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In-vivo gene transfer into endometrial epithelial cells and endometrial stromal cells during pen-implantation period

笔植入期间体内基因转移至子宫内膜上皮细胞和子宫内膜基质细胞

基本信息

批准号：
12671591
负责人：
MINOURA Hiroyuki
金额：
$ 2.05万
依托单位：
Mie University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671591/
关键词：
embryonal implantation endome trium specific gene expression gene transfer endometrial epithelial cells enometrial stromal cells

项目摘要

The greatly increased knowledge on preimplantation embryos and the molecular biology of the uterus have led to the development of new concepts on the implantation process. However, the exact molecular events in the endometrium induced by the embryonal implantation are still far from clear. It is important to identify the factors participating in the feto-maternal relationships around the time of embryo implantation. We identified up-or down regulated genes, such as LRG-21, neuropeptide Y, Ghrelin during embryonal implantation in mouse uterus using differential display method, subtraction RT-PCR method and gene tip method. To clarify the functions of these genes, we attempted to establish the method of in-vivo gene transfer into endometnial epithelial cells and endometrial stromal cells during pen-implantation period. We used reporter gene constructs to transfect mouse endometnium in vivo. We injected DNA-liposome complexes into the uterine lumen, endometnium and tail vessel of mice on day 0.5 〜5 of pregnancy and detected reporter gene activity 2 days later. Unfortunately, incorporation rates of plasmid into endometnial epithelial cells and endometnial stromal cells were less than 10%.

对植入前胚胎和子宫分子生物学的了解大大增加，导致了植入过程新概念的发展。然而，胚胎植入引起的子宫内膜确切的分子事件仍远不清楚。确定胚胎植入期间影响母婴关系的因素非常重要。我们利用差异显示法、差减RT-PCR法和基因提示法鉴定了小鼠子宫胚胎着床过程中上调或下调的基因，如LRG-21、神经肽Y、Ghrelin。为了阐明这些基因的功能，我们尝试建立在笔植入期间将基因体内转移到子宫内膜上皮细胞和子宫内膜基质细胞的方法。我们使用报告基因构建体在体内转染小鼠子宫内膜。我们在妊娠第0.5〜5天将DNA-脂质体复合物注射到小鼠的子宫腔、子宫内膜和尾血管中，并在2天后检测报告基因活性。不幸的是，质粒进入子宫内膜上皮细胞和子宫内膜基质细胞的掺入率低于10%。