Contribution of Collagen to the development of mouse early embryo

胶原蛋白对小鼠早期胚胎发育的贡献

基本信息

批准号：
10660276
负责人：
TOMOGANE Hiroshi
金额：
$ 2.05万
依托单位：
Nippon Veterinary and Animal Science University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

项目摘要

Study(1) : A fertilized ovum repeats cell divesion cycles to increase the number of cell. The increased cell forms morula and blastocyst. In all tissues there is always collagen as extracellular matrix. To determine the contribution of collagen to the development of preimplantation embryos, cis-4-hydroxy-2-proline (CHP), collagen sysnthesis inhibitor, was added into the medium for the in vitro development of mouse 2-cell embryo and the in vitro development of 2-cell stage to blastocyst was estimated by the developmental rate to 4-cell, 8-cell, morula and blastocyst and by the number of cell at the developed blastocyst. The CHP addition blocked to develop to 4-cell satge from 2-cell stage. This blocking effect was inhibited by the replacement of proline at the same of CHP addition but not the replacement of lysine or glycine. Study(2) : The blocking effect of CHP on in vitro embryonic development was examined at various stages of embryos. The blocking occurred remarkably at the stages of from 2-cell to 4-cell and from morula to blastocyst. Study(3) : Using reverse transeription polymerase chain reaction method, mRNA of type I and III collagen weakly appeared at the stage of 4-cell. The typelV collagen mRNA first appeared from 8-cell stage and more at the blastocy. Study(4) : The collagen at the various stages of preimplantation embryos was determined using immuno-fluoresence- histochemistry. The collagen immuno-reaction was weakly detected at 2-cell, 4-cell and 8-cell stages and strongly at both morula and blastocyst cmbryos. These results indicate that collagen has important role for the early embryonic development and is essential subustance to develop to 4-cell stage and to blastocyst from morula.

研究（1）：施肥的卵子重复细胞潜水循环以增加细胞数量。增加的细胞形成了毛拉和胚泡。在所有组织中，总有胶原蛋白作为细胞外基质。 To determine the contribution of collagen to the development of preimplantation embryos, cis-4-hydroxy-2-proline (CHP), collagen sysnthesis inhibitor, was added into the medium for the in vitro development of mouse 2-cell embryo and the in vitro development of 2-cell stage to blastocyst was estimated by the developmental rate to 4-cell, 8-cell, morula and blastocyst and by the number of cell在开发的胚泡。 CHP的添加剂从2细胞阶段开发至4细胞SATGE。在添加CHP的同一脯氨酸上替代脯氨酸，而不是替代赖氨酸或甘氨酸，可以抑制这种阻断效果。研究（2）：在胚胎的各个阶段检查了CHP对体外胚胎发育的阻塞效应。封锁发生在从2细胞到4细胞的阶段以及从羊肚菌到胚泡的阶段。研究（3）：使用反向透射聚合酶链反应方法，I型和III胶原蛋白的mRNA弱出现在4细胞的阶段。 Typelv胶原mRNA最初是从8细胞阶段出现的，在胚泡中更多。研究（4）：使用免疫荧光组织化学确定植入前胚胎的各个阶段的胶原蛋白。在2细胞，4细胞和8细胞阶段，胶原蛋白免疫反应被弱检测到，在莫拉和胚泡cmbryos上都强烈地检测到。这些结果表明，胶原蛋白在早期的胚胎发育中具有重要作用，并且是发展为4细胞阶段并从毛拉的胚泡的必不可少的。