鲤鱼脑垂体中促性腺激素含量的周年变化

经丙酮干制的垂体粉用40％乙醇抽提，冷冻干燥，用放射免疫测定法测定鲤鱼垂体中促性腺激素含量的周年变化。年平均含量为56．1微克／毫克。2月份临近产卵期时，雌、雄鲤鱼垂体中的促性腺激素含量都达到最大值(雌鱼为154．0微克／毫克，雄鱼为160．0微克／毫克)。雄鱼9月份含量最低(8．3微克／毫克)，雌鱼10月份含量最低(3．8微克／毫克)，并分别在10月份和11月份后开始逐渐上升。其周年变化并与其性腺的发育成正相关关系。     

养殖银鲳性腺发育规律和性类固醇激素变化

为掌握养殖银鲳的性腺发育规律及其与性类固醇激素水平的关系,本研究采用组织切片技术和酶联免疫吸附法(Elisa),观察养殖条件下银鲳的精巢与卵巢发育特征和血清中性类固醇激素的周年变化,并分析性腺成熟指数(GSI)、肝重指数(HSI)和肥满度(CF)与性腺发育的关系。结果显示,养殖银鲳雌鱼1龄即可发育成熟,成熟卵巢呈一对""形的囊状结构。11月份有近一半个体发育至III期,33.3%已发育至IV期,12月—翌年1月越冬期间约2/3退化至II期,3月又迅速发育至IV期,4月份达到V期并产卵,5月份产卵结束卵巢进入恢复期IV期。银鲳精巢属小叶型,III期精巢出现小叶腔,IV期精巢出现精子,精巢发育速率快于卵巢,即"雄性早熟",7月龄约20%个体可达到性成熟,越冬时退化至III期,3月重新发育,精子分批成熟排精。雌鱼GSI值为0.19%~12.89%,HSI为0.97%~2.30%,CF为2.30~3.08 g/cm~3,雄鱼的GSI为0.08%~2.62%,HSI值为0.73%~1.83%,CF为2.11~2.80 g/cm~3,GSI值在V期时达到最大,HSI和CF值则于IV期达到峰值。雌鱼血清中雌二醇(E2)周年表达水平为23.27~59.13 pg/m L,雄鱼为15.90~36.20 pg/m L,雌鱼的睾酮(T)为14.57~68.67 nmol/L,雄鱼的T为18.62~66.49 nmol/L,E2水平与HSI值呈现显著正相关关系,T表达水平与GSI值呈现显著正相关关系。研究表明,银鲳血清中性类固醇激素含量与精巢和卵巢的发育密切相关,可作为了解养殖银鲳性腺发育的重要指标。     

注射含HCG和CPE的W／O／W复乳诱导日本鳗鲡性腺发育成熟的研究

用含HCG及鲤脑垂体（PG）匀浆液提取物（CPE）的W／O／W复乳，对塘养雌鳗进行注射催熟实验，雌鳗平均GSI达35．18％，最高达56．5％，其催熟效果较常规注射方法（平均GSI 27．9％，最高50．4％）好，用含LHRH－A，HCG，CPE的复乳和DOM生理盐水混悬剂对部分达到性成熟的鳗鲡进行催产，用W／O／W复乳诱导成熟的鳗鲡，排卵率为75％，用常规方法催熟的鳗鲡排卵率为55％，注射复乳制剂后，鳗鲡血清GtH含量可较长时间的保持较高水平，血清GtH的变化较常规注射方法缓和，注射乳剂还可使鳗鲡脑垂体GtH含量升高。     

镉对鲤血清促性腺激素和生长激素的影响

采用ＣｄＣｌ２浸没法处理鲤，研究镉对鲤血清促性腺激素（ＧｔＨ）和生长激素（ＧＨ）水平的影响，结果表明，较镐浓度镉（ＣｄＣｌ２９ｍｇ／Ｌ）处理引起血清ＧｔＨ水平降低，ＧＨ水平升高，提高水中钙浓度，能使由镉引起的ＧｔＨ水平的降低恢得取正常水平，并对镉引起的ＧＨ水平升高起延迟作用，镉处理使鲤对ＬＨＲＨ－Ａ刺激ＧｔＨ分泌的反应性下降，提高水中钙浓度也不能恢复正常，实验结果提示，镉引起的ＧｔＨ和ＧＨ的变化可     

