Search for genes involved in implantation and bioimaging of implantation aging GFP tagged mouse cells

胎盘在支持胎儿发育方面起着重要作用，如气体交换、营养供应和激素产生。胎盘缺陷是妊娠丢失和并发症的许多方面的基础；因此，了解和调节胎盘过程中的基因功能具有重要的临床意义。然而，缺乏一种简便有效的胎盘特异性基因操作方法阻碍了胎盘的研究。在寻找胎盘可视化方法的过程中，我们尝试将慢病毒（LV）载体应用于器官特异性基因操作。我们之前已经证明，用LV载体转导受精卵可以有效地产生转基因动物，然而转基因表达在胎儿和胎盘中都有发生。在这项资助下进行的研究中，我们用LV载体转导无带囊胚，期望胎盘特异性基因表达，因为大多数胎盘细胞是从囊胚最外层的滋养细胞分化而来的。由于我们使用了GFP作为转基因，所以我们用荧光显微镜观察转基因的表达。结果，在胚胎着床前的滋养细胞和整个妊娠期间的胎盘中都发现了GFP荧光。所有被分析的胎盘都携带转基因，而没有一个胎儿变成转基因。从而建立了胎盘可视化的基本方法。为了开发应用，我们进一步进行了实验。为了应用该方法，选择胚胎因胎盘缺陷而死亡的胚胎致死小鼠品系。在我们的实验中，我们可以证明，通过将转基因完全引入胎盘，几个基因敲除动物模型基本上得到了拯救。这项技术为胎盘器官发生的基因操作提供了一个强大的系统，对胎盘功能障碍基因治疗的未来临床应用具有重要意义。少

The placenta plays important roles to support fetal development, such as gas exchange, nutrient supply, and hormone production. Placental defects underlie many aspects of pregnancy losses and complications ; thus, understanding and regulating gene function during placentation is of high clinical relevance. However, the lack of a facile and efficient method for placenta-specific gene manipulation has hampered the study of placenta. During to find the way to visualize the placentation, we tried to apply the lentivirus (LV) vectors to organ specific gene manipulation. We have previously shown that the transduction of fertilized mouse eggs with LV vectors efficiently generates transgenic animals, however transgene expression occurred in both the fetus and the placenta. In our study performed under the support of this grant, we transduced zona-free blastocysts with LV vectors expecting placenta-specific gene expression, since most placental cells differentiate from trophoblast cells that fo … More rm the outermost layer of the blastocyst. Since we used GFP as a transgene, the transgene expression was observed by fluorescent microscopes. As a result, the GFP fluorescence was found in trophoblast cells from preimplantation stages and in placenta throughout gestation. All the analyzed placentas carried the transgene, while none of the fetuses became transgenic. Thus to establish the basic method to visualize the placenta was achieved. In order to develop the application use, we further performed experiments. For the application of this method, embryonic lethal mouse strains were chosen in which the embryos die by placental defects. In our experiment, we could demonstrate that several knockout animal models were substantially rescued by the introduction of transgene exclusively into the placenta. This technology provides a powerful system for gene manipulation exclusively in placental organogenesis with implications for future clinical usage for the gene therapy for placental dysfunction. Less