| Project/Area Number |
07807152
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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 UNIVERSITY,BRANCH HOSPITAL |
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Principal Investigator |
NISHII Osamu TOKYO UNIVERSITY,BRANCH HOSPITAL,DPT.OF OB/GYN,INSTRUCTOR, 医学部・附属病院(分), 助手 (40189254)
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| Co-Investigator(Kenkyū-buntansha) |
KOIZUMI Yoshio TOKYO UNIVERSITY,BRANCH HOSPITAL,DPT.OF OB/GYN,INSTRUCTOR, 医学部・附属病院(分), 助手 (90240720)
KATO Yoshio TOKYO UNIVERSITY,BRANCH HOSPITAL,DPT.OF OB/GYN,ASSISTANT PROF., 医学部・附属病院(分), 講師 (00185878)
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| Project Period (FY) |
1995 – 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: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1996: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1995: ¥600,000 (Direct Cost: ¥600,000)
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| Keywords | MULLERIAN INHIBITOR / GRANULOSA CELL / HUMAN / PT-PCR / QUANTATIVE RT-PCR / ミューレリアン インヒビター / 定量的 RT-PCR / 遺伝子発現 / ミューレリアン / インヒビター / RT-RCR |
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| Research Abstract |
The establishment of the system to detect mullerian inhibitor (MI) gene expression was tried using RT-PCR in primary cultured granulosa cells. Human granulosa cells were obtained by the aspiration of ovarian follicles from patients undergoing laparotomy. Imformed consent was obtained from the patients. The preparation of granulasa cells was done from follicular fluid aspirated and cultured in monolayr culture. Two to three days after, cells were collected and total RNA was extracted.
As preliminary experiments, different sets of primer were constructed and RT-PCR was tried using these primer sets. At last, it was demonstrated that RT-PCR by primer set at exon 3 and exon 4 gave stable and consistent amplified gene product.
At the next step, the regulatory mechanisms of MI gene expression in human granulosa cells were investigated by quantative RT-PCR.Beta-actin was used as internal control. Primary cultured granulosa cells were treated with estradiol (10^<-8>M), progesterone (10^<-8>M), 8-brome-cAMP (1microM), IL-1alpha (10^<-8>), IL-6 (10^<-8>M) for 12 hours. Quantative RT-PCR was done, using total RNA extracted from these cells. However, none of these substances did alter the MI gene expression.
These preliminary results suggest that other factors regulates MI gene expression in human granulosa cells. Futher study is required to elucidate the regulatory mechanism of MI gene expression.
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