血浆促性腺激素和性类固醇的年、日变化与真骨鱼类性腺周期的关系

刺激鱼类的各种外界因子综合作用于中枢神经系统。它们与下丘脑——垂体——性腺系统相互作用，因此在一定程度上，随着激素分泌的变化，促进配子形成和产卵。     

鲤脑垂体和HCG诱导雌性日本鳗鲡性腺成熟过程中血清生化成分的变化

使用鲤脑垂体和HCG对雌性日本鳗鲡人工催熟,用临床医学检验方法,测定分析了雌性日本鳗鲡卵巢发育过程中,血清总蛋白(TP)、血清甘油三脂(TG)、血清胆固醇(TC)、血糖(Glu)、血钙(Ca)和血清无机磷(P)6项生化指标的水平及变化趋势,并与对应的不同卵巢发育阶段的性腺成熟系数(GSI)进行比较分析。结果表明,84d的注射过程中,实验组的性腺逐渐成熟,GSI显著升高,第12针时达到46.32%±1.75%;而对照组性腺GSI一直在2.42%±0.18%。实验组和对照组血清TP水平相对平稳,各组变化无显著差异(P>0.05)。实验组和对照组血脂在56d前差异显著(P<0.05):实验组血清TG,TC一直维持较高水平,平均分别为(12.67±2.83)mmol/L和(15.54±3.01)mmol/L;对照组TG和TC一直下降且水平较低,平均分别为(6.33±4.14)mmol/L和(8.53±3.30)mmol/L。实验组和对照组血清Glu均在实验初的较低水平基础上持续上升。相关分析表明,实验组血清TG与GSI水平极显著负相关(r=0.732,n=6,P<0.01)。实验组血清Ca与GSI(r=0.961,n=6,P<0.01),血清P与GSI(r=0.775,n=6,P<0.01)都呈现出极显著的相关性:血清Ca和P在28d后持续上升,GSI逐渐增大。对照组中的血清Ca和P水平则一直维持在一个较低的水平。研究结果表明:雌性日本鳗鲡卵巢发育与其脂类代谢密切相关,无机离子(Ca,P)在此过程中起到重要作用。利用繁殖鱼体营养代谢特点,检测日本鳗鲡血脂或血清Ca和P水平,可成为初步判断雌性日本鳗鲡发育情况的新方法。     

17α-甲基睾酮对草鱼性腺发育及性类固醇激素水平的影响

利用养殖草鱼雌性个体所表现出的生长优势,探讨通过性逆转培育全雌草鱼品种的方法,具有较好的应用前景。实验采用拌料投喂诱导方式,在饲料中设置0、50、100和200 mg/kg 4个浓度梯度的17α-甲基睾酮(MT),连续投喂150 d,观测不同处理水平草鱼性腺结构和血液中类固醇激素雌二醇(E2)、睾酮(T)和11-酮基睾酮(11-KT)含量的变化规律,探讨草鱼性逆转过程中的特征变化。性腺的组织切片结果显示,从投喂30 d开始,各处理组卵巢中开始出现精原细胞。30～150 d,50 mg/kg MT处理组草鱼卵母细胞的生成及成熟受到抑制,发育迟缓;100 mg/kg MT处理组的生殖细胞受抑制程度要高于50 mg/kg MT处理组,实验结束时性腺已被大量精原细胞占据,有向精巢发育的趋势;200 mg/kg MT处理组精卵兼性状态较50 mg/kg MT处理组明显,但精原细胞的数量少于100 mg/kg MT处理组,对照组卵巢未出现精原细胞。血清类固醇激素测定结果显示,投喂不同剂量的MT,E_2浓度均显著升高,于45 d时达到最高值,之后趋于平稳下降,但仍显著高于对照组,未经处理的雄性草鱼,随着早期精巢的发育,其E_2水平也逐渐升高。随着MT投喂时间的延长,各处理组T浓度分别有不同程度的降低,45 d时各处理组降幅最大,且与MT浓度呈正相关,11-KT则有不同程度升高。研究表明,100 mg/kg MT为其性逆转较适宜的浓度,MT可使早期雌性草鱼血清E_2升高,发挥雄激素效应,从而促进卵巢卵细胞退化,精原细胞增殖。     

