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

寻找参与植入老化 GFP 标记小鼠细胞植入和生物成像的基因

• 批准号: 17300135
• 负责人: OKABE Masaru
• 金额: $ 10.21万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17300135/
• 关键词: bioima ging; placenta specific gene manipulation; lentivirus; implantation; giant cells; fusion; syncytin; レンチウィルス; GFP; 精子; 卵子; 受精; 胎盤; 遺伝子導入; 蛍光たんぱく質; 子宮; 輸卵管

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

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。胎盘缺陷是妊娠损失和并发症的许多方面的基础;因此，了解和调节胎盘形成过程中的基因功能具有高度的临床意义。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。在寻找胎盘形成可视化方法的过程中，我们尝试将慢病毒载体应用于器官特异性基因操作。我们以前已经证明，用LV载体转导受精小鼠卵可以有效地产生转基因动物，然而转基因表达发生在胎儿和胎盘中。在我们的研究中，在该基金的支持下，我们用LV载体转导无透明带囊胚，期望胎盘特异性基因表达，因为大多数胎盘细胞从滋养层细胞分化而来， ...更多信息 rm囊胚的最外层。由于我们使用GFP作为转基因，因此通过荧光显微镜观察转基因表达。结果，GFP荧光在着床前阶段的滋养层细胞和整个妊娠期的胎盘中被发现。所有分析的胎盘都携带转基因，而没有一个胎儿成为转基因。从而建立了胎盘可视化的基本方法。为了开发应用程序使用，我们进一步进行了实验。对于该方法的应用，选择胚胎致死小鼠品系，其中胚胎因胎盘缺陷而死亡。在我们的实验中，我们可以证明，通过将转基因专门引入胎盘，几种基因敲除动物模型基本上得到了拯救。这项技术提供了一个强大的系统，专门在胎盘器官发生的基因操作的影响，为未来的临床使用的基因治疗胎盘功能障碍。少

项目成果

Observation of sperm-egg interaction through gene-manipulated animals

通过基因操作动物观察精卵相互作用

• 发表时间: 2006
• 期刊: Seikagaku 78
• 作者: M.;Okabe
• 通讯作者: Okabe

受精のメカニズム-イズモを中心に卵子と精子の結合に必要な因子とその異常に関連した受精異常について-

受精机制 - 以出云为中心的卵子和精子结合所必需的因素，以及与这些异常相关的受精异常 -

• 发表时间: 2006
• 期刊: 産科と婦人科 73(6)
• 作者: 井上 直和;伊川 正人;岡部 勝
• 通讯作者: 岡部 勝

遺伝子操作マウスを用いて見る卵子と精子の相互作用

使用基因工程小鼠研究卵子和精子之间的相互作用

• 发表时间: 2006
• 期刊: 生化学 78
• 作者: 井上 直和;ほか;岡部 勝
• 通讯作者: 岡部 勝

受精の膜融合における必須分子lzumoの同定

受精过程中膜融合的重要分子 lzumo 的鉴定

• 发表时间: 2006
• 期刊: Medical Science Digest 32(10)
• 作者: 井上 直和;岡部 勝
• 通讯作者: 岡部 勝

HANP1/H1T2, a Novel Histone H1-Like Protein Involved in Nuclear Formation and Sperm Fertility

• DOI: 10.1128/mcb.25.16.7107-7119.2005
• 发表时间: 2005-08
• 期刊: Molecular and Cellular Biology
• 影响因子: 5.3
• 作者: Hiromitsu Tanaka;N. Iguchi;A. Isotani;K. Kitamura;Y. Toyama;Y. Matsuoka;M. Onishi;K. Masai;M. Maekawa;K. Toshimori;M. Okabe;Y. Nishimune