1989 Fiscal Year Final Research Report Summary
Studies on the Thyroid Hormone Actions in the Field of Reproduction
| 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)
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| Project Period (FY) |
1987 – 1989
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| Keywords | Thyroid Hormone / Nuclear Receptor / Human Placental Trophoblast / cytotrophoblast / sycytiotrophoblast / Functional Differentiation / Cell Culture / Granulosa Cell |
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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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[Publications] Maruo,T.,Hayashi,M.,Matsuo,H.,Yamamoto,T.,Okada,H.,Mochizuki,M.: "The role of thyroid hormones as a biological amplifier of folliclestimulating hormone in the function of cultured porcine granulosa cells." Endocrinology. 121. 1233-1241 (1987)
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「研究成果報告書概要(和文)」より
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[Publications] Ashitaka,Y.,Maruo,M.,Takeuchi,Y.,Nakayama,H.,Mochizuki,M.: "3,5,3'-triiodo-L-thyronine binding sites in nuclei of human trophoblastic cells." Endocrinologia Japonica. 35. 197-206 (1988)
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「研究成果報告書概要(和文)」より
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[Publications] Nishii,H.,Ashitaka,Y.,Maruo,M.,Takeuchi,Y.,Nakayama,H.,Mochizuki,M.: "Studies on the binding sites of 3,5,3'-triiodo-L-thyronine in cytotrophoblast." Endocrinologia Japonica. 36. (1989)
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[Publications] Ashitaka,Y.,Maruo,M.,Takeuchi,Y.,Nakayama,H.and Mochizuki,M.: "Human Placenta is a target organ of thyroid hormones.in“HUMAN REPRODUCTION-Current Status and Future Prospect"" Excerpta Medica,Amsterdam-New York-Oxford, 822-824 (1988)
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[Publications] Katayama,K.,Maruo,M.,Ashitaka,Y.,Mochizuki,M.et al.: "The significance of thyroid hormones in the maintenance of pregnancy.in“HUMAN REPRODUCTIONーCurrent Status and Future Prospect"editioned by Iizuka,R.and Semm.K." Excerta Media,Amsterdam-New York-Oxford, 597-598 (1988)
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