| Project/Area Number |
08671874
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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 |
Obstetrics and gynecology
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| Research Institution | Tokyo Medical & Dental University, Medical Research Insitute |
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Principal Investigator |
ASAKAI Rei Tokyo Med.& Dent.Univ., Medical Res.Inst., Res.Assistant, 難治疾患研究所機能調節疾患(内分泌), 助手 (20167224)
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| Project Period (FY) |
1996 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1997: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1996: ¥900,000 (Direct Cost: ¥900,000)
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| Keywords | granulosa cells / FGF receptor / Ad4BP / FSH / LH / AD4BP / FGF-2レセプター / プロテインキナーゼC |
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| Research Abstract |
(i) Basic firbroblast growth factor (FGF2) receptor (FGFR) on cultured porceine granulosa cells was found to be regulated by EGF ; the receptor down-regulation by EGF is comprised of the acute (within 30 minutes) phase and the following chronic phase. Both phases are due to PKC activation since a PKC inhibitor inhibits these two phases. Diacylglycerol (DAG) formation was observed at 2 min, 15-20 min, and 1 hour after EGF stimulation. The DAG at 2 min was acompanied by inositol triphosphate production. In the presence of 1% ethanol phosphatidyl ethanol formation was observed at 30 min after EGF stimulation, indicating the involvement of phospholipase C or D.Collectively these data suggest the involvement of several species of phospholipases in the FGFR downregulation.
(ii) A cell line for granulosa cells was extablished, since primary cultured cells have many limitations in molecular analyzes of FGFR.The transfection of mouse granulosa cells with SV40T antigen and AD4BP cDNA gave a birth to an unique cell line (4B2) that apparently mimics primary cultured granulosa cells ; 1) cAMP stimulates progesterone secretion, and induces morphological changes as well as growth arrest, and 2) both FSH and LH are able to stimulate proliferation of the cell line. In experiments of tumor formation in SCID mice, 2 months after cell implantation 2 to 5 mm in diameter of tumors are formed, whose morphologies are found to be similar to human granulosa cell tumor although the tumors secrete progesterone but not estradiol.
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