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。     

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。     

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。     

鳗鲡繁殖生物学研究Ⅴ．性类固醇激素诱导雌鳗促性腺激素(GtH)分泌和卵巢发育的作用

多次注射雌二醇或睾酮对雌鳗促性腺激素(GtH)的合成与分泌活动以及卵巢发育都有明显的促进作用,其中睾酮的作用更较雌二醇显着;多次注射LHRH-A的促进作用则不明显.多次埋植雌二醇或睾酮都能促使脑垂体合成GtH,但所取血清样品中的GtH含量都未见明显增加;多次埋植雌二醇后血清中卵黄蛋白原的含量明显增加,但卵巢GSI增长幅度小,表明埋植雌二醇虽能促使肝细胞合成卵黄蛋白原并释放到血液中,但未能全部渗入卵母细胞内;多次埋植睾酮后血清中的卵黄蛋白原含量虽未见明显增加,但卵巢明显发育长大,这表明埋植睾酮能促使肝细胞合成的卵黄蛋白原迅速地通过血液运送到卵巢,并渗入卵母细胞内.       

多马胺能药物对鲇鱼促性腺激素（GtH）分泌活动的影响

以珠江流域鲇鱼（silurus asotus)为实验材料,研究了多巴胺（DA）能药物（DA及其D－2型受体拮抗物 ,DOM）对鲇鱼促性腺激素（GtH)释放的影响,结果表明,在性腺发育的各个时期,单独注射DOM（5ug/g)均不能显著提高鲇鱼血液基础GtH水平,当DOM与LHRH－A联合注射时能显著增强LHRH－A刺激GtH释放的作用;DA只能抑制GnRH诱导的GtH释放,对基础GtH释放无抑制作用,这种生殖内分泌调节方式与鲇形目的革胡子鲇（Clarias gariepinus)和大鳍Hu（Mystus macropterus)相似,而与鲤形目的鲁科（Cyrpindiae)鱼类不同。     

雄烯二酮和甲基睾酮诱导雄性日本鳗鲡性腺发育的作用

多次埋植雄激素雄烯二酮（ADSD）或甲基睾酮（MT）均可促进雄性日本鳗鲡（Anguilla japonica)性腺发育成熟,明显提高脑和垂体mGnRH,垂体GtH含量,埋植3次后,MT处理组的GSI及垂体GtH含量显著高于ADSD处理组,MT处理组血清GtH含量在第1次埋植后显著升高,而ADSD处理组在第4次埋植后才显贰高于对照组。这些结果表明：埋植ADSD和MT可反馈作用于雄性日本鳗鲡脑和垂体,促进GnRH和TtH的合成和分泌,进一步诱导精巢发育。而且MT的作用效果较ADSD快。     

鱼类促性腺激素分泌的调节机理和高效新型鱼类催产剂

鱼类和其他脊椎动物一样,由下丘脑—脑垂体—性腺轴调节整个生殖活动,其中由脑垂体合成与分泌的促性腺激素(GtH)起着十分重要的作用。在一定环境条件影响和下丘脑的调控作用支配下,使脑垂体GtH的合成与释放活动增强,它作用于性腺组织而诱导性类固醇激素的产生,进而促使配子发育成熟和排精与排卵。因此,长期以来都是使用外源的GtH,如鲤鱼(或其他鱼类)脑垂体匀浆液或粗提的人体绒毛膜促性腺激素(HCG)作为鱼类人工繁殖的催产剂。这些催产剂虽有较好效果,但来源有限,成本高,亲鱼易产生抗药性,催产后死亡     

池养鲤和草鱼血液学指标的研究

1.鲤鱼和草鱼的血液细胞包含红血球、淋巴球、单核球、嗜中性球、嗜酸性球及纺锤细胞。没有见到嗜碱性球。2.鲤鱼及草鱼的红血球数及血红蛋白量,在正常条件下,与水中溶氧量及成熟系数成十分显著的负相关。成熟雌鲤的红血球数(y)与溶氧量(x1)及成熟系数(x2)的迥归公式为:y=122+2.9x1-1.2x2。3.性腺发育程度及生殖活动强烈地影响鱼类血液有形成分。在生殖季节雄鱼的红血球数及血红蛋白量比雌鱼高,红血球沉降率则相反。白血球则在生殖季节及性腺退化吸收时比较活跃,而在非生殖季节两性之间无明显差异。4.发育成熟的雌鲤,经注射垂体后,无论产卵与不产卵其红血球数及血红蛋白量都下降,自然产卵的雌鲤亦然。5.雌鲤的红血球沉降率在临近产卵时急剧增加,而且比雄鲤明显地高。至于饥饿则对红血球沉降率无明显影响。6.在池养条件下鲤鱼的红血球数、血红蛋白量以及血式的个体差异较小,可望成为这类鱼的有应用价值的正常生理指标。       

