野生鲇鱼生长激素分泌的季节变化及其神经内分泌调控

采用离体垂体碎片灌流孵育系统 ,将处于性腺退化期野生鲇鱼垂体切成约 1mm3 的碎片 ,用M 199冲洗之后放入灌流柱的两层Cytodex -Ⅲ微载体之间 (温度为 19± 1℃ )。每 5分钟收集一管灌流液 ,- 2 5℃贮存待测GH。采用鲤鱼GH放射免疫测定方法 (cGHRIA)测定鲇鱼垂体碎片灌流液以及血清和垂体中的GH含量。结果表明 :促黄体素释放激素类似物 [desGly10 (D Ala6)LHRHethylamide ,LHRH A]不能显著刺激离体垂体碎片基础GH分泌 ,注射LHRH A后不能显著提高血清基础GH水平 ;注射DA能显著提高鲇鱼血清基础GH水平 ,APO能以剂量依赖方式显著刺激垂体碎片基础GH分泌。雌、雄鲇鱼血清GH水平在 6月达到峰值 ,垂体GH水平在 3月和 7月份各出现一个峰值 ,各个季节雌鱼垂体和血清GH水平均显著高于雄鱼。鲇鱼血清和垂体GH水平与生殖周期有密切联系。     

鳗鲡繁殖生物学研究Ⅳ．人工催熟过程中下海鳗鲡的GtH分泌活动、性腺发育状况和脑垂体GtH细胞的超显微结构

雌二醇通过正反馈作用能促进脑垂体促性腺激素(GtH)细胞的合成活动,使脑垂体GtH水平显著升高。促黄体素释放激素的类似物(LHRH-A)和利血平(reserpine,RES)能促进脑垂体GtH细胞的分泌活动,使血液中GtH含量显著升高。鲤垂体、人体绒毛膜促性腺激素(HCG)和LHRH—A三种激素混合进行多次注射能诱导雌雄鳗鲡性腺发育成熟,其催熟效果明显优于它们的分别单独多次注射或者鲤鱼脑垂体和HCG的多次注射,表明外源的和内源的促性腺激素对于诱导鳗鲡性腺发育成熟都是重要的。鳗鲡脑垂体GtH细胞超显微结构的观察证实它们在激素诱导性腺发育成熟过程中处于活跃的合成与分泌状态。     

多巴胺能药物对虎纹蛙GnRH和LH分泌活动的影响

利用在体注射实验和放射免疫测定法,研究了多巴胺能药物对性腺处于再发育期虎纹蛙的促性腺激素释放激素（GnRH)及促黄体激素（LH)分泌活动的影响。结果是：多巴胺（DA）及其激素剂阿扑吗啡（APO）可显著降低血浆LH水平;而多巴胺的拮抗剂－地欧酮（DOM）可显著增加垂体LH含量。DA对脑中cGnRH-Ⅱ的合成有抑制作用,而OM对其mGnRH的释放有一定的刺激作用。结果表明：DA可在脑及垂体水平分别抑制虎纹蛙GnRH和LH的释放,DA对LH释放的抑制作用很可能是通过D2受体实现的。     

高效能的促黄体素释放激素类似物对金鱼促性腺激素分泌的作用

金鱼对IRH-A反应有明显的季节性变化,在早春(2月)生殖季节开始前诱导的血清GtH含量最高;在生殖季节过后(5-6月)GtH的释放反应减弱,而在夏季(8月)性腺处于退化状况时,反应很弱或者没有反应。LRH-A注射剂量的高低对血清GtH含量增高的影响并不都很明显,但高剂量(1.0微克／克体重)能诱导较高的血清GtH含量并能维持24小时。经过多次注射高剂量以后,脑垂体GtH含量下降。温度、注射剂量、注射间隔时间都和诱导的血清GtH含量有密切关系。连续9-10天每日注射LRH-A,能刺激性腺退化的金鱼恢复性腺发育,但对性腺已开始发育的金鱼,却抑制或减慢其性腺进一步发育。     