盐度驯化对雌性日本鳗鲡形态特征及性腺发育的影响

为查明盐度驯化对5龄雌性日本鳗鲡(Anguilla japonica)形态特征及性腺发育的影响,通过150 d的模拟盐度驯化实验,分析雌鳗在不同盐度驯化下的形态特征和性腺发育状况。结果显示,从盐度0～35依次驯化至90d(在盐度15、25、35依次分别驯化30d)时,雌鳗的胸围(PC)、胸鳍指数(PI)、性体指数(GSI)和肝脏指数(LI)随盐度的升高均显著增加(P<0.05);而消化道指数(DTI)和眼径指数(EI)随盐度的升高均无明显变化(P>0.05)。通过对卵巢切片观察发现,雌鳗的卵母细胞直径(OD)随盐度的升高显著上升(P<0.05),但卵母细胞始终处于脂肪泡时期。在盐度35中继续驯化,发现雌鳗在第90、120和150天的上述7项指标在各盐度组间均不存在显著性差异(P>0.05)。为了解流水对雌鳗性腺发育的影响,设置了同步盐度驯化的流水组。结果显示,在整个盐度驯化过程中,流水组雌鳗7项指标的变化趋势与静水组一致,除卵母细胞直径(OD)外的其他6项指标与同一驯化盐度下的静水组均无显著性差异(P>0.05)。流水组的卵母细胞直径(OD)均显著高于同一...     

不同激素对雌性花鳗鲡性体指数和发育过程中促性腺激素基因(GtHα,FSHβ,LHβ)表达的影响

为评估不同激素诱导花鳗鲡卵巢发育的效果并探究促性腺激素在花鳗鲡发育过程中的作用,实验比较了不同激素组合处理后花鳗鲡性体指数(gonadosomatic index,GSI)的变化,并采用实时荧光定量PCR(qRT-PCR)对处于不同发育阶段雌鱼脑垂体中促性腺激素基因GtHα(促性腺激素亚基),LHβ(促黄体生成素亚基)及FSHβ(卵泡刺激素亚基)的表达进行了定量研究.结果发现,鲤脑垂体萃取液(CPE)与人类绒毛膜促性腺激素(hCG)组合能够有效促使雌性花鳗鲡性腺发育成熟,GSI平均达22.9％,与对照组相比差异显著.单独注射雄烯二酮(ADSD)和注射ADSD、CPE、hCG混合制剂诱导效果不明显,GSI仅为1％ ～3％.荧光定量发现,经CPE与hCG处理的亲鱼LHβ和GtHα的表达量不断上调,其中发育早期GtHα的表达量极显著升高,而FSHβ的表达量呈现下调趋势.研究表明,CPE与hCG组合注射能够成功诱导花鳗鲡卵巢发育成熟,而ADSD对花鳗鲡卵巢发育无明显促进作用.LH在花鳗鲡卵巢发育整个过程中扮演重要角色;FSH在花鳗鲡性腺发育早期作用显著,可能是诱导鳗鲡性腺早期发育的关键因素.     

鲤、鲢、草鱼促性腺激素垂体含量及血清浓度日周期变化的关系

淡水鱼类有的在池塘中能自然产卵,如鲤(Gyprinus carpio)、鳊(Megalobrama amblyocephala)等,有的鱼必须给予外源激素才能产卵,如青鱼(Mylopharyngodon piceus)、草鱼(Ctenopharyngodon idellus)、鲢(Hypophthalmichthys molitrix)、鳙(Aristichthys nobilis)。为探索产生这种差别的原因,本文用放射免疫方法测定了鲤、鲢、草鱼产卵季节促性腺激素(GTH)在垂体中的含量和血清中浓度的日周期变化。鲤、草、鲢24小时血清GTH浓度呈脉冲式波动,白天出现3-4次高峰,夜间雌的有时还有一次高峰。鲤高峰值多在20-40毫微克／毫升血清,峰谷在5—15毫微克／毫升血清,GTH浓度始终保持高水平,而草、鲢高峰值仅2—8毫微克／毫升血清,峰谷0．1—1毫微克／毫升血清。草、鲢每只垂体GTH含量较高,约为鲤的2．5倍。以上数据说明草、鲢垂体释放GTH比鲤少。这种差别可能反映了在池塘中鲤能自产,而草、鲢不能自产的原因。草鱼垂体离体培养,白天和黑夜取材,对LRH-A的反应性不同,白天的反应较弱,对此进行了讨论。     