利血平和丘脑下部促黄体素释放激素类似物(LHRH-A)对大鳞副泥鳅促性腺激素细胞的分泌活动和排卵的促进作用

利血平能使神经分泌细胞的儿茶酚胺贮存量减少甚至消耗殆尽,导致多巴胺作为GRIF的作用减弱,从而显著增强LHRH-A促进GtH分泌和诱导排卵的效应。对大鳞副泥鳅,利血平和LHRH-A同时注射或者利血平比LHRH-A提前3小时注射,都能激活GtH细胞,显著增强其分泌活动并诱导排卵。利血平的最低有效剂量为1—5微克/克体重。利血平(25微克/克体重) LHRH-A(0.05微克/克体重)的催产效果和多巴胺拮抗物pimozide(1微克/克体重) LHRH-A(0.05微克/克体重)的催产效果一样,排卵率都达到了100％。     

Testosterone and estradiol potentiate the serum gonadotropin response to gonadotropin-releasing hormone in goldfish

The effects of gonadal steroids on gonadosomatic index (GSI; gonad wt/total body wt x 100), pituitary gonadotropin (GTH) content, and serum GTH response to [D-Ala6,Pro9-Net]-luteinizing hormone-releasing hormone (LHRH-A) were investigated throughout the seasonal reproductive cycle of the goldfish. Gonad-intact female fish were implanted i.p. for 5 days with silastic pellets containing no steroid (blank), testosterone (T; 100 micrograms/g), or estradiol (E2; 100 micrograms/g). The serum GTH response at 6 h following i.p. injection of saline or 0.1 microgram/g LHRH-A was assessed. In blank-implanted, saline-injected animals, seasonal variations in GSI, pituitary GTH content, and serum GTH levels were evident; maximal and minimal levels were noted in the spring and summer months, respectively. In blank-implanted fish, LHRH-A effectively stimulated GTH release in females undergoing gonadal recrudescence (late autumn and winter) and in sexually mature (spring) females, but not in sexually regressed (summer and early autumn) females. Implantation of T or E2 raised serum steroid levels to those found during ovulation in goldfish. Steroid treatments did not affect unstimulated serum GTH levels at any time of the year. Testosterone effectively potentiated the serum GTH response to LHRH-A during the entire reproductive cycle, whereas the positive effects of E2 were evident in sexually regressed and post-spawning females only. Both T and E2 potentiated the GTH response to LHRH-A in male fish. To examine the involvement of T aromatization in mediating its actions on induced GTH secretion, male and female fish were implanted with T or the nonaromatizable androgens 5 alpha-dihydroxytestosterone (DHT; 100 micrograms/g) and 11-keto-testosterone (11-KT; 250 micrograms/animal). Testosterone potentiated the GTH response to LHRH-A in both males and females whereas DHT and 11-KT were without effect. Furthermore, the positive action of T on induced GTH secretion was blocked by 2-day pretreatment with the aromatase inhibitor 1,4,6-androstatrien-3,17-dione (100 or 300 micrograms/g). Multiple i.p. injections of hCG (0.2 microgram/g every 3 days for 39 days), probably through stimulation of endogenous T secretion, resulted in potentiation of the GTH response to LHRH-A in mature male goldfish. These results clearly demonstrate that T, through aromatization to E2, can increase pituitary responsiveness to exogenous LHRH-A in gonad-intact male and female goldfish.     

Precocious ovarian recrudescence and induced off–season spawning of Heteropneustes fossilis by D-Lys6 salmon gonadotropin releasing hormone analog (sGnRH-A)

The feasibility of photothermal treatment and use of salmon gonadotropin releasing hormone analogue (sGnRH-A) for induced spawning of Heteropneustes fossilis outside the spawning season was investigated. Fish were maintained at 30° C and 14 h light (L)/10 h dark (D) for 60 days between the post-spawning season and preparatory season. Fish attained gonadal maturity 4 months earlier compared to the natural spawning season. Mature fish were induced to spawn by D-Lys⁶ sGnRH-A at a dose of 25 μg kg⁻¹ body weight. The eggs obtained were fertilizable. Injection of D-Lys⁶ sGnRH-A at different time points during the photodiurnal cycle did not influence the latency period and spawning of H. fossilis .     

Reproduction and spawning in walleye (Stizostedion vitreum)

The walleye ( Stizostedion vitreum ) is a North American mesothermal freshwater teleost that spawns once each year in early spring. Walleye spawn randomly over suitable substrates and do not provide any parental protection for eggs or juveniles. The majority of gonadal recrudescence in adult male walleye occurs in the autumn, and walleye testes contain large numbers of viable spermatozoa from late autumn through the spawning season. Adult female walleye exhibit group synchronous ovarian development, and similar to males, the majority of gonadal development occurs in the autumn. Evidence suggests that 17α,20β-dihydroxyprogesterone is the maturational steroid in this species. Simple environmental manipulations coupled with injections of human chorionic gonadotropin can be used to advance spawning in walleye by up to 12 weeks. To spawn and propagate walleye, hatcheries in North America use a wide range of methods that have been developed to meet the needs and conditions present at specific facilities.     

