| Project/Area Number |
62570758
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Obstetrics and gynecology
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| Research Institution | Kobe University |
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Principal Investigator |
ASHITAKA Yoshihiko Kobe University School of Medicine Dept. of Obstetrics and Gynecology, Associate Professor, 医学部, 助教授 (10030959)
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| Co-Investigator(Kenkyū-buntansha) |
MARUO Takeshi Kobe University School of Medicine Dept. of Obstetrics and Gynecology, Assistant, 医学部付属病院, 講師 (60135811)
MOCHIZUKI Matsuto Kobe University School of Medicine Dept. of Obstetrics and Gynecology, Professor, 医学部, 教授 (80030922)
上田 康夫 神戸大学, 医学部, 助手 (20168636)
西村 隆一郎 神戸大学, 医学部, 助手 (00127383)
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| Project Period (FY) |
1987 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1989: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1988: ¥200,000 (Direct Cost: ¥200,000)
Fiscal Year 1987: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | Thyroid Hormone / Nuclear Receptor / Human Placental Trophoblast / cytotrophoblast / sycytiotrophoblast / Functional Differentiation / Cell Culture / Granulosa Cell / 甲状線ホルモン / 核レセプター / ヒト胎盤絨毛細胞 / ブタ顆粒膜細胞 / 抗核レセプター抗体 / ヒト胎盤繊毛細胞 / 核蛋白 / 機能文化 / steroidogenesis / aromatization |
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| Research Abstract |
1.Studies on thyroid hormone receptor (NT_3R) in human placental trophoblast. (1) Studies on the nuclear 3,5,3 -triiodo-L-thyronine binding sites in human placental trophoblast. In a previous study, we demonstrated that human term placental trophoblast contained NT_3R. In the first and second trimesters trophoblast, association constant(Ka) was 6.1 <plus-minus> 1.2 X 10^<-9>M, the capacity(Ca) was 105.6 <plus-minus> 59.9 fmol T_3/mg DNA. In term, however, Ka was 6.8 <plus-minus> 1.0 X 10^<-9>M and Ca was 63.8 fmol T_3/mg DNA. There was no difference in biochemical characterisity of both early and term placenta. But Ca would trend to be decreasing toward term. On the other hand, Ka in cytotrophoblast was 5.2 x 10^<-9>M, and Ca was 445 fmol T_3/mg DNA. These findings indicated that cytotrophoblast is most likely the target cell of thyroid hormone. Furthermore, optimal T_3,concentration (10^<-8>M) promoted hCG secretion in trophoblast culture system. (2)Purification of NT_3R and preparation of monoclonal anti-NT_3R antibody. Nuclear protein which was extracted from trophoblast by 0.4M KC1 was partially purified by DEAE Sephadex A 50 chromatography, used to immunized mice, and we have prepared polyclonal and monoclonal anti-NT_3R antibodies obtained from hybridoma cells. At present there are some problems in selecting those antibodies. According to SDS PAGE analysis of NT_3R cobalent bound with ^<125>I-T_3 PAL by applying ultraviolet lump, it would be cleared that the molecular weight of NT_3R was 46Kd.
2.Biocellular effect of thyroid hormone on functional differentiation of porcine granulosa cells in culture. It indicated that concomitant treatment with thyroid hormone (optimal concentration of 10^<-8>M T_3) promoted FSH-dependent morphological luteneization, i.e. alteration of immature granulosa cells to epithelioid form. Forethermore, it promoted aromatization in steroido- genesis and increased FSH-stimulated induction of hCG/LH receptor.
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