Direct actions of serotonin on gonadotropin-II and growth hormone release from goldfish pituitary cells: interactions with gonadotropin-releasing hormone and dopamine and further evaluation of serotonin receptor specificity

In this study, the direct actions of serotonin (5HT) on gonadotropin (GTH)-II and growth hormone (GH) release in the goldfish were tested at the pituitary cell level. 5HT (10 nM - 10 µM) stimulated GTH-II but inhibited GH release from perifused goldfish pituitary cells in a dose-dependent manner. The minimal effective dose of 5HT tested to suppress basal GH secretion (10 nM) was 10-fold lower than that to stimulate GTH-II release (100 nM). The GTH-II releasing effect of 5HT was abolished by repeated 5HT treatment (10 µM) whereas the corresponding inhibition on GH release was unaffected. These results suggest that 5HT receptors on goldfish gonadotrophs and somatotrophs exhibit intrinsic differences in terms of sensitivity to stimulation and resistance to desensitization. Salmon GTH-releasing hormone (sGnRH, 100 nM) stimulated GTH-II and GH release from goldfish pituitary cells. The GTH-II releasing action of sGnRH was unaffected by simultaneous treatment of 5HT (1 µM). However, the corresponding GH response to sGnRH (100 nM) was inhibited. In the goldfish, dopamine is known to stimulate GH release through activation of pituitary D1 receptors. In the present study, the GH-releasing action of dopamine (1 µM) and the D1 agonist SKF38393 (1 µM) was significantly reduced by 5HT (1 µM). To examine the receptor specificity of 5HT action, the effects of 5HT1 and 5HT2 analogs on GTH-II and GH release were tested in goldfish pituitary cells. The 5HT1 agonist 8OH DPAT (0.1 and 1µM) and 5HT2 agonist methyl 5HT (0.1 1µM) mimicked the GTH-II releasing effect of 5HT. The 5HT1 agonist 8OH DPAT (0.1 and 1µM) also stimulated GH release but the 5HT2 agonist methyl 5HT (0.1 and 1µM) was inhibitory to basal GH secretion. In addition, 5HT (1µM) -stimulated GTH-II release was abolished by the 5HT1 antagonist methiothepin (10µM) and 5HT2 antagonist mianserin (10µM). Similarly, the inhibitory action of 5HT (1µM) on basal GH release was blocked by the 5HT2 antagonist mianserin (10µM). The 5HT1 antagonist methiothepin (10µM) was not effective in this regard. These results, taken together, indicate that 5HT exerts its regulatory actions on GTH-II and GH release in the goldfish directly at the pituitary cell level, probably through interactions with other regulators including sGnRH and dopamine. The GTH-II releasing action of 5HT is mediated through 5HT2 and possibly 5HT1 receptors. The inhibition of 5HT on basal GH release is mediated through 5HT2 receptors only. Apparently, 5HT1 receptors are not involved in this inhibitory action. In this study, a paradoxical stimulatory component of 5HT on GH release by activating 5HT1 receptors is also implicated.     

Effects of photoperiod on gonadotropin-releasing hormone levels in the brain and pituitary of underyearling male barfin flounder