The induced spawning of roach, Rutilus rutilus (L.), with the antioestrogens clomiphene and tamoxifen

The induced spawning of gravid roach was investigated using the antioestrogens clomiphene and tamoxifen. Three dose levels ofeach were used: 0·1, l or 10 mgkg^-1, given twice with a 4-day interval to groups of eight fish with saline controls. Running males were randomly distributed. Tamoxifen, when injected at a rate of l mg kg^-1 was found to be most successful. This treatment induced ovulation in five of the six females, and profuse spawning on 3 consecutive days, 4 days after the first injection. Clomiphene induced ovulation and spawning in one of six females at 2 × 10 mg kg^-1, and two of eight females at 2 × l mg kg^-1, respectively 6 and 7 days after the first injection. The eggs produced showed normal devel-opment. No control fish ovulated or spawned. Both drugs probably act by indirect mechanisms, blocking sex steroid feedback inhibition of gonadotropin (GtH) secretion at the pituitary, thereby inducing a plasma GtH surge. The results of this experiment suggest that tamoxifen may be an effective substitute for pituitary preparations in the induced spawning of fish.     

Effects of short periods of warm water fluctuations on reproductive endocrine axis of the pejerrey (Odontesthes bonariensis) spawning

The aim of this study was to assess fluctuations in daily water temperature in Chascomús Lagoon during one year, and to evaluate whether the highest temperature recorded during pejerrey spawning season can produce an endocrine disruption on brain-pituitary-gonads axis. Fish were subjected to daily temperature fluctuations: 17 °C to 19 °C (reproductive control), 19 °C to 25 °C, and 19 °C to 27 °C. After 8 days, ten fish per treatment were sacrificed and gene expression of gonadotropin-releasing hormone (GnRH-I, GnRH-II, GnRH-III), gonadotropin subunits-β (FSH-β, LH-β), glycoprotein hormone-α (GPH-α), gonadotropin receptors (FSH-R, LH-R), and gonadal aromatase (cyp19a1a) was analyzed. Also, plasma levels of sexual steroids and gonadal reproductive status were studied. Fish exposed to high temperature fluctuations quit spawning, presenting clear signs of gonadal regression. Fish recovered its spawning activity 11 weeks after heat treatment. At endocrine level, GnRH-I and FSH-β in both sexes, LH-β and GPH-α in males and FSH-R, LH-R and cyp19a1a in females decreased significantly in treated fish. Also, a strong reduction in plasma sex steroid levels was found for both sexes. This study demonstrated that pulses of warm water in natural environment during pejerrey spawning season can disrupt all levels of the reproductive axis, impairing reproduction.     

Estradiol-17β effects on lipid and carbohydrate metabolism and on the induction of a yolk precursor in goldfish,

The effects of estradiol-17β treatment on plasma lipid levels, liver lipid and glycogen reserves were examined during different phases of the reproductive cycle in goldfish, . Estrogen therapy resulted in increased plasma and hepatic lipid levels except during the spawning season. Hepatic glycogen deposits were depleted by estradiol injections during all seasons. Treatment of fish with the estrogen antagonist, CI-628, during the spawning season caused a reduction in plasma and liver lipid levels. Electrophoretic studies conducted during the post-spawning season showed that estrogen induces the appearance of a specific lipoprotein, probably a yolk precursor, in the serum and liver of goldfish.     

Time- and dose-related effects of gonadotropin-releasing hormone on growth hormone and gonadotropin subunit gene expression in the goldfish pituitary

The goldfish brain contains two molecular forms of gonadotropin-releasing hormone (GnRH): salmon GnRH (sGnRH) and chicken GnRH-II (cGnRH-II). In a preliminary report, we demonstrated the stimulation of gonadotropin hormone (GtH) subunit and growth hormone (GH) mRNA levels by a single dose of GnRH at a single time point in the goldfish pituitary. Here we extend the work and demonstrate time- and dose-related effects of sGnRH and cGnRH-II on GtH subunit and GH gene expression in vivo and in vitro. The present study demonstrates important differences between the time- and dose-related effects of sGnRH and cGnRH-II on GtH subunit and GH mRNA levels. Using primary cultures of dispersed pituitary cells, the minimal effective dose of cGnRH-II required to stimulate GtH subunit mRNA levels was found to be 10-fold lower than that of sGnRH. In addition, the magnitudes of the increases in GtH subunit and GH mRNA levels stimulated by cGnRH-II were found to be higher than the sGnRH-induced responses. However, no significant difference was observed between sGnRH and cGnRH-II-induced responses in vivo. Time-related studies also revealed significant differences between sGnRH- and cGnRH-II-induced production of GtH subunit and GH mRNA in the goldfish pituitary. In general, the present study provides novel information on time- and dose-related effects of sGnRH and cGnRH-II on GtH subunit and GH mRNA levels and provides a framework for further investigation of GnRH mechanisms of action in the goldfish pituitary.