STUDY ON TEH MECHANISM OF GONADOTROPIN SYNTHESIS IN JAPANESE EEL

一个敏感的定量系统，使用逆转录-聚合酶链反应，以测量垂体GTH I β和II β的mRNA水平在雌性日本鳗鱼人工成熟诱导鲑鱼垂体匀浆（SPH）多次注射。注射SPH后，GTH I β mRNA水平降低，而GTH II β mRNA水平升高。使用GTH I β和II β的寡肽抗体的免疫组织化学分析也支持这一结果。此外，还建立了鳗鱼垂体器官培养体系，以分析GTH I β和II β表达的调控机制。在存在或不存在雌二醇-17 β（E2）的情况下，将来自未成熟雄性的分离垂体孵育1周。在E2孵育的垂体中观察到大量的GTH Ⅱ细胞，而对照组中则很少。为了测量孵育培养基或血液中GTH I和II的浓度，使用大肠杆菌产生重组GTH I β和II β并纯化作为标准品。然而，建立一个灵敏、特异的完整GTH I和II的定量体系仍需进一步研究。

A sensitive quantitation system using reverse transcription-polymerase chain reaction was developed to measure pituitary GTH Ibeta and IIbeta mRNA levels in female Japanese eel where artificial maturation was induced by multiple injections of salmon pituitary homogenate ( SPH). The GTH Ibeta mRNA levels decreased after SPH injections, while the GTH IIbeta mRNA levels increased. Immunohistochemical analysis using oligopeptide antibodies for GTH Ibeta and IIbeta also supported this result. In addition, an organ culture system for eel pituitary was developed to analyze the mechanisms for the regulation of GTH Ibeta and IIbeta expressions. Isolated pituitaries from immature males were incubated for 1 week in the presence or absence of estradiol-17beta (E2 ). A large number of GTH II cells were observed in the pituitary incubated with E2.In contrast, few GTH II cells were found in controls. To measure concentrations of GTH I and II in incubation media or blood, recombinant GTH Ibeta and IIbeta using E.coli were produced and purified as standards. However, establishment of a sensitive and specific quantitation system for intact GTH I and IIstill requires further research.