高效能的促黄体素释放激素类似物对金鱼促性腺激素分泌的作用

金鱼对LRH-A反应有明显的季节性变化,在早春(2月)生殖季节开始前诱导的血清GtH含量最高;在生殖季节过后(5—6月)GtH的释放反应减弱,而在夏季(8月)性腺处于退化状况时,反应很弱或者没有反应。LRH-A注射剂量的高低对血清GtH含量增高的影响并不都很明显,但高剂量(1.0微克/克体重)能诱导较高的血清GtH含量并能维持24小时。经过多次注射高剂量以后,脑垂体GtH含量下降。温度、注射剂量、注射间隔时间都和诱导的血清GtH含量有密切关系。连续9—10天每日注射LRH-A,能刺激性腺退化的金鱼恢复性腺发育,但对性腺已开始发育的金鱼,却抑制或减慢其性腺进一步发育。       

鲑鱼促性腺激素释放激素类似物和Domperidone诱导几种养殖鱼GtH分泌和排卵的研究

鲑鱼GnRH的一种类似物:(D-Arg~6,Trp~7,Leu~8,Pro~9NET)-LHRH(sGnRH—A)和多巴胺拮抗物domperidone对诱导鱼类GtH分泌和排卵具有非常高的活性。sGnRH-A和DOM同时一次注射能促使大鳞副泥鳅和鲤鱼的GtH大量迅速释放,并有效地诱导它们以及草鱼、鲢鱼、鳙鱼、鲮鱼等排卵或产卵。     

多巴胺拮抗物PIM增强丘脑下部促黄体素释放激素类似物诱导大鳞副泥鳅排卵效应的研究

在注射低剂量LRH—A的同时注射多巴胺拮抗物pimozide(PIM),能显著增强LRH-A诱导的排卵效应,排卵率达到100％。LRH—A PIM能促进脑垂体GtH细胞的合成和分泌,而注射多巴胺能抑制LRH—A诱导的GtH释放活动。     

睾酮对日本鳗鲡离体下丘脑-脑垂体复合物GtH合成与分泌的影响

用离体孵育的方法研究了睾酮（T）对日本鳗鲡（Anguilla japonica)完整的正丘脑-脑垂体复合物（hypothalamus-pituiary complex,HPC)、分离的下丘脑（hypothalamus,H)加脑垂体（pituitary,P)以及单独的脑垂体（pituitary,P)促性腺激素（GtH)合成与释放的影响,在不加T的孵育液中孵育,HPC组的孵育液及其脑垂体中的GtH含量最高,H P组次之,而P组最低,表明下丘脑通过GnRH直接刺激脑垂体GtH的合成与释放,在加入T的孵育液中孵育,P组的孵育液及脑垂体中的GtH含量显著增加,并且和孵育液中的T呈剂量依存的正反馈作用,当T和H与P一起孵育时,低浓度的T（0.1和1μmol/L)刺激HPC组和H P组的GtH释放,呈现正反馈作用,而高浓度的T（10μmol/L)则抑制HPC组和H P组的GtH释放,表现负反馈作用。这些结果直接证明日本鳗鲡下丘脑对脑垂体GtH分泌的调控以及性类固醇激素（如雄鳗的睾酮）对脑垂体GtH分泌的反馈作用。     

长臀鮠生长激素的生殖周期变化

国内外学者对硬骨鱼类的GH的季节变动及神经内分泌调控有一定的研究,其中在鲤科和鲑鳟鱼类研究的较为深入,近来一些学者对鲇形目的胡子鲇科、科和鲇科做了一定的研究,研究结果表明,GH分泌和神经内分泌调节机理与鲤科鱼类在某些方面存在一定差异。如鲤科鱼类中,GnRH起着GH释放因子的作用,但非洲鲇鱼和鲇鱼中却未证实GnRH刺激GH释放,并且鲇鱼中GH分泌的季节变化与生殖之间有密切联系。       

