MATURATION AND SPAWNING OF WILD TIGER PUFFER TAKIFUGU RUBRIPES AND MECHANISM OF MATURATION ELUCIDATED BY CULTURED ONE

1995年：为了给水产养殖提供受精卵，我们建立了用激素诱导河豚卵母细胞成熟排卵的有效方法。LHRH-a胆固醇颗粒(LHRH-a 400 mg/kg)成功地诱导了卵黄积聚卵巢的3岁养殖河豚的卵母细胞过度成熟。随后，单次注射人绒毛膜促性腺激素(500IU/kg)和鲑鱼脑垂体(SP)匀浆(7 mg/kg)，以促进鱼类排卵。在实验I(卵母细胞直径900 mm的鱼)中，从7条鱼中有5条获得受精卵。实验二(卵母细胞直径在800~900 mm之间的鱼)，10尾鱼中有8尾获得受精卵。相反，在实验三中，按常规方法(每隔5天重复注射HCG+SP，不使用LHRH-a处理)，5条鱼中有2条获得了受精卵。平均费尔蒂…三次实验的复发率分别为55.1%、53.6%和11.0%。在实验中I和II，排卵主要发生在注射HCG+SP后96~120h。因此，开发了一种从养殖的河豚中获得高质量卵的技术。1996：本研究旨在为河豚真正的成熟诱导激素(MIH)提供直接证据。为此，我们在卵母细胞发育的四个不同阶段使用了无卵黄滤泡细胞。去卵黄卵泡细胞与~(14)C标记的17a-羟孕酮孵育2小时，然后用层析法分离放射性类固醇。对产生的放射性代谢产物进行鉴定，并测试其在体外诱导卵母细胞生发泡破裂(GVBD)的能力。此外，通过监测最终卵母细胞成熟期间的血清类固醇水平，研究了MIH候选者的体内产生情况。本研究结果为20b-S为河豚体内天然存在的MIH提供了证据。较少

1995 : We developed the effective method of induction of ovarian maturation and ovulation of the cultured tiger puffer Takifugu rubripes by hormonal treatments in order to supply the fertilized eggs for aquaculture. The LHRH-a cholesterol pellet (LHRH-a 400 mg/kg) implantation successully induced overian maturation for three-year-old cultured tiger puffer which had yolk accumulated ovaries. Thereafter single injection of HCG (500IU/kg) combined with chum salmon pituitary (SP) homogenate (7 mg/kg) was applied to the fish to sinchronize ovulation among fish. In Experiment I (fish with oocytes *900 mm in diameter), fertilized eggs were obtained from 5 of 7 fish used. In Experiment II (fish with oocytes between 800 and 900 mm in diameter), fertilized eggs were obtained from 8 of 10 fish tereated. On the contrary, in Experiment III as a conventional method (repeated injection of HCG+SP at 5 days interval without LHRH-a treatment), fertilized eggs were obtained from two of 5 fish. Mean ferti … More lization rates in the three experiments were 55.1,53.6, and 11.0%, respectively. In Exp. I and II,ovulation was mainly occurred between 96 and 120 hours after HCG+SP injection. Thus, a thchnique for obtaining eggs of high quality from the cultured tiger puffer was developed.1996 : The study was designed to obtain derect evidence for the authentic maturation-inducing hormone (MIH) of tiger puffer, Takifugu ruberipes. To this end, we used yolk free follicle cells at four different stages of oocyte development. The yolk free follicle cells were incubated with the ^<14>C-labeled 17a-hydroxyprogesterone for 2hr, followed by chromatographic separation of the radioactive steroids. The radioactive metabolites produced were identified and tested for their ability to induce germinal vesicle breakdown (GVBD) in oocytes in vitro. Furthermore, in vivo production of candidates for MIH was investigated by monitoring the serum steroid levels during final oocyte maturation. The results obtained in this study provide evidence that 20b-S is the naturally occurring MIH in tiger puffer. Less