ABSTRACT:   A pleuronectiform fish, the barfin flounder Verasper moseri , expresses three gonadotropin-releasing hormone (GnRH) forms in the brain: salmon GnRH (sGnRH), chicken GnRH-II (cGnRH-II) and seabream GnRH (sbGnRH). To clarify the effects of photoperiod on GnRH systems, changes in brain and pituitary GnRH peptide levels were examined using time-resolved fluoroimmunoassays. In experiment 1, 5-month-old male barfin flounder (mean total length 9.0 cm, body weight 11.0 g) were divided into short (8:16 h light : dark [L:D] cycle; lights on 08.00–16.00 hours) and long photoperiod (16:8 h L:D cycle; lights on 04.00–20.00 hours) groups in mid September and maintained until November under natural water temperature (19.3–15.2°C). Brain sGnRH concentrations were significantly higher in the 16:8 h L:D group than in the 8:16 h L:D group, whereas no significant differences were observed in total length, body weight, plasma testosterone concentration, brain cGnRH-II concentration and pituitary sbGnRH content. In experiment 2, 7-month-old male barfin flounder (mean total length 16.5 cm, body weight 76.8 g) were divided into short and long photoperiod groups in mid December and maintained until February under natural water temperature (12.5–6.6°C). Total length, body weight and condition factor were significantly greater in the 16:8 h L:D group than in the 8:16 h L:D group, whereas no significant differences were observed in plasma testosterone concentration and GnRH levels in the brain and pituitary. These results indicate that levels of sGnRH in barfin flounder are influenced by photoperiodic treatment dependent on water temperature and/or body size.     

The regulatory effects of thyrotropin-releasing hormone on growth hormone secretion from the pituitary of common carp in vitro

The effects of thyrotropin-releasing hormone (TRH) on growth hormone (GH) and gonadotropin (GtH) release, and the influences of somatostatin (SRIF), the dopamine agonist apomorphine (APO) and extracellular calcium on basal and TRH-induced GH release were examined using an in vitro perifusion system for pituitary fragments of common carp (Cyprinus carpio). Five minute pulses of different dosages of TRH stimulated a rapid and dose-dependent increase in GH release from the perifused pituitary fragments with an ED₅₀ of 9.7 ± 2.3 nM. TRH was ineffective on GtH release. SRIF significantly inhibited basal and TRH-induced GH release from the perifused pituitary fragments, and the effects of SRIF were dose-dependent. APO induced a dose-dependent increase in basal and TRH-stimulated GH release from the perifused pituitary fragments. Increasing the concentrations of extracellular calcium from 0 mM to 1.25 mM resulted in an increase in basal and TRH-induced GH release. The high dose of calcium (6.25 mM) caused a slight decrease in basal and TRH-induced GH release compared with those at a concentration of 1.25 mM.

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Résumé Les effets de la thyrotropine (TRH) sur la sécrétion d'hormone de croissance (GH) et de gonadotropine (GTH), et de la somatostatine (SRIF), de l'apomorphine (APO), antagoniste dopaminergique, et du calcium extracellulaire sur les sécrétions basale et stimulée de GH ont été étudiées in vitro par périfusion, de fragments d'hypophyses de carpe (Cyprinus carpio). Des applications de 5 minutes de TRH à différentes concentrations induisent une stimulation rapide et dose dépendante de la sécrétion de GH (ED₅₀ = 9.7 ± 2.3 nM). Le TRH est sans effet sur la sécrétion de GTH. Le SRIF inhibe la sécrétion basale de GH ainsi que la résponse hypophysaire à l'action du TRH. Son action est dose dépendante. L'apomorphine induit une augmentation dose dépendante de la sécrétion basale de GH et potentialise l'action du TRH sur la stimulation de la sécrétion de GH. Des effets équivalents sont induits par des concentrations croissantes de calcium extra cellulaire de 0 à 1.2 mM, alors qu'à une concentration de 6.25 mM des effets opposés sont obtenus.

     

The effects of salinity on reproductive performance and plasma levels of sex steroids in Caspian kutum Rutilus frisii kutum

Sexually mature kutum, Rutilus frisii kutum, captured from its natural habits, the Caspian Sea and the Khoushkrood River, reared at 0.5 g L^?1 and 8–13 g L^?1 for approximately 1 year in experimental condition, for assessing the effect of salinity on reproduction. Plasma concentrations of sex steroid hormones (17β‐estradiol, testosterone and 17α‐hydroxyprogestrone) were measured in the three stages of gametogenesis. Female kutum held at <0.5 g L^?1 or 8–13 g L^?1 had no ovulated oocytes in their ovaries. In contrast, males held in captivity were spermiated, similar to their wild counterparts. The average sperm volume of males held at <0.5 g L^?1 (2.36 ± 0.46 mL) was lower than males held at 8–13 g L^?1 (3.65 ± 0.73 mL) at the end of the experimental period. The highest concentration of testosterone was observed in mid‐gametogenesis in wild fish that was significantly higher than the concentration seen in fish held in either <0.5 g L^?1 or 8–13 g L^?1. Female kutum showed suppressed steroid hormones in captivity, resulting in failure in the gonad development. However, male kutum adapt well to captivity and showed synchrony in steroid hormone variations with the wild fish, resulting in the testicular development. Results of this study also indicate that salinity plays a minor, but vital, role in reproduction of kutum, a factor that needs to be considered for keeping broodstock of brackish water fish species like kutum.     

