1998 Fiscal Year Final Research Report Summary
STUDY ON TEH MECHANISM OF GONADOTROPIN SYNTHESIS IN JAPANESE EEL
| Project/Area Number |
09660189
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General fisheries
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
ADACHI Shinji Fac.of Fish., Hokkaido Univ.Asso.Pro., 水産学部, 助教授 (40231930)
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| Project Period (FY) |
1997 – 1998
|
| Keywords | EEL / GONADOTROPIN / PITUITARY / GTH / RT-PCR / ORGAN CULTURE / IMMUNOHISTOCHEMISTRY / ARTIFICIAL MATURATION |
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| Research Abstract |
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 II
still requires further research.
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