rats that had been pretreated with pregnant mare serum gonadotropin to induce follicle maturation. Furthermore, metastin administration drastically increased the plasma levels of gonadotropins in male rats. This action was abolished by pretreatment with a GnRH antagonist, and was accompanied by induction of c-Fos immunoreactivity in GnRH neurons. These results suggest that s.c. administered metastin induces the release of gonadotropin via activation of the hypothalamic GnRH neurons.     

Hormonal induction of ovulation stimulates atresia of antral follicles in a vespertilionid bat, Scotophilus heathi

Scotophilus heathi is a seasonally monoestrous subtropical vespertilionid bat found at Varanasi, India. Although the antral follicles remain present in the ovaries of S. heathi from November till March, ovulation is delayed in this species until early March. In order to understand the mechanism of ovulation suppression during this period of delayed ovulation, the effects of human chorionic gonadotropin (hCG), pregnant mare's serum gonadotropin (PMSG), follicle stimulating hormone (FSH) and gonadotropin releasing hormone agonist (GnRH agonist) on ovarian morphology and steroid concentration were investigated. Hormonal treatments were given as a single i.p. dose 24 h after capture. The bats were sacrificed 48 h after the injection. Treatment with hCG, PMSG, FSH and GnRH agonist failed to induce ovulation in S. heathi, although these hormones produced a high degree of ovarian stimulation. The administration of hCG and PMSG induced ovarian enlargement, intense hyperemia, marked changes in the interstitial cells (ICs), development of several antral follicles and a varying degree of abnormalities in the oocytes of most of the antral follicles. In the bats treated with hCG, PMSG and GnRH agonist, androstenedione concentration increased significantly to extraordinarily high levels, whereas estradiol concentration decreased. Administration of FSH caused regression of ICs and pyknosis of granulosa cells in the majority of antral follicles. FSH did not enhance androstenedione concentration. The results of the present study suggest that the failure of hormonal treatments to induce ovulation during the period of delayed ovulation might be due to a seasonal desensitization of ovarian follicles in S. heathi. The hormonal treatment instead stimulated the ICs to produce a high level of androstenedione resulting in atretic changes of the antral follicles.     

Fine structure and function of isolated gonadotropic cells as revealed from pituitaries of immature rainbow trout,Salmo gairdneri,by means of a new enzymatic dispersion technique

A procedure has been developed for dissociating pituitary glands of juvenile rainbow trout, Salmo gairdneri, producing a preparation of single dispersed pituitary cells in which morphological and functional integrity is preserved. The pituitaries are dispersed by sequential treatment with 0.1% collagenase, 0.04% ethylene-diamine-tetra-acetic acid (EDTA) and 0.125% dispase. The cell yield is 0.3--0.35 x 10(6) cells per pituitary with a cell viability percentage of 95 +/- 1% and single cell percentage of 87 +/- 4%. The isolated cells are kept in a suspension system and the gonadotropic cells are identified by the double antibody immuno-enzyme-cytochemical technique using anti-carp-beta-gonadotropin as first antibody. Secretory activity is estimated by measuring the gonadotropin content in cells and culture media by radioimmunoassay. Isolated cells show an autonomy of gonadotropin secretion. 17 alpha-Methyltestosterone both in vivo and in vitro stimulates the production of gonadotropin in the cells and seems to inhibit its release from the cells. It is concluded that this in vitro system can be used as a model for studying the control of gonadotropic cells in juvenile rainbow trout.     

The insulin-like growth factor (IGF) system and gonadotropin regulation: actions and interactions

Insulin-like growth factors (IGF) are polypeptides that regulate growth, differentiation and survival in a multitude of cells and tissues. The IGF system consists of ligands, receptors, binding proteins and binding protein proteases. The influence of the IGF system on reproductive parameters, specifically gonadotropin release and interactions between the IGF system and other effectors of gonadotropin release will be examined in this review.     

腹腔镜联合促性腺激素释放激素激动剂治疗子宫内膜异位症伴不孕症的疗效观察

目的:探讨腹腔镜联合促性腺激素释放激素激动剂治疗子宫内膜异位症伴不孕症的疗效。方法:将我院2013年2月~2014年2月收治的200例子宫内膜异位症伴不孕症患者随机分为2组各100例,对照组采用曲普瑞林治疗,观察组采用腹腔镜联合曲普瑞林治疗,检测患者治疗前后血清雌二醇(E2)、癌胚抗原125(CA125)及基质金属蛋白酶-1(MMP-1)水平,对比两组治疗后疼痛视觉评分(VAS)、盆腔包块和用药停止后月经恢复情况、随访1年记录两组复发率及妊娠率。结果:治疗后观察组VAS评分、盆腔包块和月经恢复情况均明显优于对照组,差异有统计学意义(P0.05);两组患者血清CA125、E2及MMP-1水平均显著下降,且观察组下降更明显,差异有统计学意义(P0.05);随访1年后,观察组妊娠率明显高于对照组,复发率明显低于对照组,差异有统计学意义(P0.05)。结论:腹腔镜联合促性腺激素释放激素激动剂治疗子宫内膜异位症的疗效显著,能有效降低复发率、提高妊娠率,值得临床推广应用。