Project/Area Number |
14540620
|
Research Category |
Grant-in-Aid for Scientific Research (C)
|
Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
生物形態・構造
|
Research Institution | Okazaki National Research Institutes |
Principal Investigator |
YOSHIKUNI Michiyasu Okazaki National Research Institutes, National Institute for Basic Biology, Lab.Reproductive Biology, Associate professor, 基礎生物学研究所, 助教授 (50210662)
|
Project Period (FY) |
2002 – 2003
|
Project Status |
Completed (Fiscal Year 2003)
|
Budget Amount *help |
¥4,100,000 (Direct Cost: ¥4,100,000)
Fiscal Year 2003: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2002: ¥2,400,000 (Direct Cost: ¥2,400,000)
|
Keywords | oocyte maturation / Olyzias latipes / steroid hormone / membrane receptor / gene cloning / non-genomic / ステロイドホルモン / 細胞受容体 / 細胞膜受容体 / GTP結合蛋白質 / 卵母細胞 / 情報伝達 |
Research Abstract |
In this study, we succeeded to isolate two cDNA clones of maturation-inducing steroid hormone (17alpha,20beta-dihyciroxy-4-pregnen-3-one) receptors, from medaka (Olyzias la tipes) ovary. These two cDNAs are classified as alpha (OlGPCPR-alpha) and gamma (OlGPCPR-gamma) subtypes. The cDNAs of OlGPCPR-alpha and OlGPCPR-gamma are 2983 by and 1566 by in lengths, respectively, and both cDNAs have an open reading frame encoding a protein of 352 amino acids. Sequence analysis revealed that both belonged to the G-protein-coupled receptor family with the typical 7 transmembrane domains. The amino acid sequence of OlGPCPR-alpha has 86.9% identity with the seatrout sequence and 48.4% with OlGPVPR-gamma. Northern blot analysis shows that OlGPCPR-alpha is expressed in brain, heart, kidney, ovary and testis, in contrast, OlGPCPR-gamma is expressed in ovary and testis. Polyclonal antibodies were generated in rabbits for Western blotting analysis of the receptor proteins. We also isolated a cDNA clone of the medaka homologue of membrane progestin receptor with a single transmembrane domain. The cDNA has an ORF encoding a protein of 198 AA and has 52 % identity with the sequence of human DG6.
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