帕金森病模型大鼠脑内多巴胺与铁含量的关系

实验采用原子吸收分光光度法,快速周期伏安法,高效液相电化学检测等方法,研究以6－羟基多巴（6－OHDA）制备的帕金森病（PD）模型大鼠黑质内铁含量的变化。铁对多巴胺（DA）能神经元的直接毒性作用以及铁离子螯合剂甲磺酸去铁胺的神经保护作用。结果发现：（1）PD大鼠损毁侧黑质内铁含量为非标准PD大鼠的3倍左右;（2）PD大鼠损毁侧纹状体内铁含量无明显改变;（3）单纯注射6－OHDA的大鼠其损毁侧纹状体（CPu)DA的释放量和含量均明显降低;（4）侧脑室预先注射甲磺酸去铁胺,再重复上述实验,损毁侧CPu DA释放量和含量均无明显改变;（5）单侧黑质内注射40ug FeCl3后,大鼠损毁侧CPu内DA释放量和含量显著降低。上述结果提示,6－OHDA可导致CPu DA释放量及含量减少,此过程有铁的参与。由于铁可导致DA神经元死亡,因此铁含量的增加可能是DA含量减少的原因之一,甲磺酸去铁胺具有保护DA神经元的作用。     

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.     

多巴胺能药物对鲤鱼促性腺激素分泌活动的影响

研究多巴胺（ＤＡ）及其激动剂阿扑吗啡（ＡＰＯ）和拮抗剂Ｄｏｍｐｅｒｉｄｏｎｅ（ＤＯＭ）对不同性腺发育时期和不同年龄雌鲤促性腺激素（ＧｔＨ）分泌活动的影响。结果表明：鲤鱼基础ＧｔＨ分泌具有明显的季节性变化,表现为血清ＧｔＨ含量在性腺发育早期较低,性腺成熟期升高,到性腺退化期又降低。ＤＡ和ＡＰＯ对ＧｔＨ分泌的抑制作用及ＤＯＭ对ＧｔＨ分泌的刺激作用在不同性腺发育时期为：性腺发育早期〉性腺成熟期〉性腺退化     

诱导大鳍鱯和长吻鮠排卵过程中血清GTH水平的变化

繁殖期从嘉陵江收集性成熟的大鳍■ 和长吻 野生亲鱼,用Ｌｉｎｐｅ方法（即ＬＨＲＨ－Ａ加多巴胺Ｄ２受体拮抗剂地欧酮）或传统的ＬＨＲＨ—Ａ加脑垂体的方法进行催产,定时取血样,用放射免疫方法测定催产过程中血清ＧＴＨ水平的变化,进一步证实鲇形目鱼类ＧＴＨ的分泌受到下丘脑分泌的促性腺激素释放激素ＧｎＲＨ和多巴胺的双重调节;排卵和产卵也是以血清ＧＴＨ的急剧升高为先导的,而最终能否排卵还有赖于血清ＧＴＨ峰是否超过“排卵阈值”。尽管催产后的大鳍 和长吻 雄鱼血清ＧＴＨ水平也有一个高峰出现,但血清ＧＴＨ水平升高幅度都大大低于雌鱼,这种现象在硬骨鱼类可能具有普遍性。     

注射促黄体素释放激素类似物和地欧酮诱导大鳍■和长吻■排卵的研究

对于性成熟的大鳍Ｍｙｓｔｕｓｍａｃｒｏｐｔｅｒｕｓ（Ｂｌｅｅｋｅｒ）野生鱼,单独注射多巴胺的抑制剂地欧酮（ＤＯＭ）不能影响血清促性腺激素（ＧＴＨ）水平,也不能诱导排卵;单独注射类似物ＬＨＲＨ－Ａ,虽能使血清ＧＴＨ水平显著升高,但仅产生较低的排卵率;而当ＤＯＭ与ＬＨＲＨ－Ａ结合注射却显著增强ＬＨＲＨ－Ａ促进血清ＧＴＨ水平升高的作用,并诱导出较高的排卵率。对性成熟的长吻ＬｅｉｏｃａｓｉｓｌｏｎｇｉｒｏｓｔｒｉｓＧｕｎｔｈｅｒ野生鱼,使用ＬＨＲＨ－Ａ＋ＤＯＭ作２次注射诱导排卵的效果也与注射ＬＨＲＨ－Ａ加脑垂体这一传统诱导排卵方法相似。Ｌｉｎｐｅ方法（ＬＨＲＨ－Ａ＋ＤＯＭ,作１次或２次注射）避免了采集、保存脑垂体不便给生产带来的麻烦,在科鱼类的人工繁殖中,具有较高的推广价值。     

