Project/Area Number |
07557256
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Research Category |
Grant-in-Aid for Scientific Research (A)
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Allocation Type | Single-year Grants |
Section | 試験 |
Research Field |
Obstetrics and gynecology
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Research Institution | UNIVERSITY of TOKYO |
Principal Investigator |
TAKETANI Yuji Univ. of Tokyo, Obotetrcs and Gynecology, Professor the embryonic fanction., 医学部附属病院, 教授 (10114539)
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Co-Investigator(Kenkyū-buntansha) |
INOUE Satoshi Univ. of Tokyo, geriatrics, Reseach Associate, 医学部附属病院, 助手 (40251251)
MURAMATSU Masami Saitama Medical Univ., Biochemics II,Professor., 医学部附属病院, 教授 (10035454)
TSUTSUMI Osamu Univ. of Tokyo, Obotetrcs and Gynecology, Associate Professor, 医学部附属病院, 助教授 (60134574)
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Project Period (FY) |
1995 – 1996
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Project Status |
Completed (Fiscal Year 1996)
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Budget Amount *help |
¥5,400,000 (Direct Cost: ¥5,400,000)
Fiscal Year 1996: ¥5,400,000 (Direct Cost: ¥5,400,000)
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Keywords | embryo / glucose / GLUT / activin / cryopersevation / epidermal growth factor / trophoblast / embryo / epiclermal growth factor / 卵 / 初期胚 / 糖輸送担体 / 細胞増殖因子 / グルコース取り込み / 受精 / 着床 |
Research Abstract |
We developed some methods that are very 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 obseived the specificity of the embryos which developed in vitro, and the influence of some biochemical materials such as growth factors to these embryos. Delayd expression of hexokinase and GLUT in vitro causes inappropriate glucose incorporation during preimplantation embryo development which were largely corrected 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. From these facts we conclude that glucose incorporation in embryos based on the expression of hexokinase and GLUT can be a measure of embryo viability.
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