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A study on the regulatory mechanisms of early embryonic development and implantation.

早期胚胎发育与着床调控机制的研究。

基本信息

批准号：
05404056
负责人：
TAKETANI Yuji
金额：
$ 17.09万
依托单位：
UNIVERSITY OF TOKYO
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-05404056/
关键词：
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

项目摘要

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 … More 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

我们发展了一些有助于研究哺乳动物胚胎内在活性机制的方法：微培养技术、微Western blot分析、定量RT-PCR分析和酶循环技术。通过这些方法，我们得到了在着床前胚胎发育过程中葡萄糖掺入增加的结果，特别是在胚胎从二细胞期发育到八细胞期。这一现象是基于葡萄糖转运蛋白（GLUT）的表达，同时也揭示了胚胎发育过程中己糖激酶活性的增加与这一现象有很大关系。我们还观察了体外培养胚胎的特异性，以及生长因子等生化物质对体外培养胚胎的影响。体外己糖激酶和GLUT的延迟表达导致着床前胚胎发育过程中葡萄糖的不适当掺入，这在很大程度上是由表皮生长因子（EGF）纠正的。体外生长的胚胎在受体小鼠中的着床率低于体内生长的胚胎，体外用EGF处理胚胎可有效防止着床率的降低。此外，我们还研究了冷冻保存对胚胎的影响。2细胞胚胎的冻融过程不仅降低了受者的着床率，而且由于GLUT的表达减少，使囊胚的葡萄糖掺入减少。这表明低温保存可能会对胚胎的功能发育产生后续影响。此外，建立了人子宫内膜永生化细胞系，并对雌激素和黄体酮的作用进行了表征。雌激素可促进细胞增殖和EGF的表达。子宫内膜细胞系与胚胎共培养系统可作为研究胚胎早期发育和着床过程中子宫内膜与胚胎相互作用的体外模型。少