多巴胺拮抗物PIM增强丘脑下部促黄体素释放激素类似物诱导大鳞副泥鳅排卵效应的研究

在注射低剂量LRH-A的同时注射多巴胺拮抗物pimozide(PIM),能显著增强LRH-A诱导的排卵效应,排卵率达到100％。 LRH-A+PIM能促进脑垂体GtH细胞的合成和分泌,而注射多巴胺能抑制LRH-A诱导的GtH释放活动。     

Catecholamines and pituitary function. VII: Effects of acute and chronic dopamine-receptor blockade on pituitary response to TRH-GNRH in normal women and in patients with hyperprolactinemic amenorrhea

To investigate whether an enhanced dopamine (DA) inhibition on pituitary thyrotrophs and gonadotrophs may account for the abnormal TSH and LH dynamics in pathological hyperprolactinemia, we examined the effect of an acute lysis of the putative DA overinhibition, as obtained with continuous domperidone (DOM) infusion, on both basal and TRH-GnRH stimulated PRL, TSH and LH release in both normal cycling women and patients with pathological hyperprolactinemia. The effect of TRH-GnRH administration was also examined in women with DA-antagonist induced hyperprolactinemia, in order to evaluate the effect of a chronic lack of the physiological DA inhibition on pituitary hormone dynamics. Patients with both pathological and DA-antagonist induced hyperprolactinemia displayed an evident TSH and LH hyper-responsiveness to TRH-GnRH. The PRL response was reduced in the former but enhanced in the latter group. Domperidone infusion resulted in a marked increase in serum PRL levels in normal cycling women, but not in patients with pathological hyperprolactinemia. The abolition of the putative DA-overinhibition at the pituitary level with DOM infusion in patients with pathological hyperprolactinemia was followed by a slight increase in basal TSH output but did not modify the TSH and LH hyperresponsiveness to TRH-GnRH. The similarities in TSH and LH dynamics between patients with pathological and DA-antagonist induced hyperprolactinemia and the ineffectiveness of DOM infusion in modifying the TSH and LH hyper-responses to TRH-GnRH in the former group, seem to exclude the widely accepted idea that endogenous DA overactivity is responsible for the abnormal thyrotroph and lactotroph dynamics in women with hyperprolactinemic amenorrhea.     

Calcium ions as a mediator in GnRH action on gonadotropin release in the common carp (Cyprinus carpio L)

Collagenase-dispersed carp pituitary cells in a perifusion system were used to study the role of calcium ions in the mechanism of GnRH action on the release of maturational gonadotropin (GtH) in fish. The specific calcium chelator EGTA and the calcium antagonist manganese (Mn2+) caused a 40% inhibition in the basal GtH release and completely blocked GnRH-stimulated GtH release. Short-term application of graded doses of calcium ionophore A23187 caused a dose-dependent increase in GtH secretion. A23187 failed to stimulate GtH secretion in the presence of EGTA. Depolarization of the membrane by K+ caused a strong stimulation of GtH release similar to the action of GnRH. Stimulatory action of K+ was inhibited by EGTA. These data suggest a role for extracellular calcium as an intracellular mediator in GnRH-stimulated, as well as in basal, GtH release in carp. The stimulation of GtH release by K+ also indicates that voltage-dependent processes could be involved in this phenomenon.     

