| Project/Area Number |
05404056
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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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 | UNIVERSITY OF TOKYO |
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Principal Investigator |
TAKETANI Yuji Univ. of Tokyo, Obstetrics and Gynecology, Professor, 医学部・附属病院, 教授 (10114539)
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| Co-Investigator(Kenkyū-buntansha) |
FUJIWARA Toshihiro Univ. of Tokyo, Obstetrics and Gynecology, Research Associate, 医学部・附属病院, 助手 (80219063)
YANO Tetsu Univ. of Tokyo, Obstetrics and Gynecology, Assistant Professor, 医学部・附属病院, 講師 (50182390)
TSUTSUMI Osamu Univ. of Tokyo, Obstetrics and Gynecology, Associate Professor, 医学部・附属病院, 助教授 (60134574)
林 直樹 東京大学, 医学部(病), 助手 (60211489)
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| Project Period (FY) |
1993 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥26,700,000 (Direct Cost: ¥26,700,000)
Fiscal Year 1996: ¥4,200,000 (Direct Cost: ¥4,200,000)
Fiscal Year 1995: ¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1994: ¥7,700,000 (Direct Cost: ¥7,700,000)
Fiscal Year 1993: ¥11,500,000 (Direct Cost: ¥11,500,000)
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| Keywords | embryo / glucose / GLUT / activin / cryopersevation / epidermal growth factor / trophoblast / epidermal growth facter / 卵 / 初期胚 / グルコース / 子宮内膜 / 糖輸送担体 / 増殖因子 / cocyte / glucoce transporter / hexokinase / oocyte |
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| Research Abstract |
We developed some methods useful to investigate the mechanism of intrinsic activities in mammalian embryos : microculture technique, micro Western blot analysis, quantitative RT-PCR analysis and enzymatic cycling technique. By using these methods, we obtained a result that glucose incorporation is increasing during the development of preimplantation embryos, especially in the period when the embryos develop from two-cell stage to eight-cell stage. This phenomenon is based on the expression of glucose transporter (GLUT) expression, and it was also revealed that increasing of hexokinase activity during embryonic development has a great deal to do with this phenomenon.
We also observed the specificity of the embryos which developed in vitro, and the influence of some biochemical materials such as growth factors to these embryos. Delayd expresseion of hexokinase and GLUT in vitro causes inappropriate glucose incorporation during preimplantation embryo development which were largely correcte
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d by epidermal growth factor (EGF). The implantation rate in recipient mice of in vitro grown embryos is lower than in vivo grown ones, and treatment of embryos in vitro with EGF effectively prevents a decrease in implantation rate. In addition, we examined the effects of cryopreservation to the embryos. Freezing-thawing procedure of 2-cell embryos decreases not only the implantation rates to the recipients, but also decreases glucose incorporation of the blastocyst due to decreased expression of GLUT.This suggests that cryopreservation may have later consequences on the functional development of embryos.
In addition, an inmortalized human uterine endometrial cell line was established and characterized on the effect of estrogen and progesterone. Estrogen stimulated the proliferation and the expression of EGF in the cell line. The coculture system of the endometrial cell line and embryo can be used as in vitro model to investigate the interaction between endometrium and embryos during early embryonic development and implantation. Less
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