注射T4对日本鳗鲡血清、卵、早期幼苗甲状腺激素含量变化及对幼苗存活率的影响

对产卵前的雌鳗注射了不同剂量的甲状腺素(T4),然后利用放射性免疫法(RIA)测定了雌鳗血清、卵、胚胎及幼苗各发育时间段三碘甲腺原氨酸(T3)和甲状腺素(T4)的含量变化。结果发现,注射外源T424h后,随着雌鳗血清中的T4含量达到峰值,T3含量也达到了峰值。说明日本鳗鲡体内存在5’-单脱碘酶。与此同时,随着血清中T3和T4含量达到峰值,卵中的T3和T4含量也达到了峰值,进而证明母体中的T3和T4能够转入鳗鲡幼苗。进一步调整T4的注射剂量和时间后发现,在21℃水温条件下,T4的剂量为10～15mg·(500g)-1时,注射催产针前12h注射的效果最好。用该方法注射T4后,幼苗各发育时间段的存活率与对照组比较存在显著差异,出膜第9天的存活率由对照组的1.3%提高到22.0%。但日本鳗鲡幼苗最终未能存活,说明甲状腺素并不是影响日本鳗鲡幼苗存活的唯一因素。     

Effect of temperature and salinity on sex inversion in Asian Seabass (Lates calcarifer): relationship with plasma sex steroids concentration and aromatase activity of gonad and brain

Asian Seabass which shows precocious sex inversion in cultured environment is a major impediment to maintain a proper broodstocks. Seabass grown commercially in freshwater were transported to the research facility and held in freshwater at 29°C for 4 weeks and acclimatized to the experimental conditions. Fish were daily fed with a commercial pellet (50% protein, 18 MJ kg^?1) to satiety. Blood, brain and gonad collected before and at the end of the experiment were analysed for sex steroids level and aromatase activity. There was an increase in Plasma E₂ levels with temperature in fish at 34°C whereas no significant difference was observed at 24 and 29°C although the highest plasma T level was detected in fish at 34°C which had significantly lower level than at the beginning, except those held at 24°C in freshwater. Plasma 11‐KT was significantly greater in fish at 24°C compared with 29 or 34°C which had clear opposite to the E₂. Aromatase activity in brain was higher at 29°C than at either 24 or 34°C, whereas gonadal aromatase recorded highest at 34°C. These findings concluded that high temperature in culture facility induces sex inversion of Asian Seabass.     

Influence of gonadotropic hormone-releasing hormone analog (GnRH-A) on plasma sex steroid profiles and milt production in male winter flounder,Pseudopleuronectes americanus (Walbaum)

The effects of gonadotropic hormone-releasing hormone analog (GnRH-A) treatment on the onset and duration of increases in plasma sex steroids and milt production (milt volume and number of spermatozoa) were investigated in prespawning male winter flounder. After treatment of maturing males during the winter with a single injection of either 20 or 200 g/kg [D-Ala⁶, Pro⁹-NHEt]LHRH (GnRH-A), plasma levels of testosterone and 11-ketotestosterone were increased within 12h and the steroid hormone levels remained elevated for long periods lasting several days. The androgenic steroid response of males was delayed after the administration of a lower dose of GnRH-A (2 g/kg). Although a single GnRH-A injection in December or January advanced the onset of spermiation in some males, only small amounts (<50 l) of milt could be collected. By March, all males were in spermiating condition following GnRH-A treatment; however, significant increases in sperm production, particularly increases in milt volume, occurred in fish twice treated with GnRH-A.Part of this work presented at the First Symposium of Fish Endocrinology, Edmonton, Alberta, Canada June 1988 and the Satellite Symposium on Applications of Comparative Endocrinology to Fish Culture, Almuñecar, Granada, Spain May 1989