促性腺激素释放激素及多巴胺对斜带石斑鱼生长激素分泌及其mRNA表达的调控

研究斜带石斑鱼生长激素分泌及其mRNA表达的调控规律对于性别分化的控制、临床药物的选择,以及石斑鱼的增养殖等均具有重要的理论意义和实践意义。本文应用静态孵育系统,采用放射免疫测定法和化学发光液相杂交实验,研究GnRH和DA对斜带石斑鱼GH分泌、GHmRNA合成的调控作用。100nmol／LsGnRH作用斜带石斑鱼脑垂体碎片1也4h,明显促进GH的释放和GHmRNA的合成,并具有时间依存性;10nmol／L～1μmol／LsGnRH作用1h能明显促进斜带石斑鱼脑垂体释放GH,促进GHmRNA的合成,表现出明显的剂量效应。100nmol／L、1μmol／LmGnRH作用1h以一定的剂量依存方式促进GH的释放、促进GHmRNA的合成,但mGnRH的效应比相应剂量的sGnRH的作用弱。APO为DA受体的非选择性激动剂,不同剂量APO对斜带石斑鱼脑垂体碎片的作用结果显示,10nmol／L-1μmol／L APO以剂量依存方式促进斜带石斑鱼脑垂体碎片释放GH、促进GHmRNA的合成：1μmol／LAPO作用12h以上明显促进GH的释放和GHmRNA的合成,并随时间的延长而增加。与sGnRH对斜带石斑鱼GH释放、GHmRNA合成的作用相比,APO的作用较弱。本文研究结果证实GnRH和DA能促进斜带石斑鱼脑垂体GH释放和GHmRNA合成。     

A presumptive role for GABA in the stimulatory effects of Des-Gly10, [D-Ala6]-LHRH-ethylamide and pimozide on the gonadotropin release in carp

To investigate the effect of endogenous gamma-aminobutyric acid (GABA) on the blood maturating gonadotropin (GtH) levels, or to study its interaction with pimozide (dopamine antagonist) and a luteinizing hormone-releasing hormone analog (LHRH-a), sexually mature male and female carps were treated with drugs that may either inhibit GABA biosynthesis or GABA degradation. In females the irreversible inhibitor of GABA-transaminase, gamma-vinyl GABA (GVG), which was to increase the endogenous GABA-ergic tone, had no influence on GtH release. On the other hand, the increased GtH response to the combination of pimozide (PIM) and LHRH-a was clearly enhanced by the administration of 3-mercaptopropionic acid (MPA), an inhibitor of the rate limiting enzyme of GABA-biosynthesis. In males the GABA-ergic compound, valproic acid (DPA) decreased LHRH-a stimulated GtH levels. In male carps that received PIM to diminish the dopaminergic inhibition of GtH release, the spermiating response to LHRH-a was increased by administration of MPA. These data suggest that GABA interacts with the action of dopamine and the gonadotropin releasing hormone (GnRH) on the release of GtH.     

Pure uptake blockers of dopamine can reduce prolactin secretion: studies with diclofensine

The effects of diclofensine, a pure dopamine (DA) uptake inhibitor on 1) 3H-DA uptake in rat arcuate-periventricular nucleus-median eminence synaptosomes, 2) basal and K+-evoked endogenous DA release from tuberoinfundibular dopaminergic (TIDA) neurons and 3) in vivo prolactin (PRL) secretion were studied. Diclofensine, in concentrations of 0.01, 0.1 and 1 microM caused a marked decrease of 3H-DA uptake. In addition, it was unable to stimulate basal endogenous DA release which, on the contrary, was elicited by d-amphetamine in the same concentration (50 microM). On the other hand, diclofensine (50 microM) caused a 3 fold enhancement of K+-evoked DA release. Finally, the compound, when administered in vivo to male rats, significantly reduced basal serum PRL levels. The results of the present study seem to indicate that the pharmacological blockade of DA uptake in TIDA neurons is a condition sufficient to cause a reduction of PRL release.