脳における系統的インプリンティング解析を可能にする片親由来2倍体神経細胞の作製

创建单亲二倍体神经元，实现大脑中的系统印记分析

• 批准号: 17659098
• 负责人: 宮崎 和美
• 金额: $ 2.11万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Exploratory Research
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17659098/
• 关键词: インプリンティング; 脳; 神経細胞

脳におけるインプリンティングの役割を系統的に解析するために、マウス発生工学技術を駆使して片親由来2倍体神経細胞を作製することが我々の最終目標である。平成17年度に修得した基礎的なマウス発生工学技術をもとに、平成18年度は「4倍体初期胚と母親性二倍体初期胚のアグリゲーション法による母親性二倍体細胞の作製」を行った。母親性二倍体初期胚は胎児成分を構成することができるが、胎盤を形成する事ができず、正常な胎児を産出できない。これを克服するために胎盤成分を構成する細胞として4倍体初期胚を用い、この細胞と母親性二倍体初期胚とでキメラを作ることで、妊娠をE15まで継続させ母親由来2倍体神経細胞を作製することとした。ICRの正常受精卵の2分割卵を細胞融合法にて1分割卵にして培養し4倍体初期胚とする。この4倍体胚は胎盤を構成するが、胎児を構成することはできない初期胚である。一方母親性二倍体初期胚の作製にはF1ハイブリッドマウス(C57BL6xPWK)の未受精卵をエタノール処理し、卵割をおこさせる。この細胞とICR4倍体初期胚とを用いてアグリゲーション法にてキメラを作り、桑実胚に分化した時点で子宮に戻し、E15で胎児をとりだし、脳の分散培養を行い母親由来2倍体神経細胞を得ようとする実験である。上記実験を試み妊娠が継続されることを確認した。E15の時点で肉眼で胎盤、胎児を確認できたが、胎児は既に死亡しており、E15胎児脳からの神経細胞の培養には至らなかった。今後さらに条件を検討し、4倍体初期胚と母親性二倍体初期胚の4倍体初期胚と母親性二倍体初期胚のアグリゲーション法による母親由来2倍体神経細胞の回収を目指す予定である。

In order to make sure that the diploid diploid cell is the source of the diploid diploid, this is the most important part of the computer science and technology system. In the year of Pingcheng, the students who got the basic knowledge of hygiene and technology in Pingcheng in 17 and in Pingcheng in 18, in the early stage of tetraploid, the early stage of maternal diploid, the early stage of maternal diploid, the method of maternal diploid was used to test the cells of maternal diploid. In the early stage of maternal diploid, the components of the embryo were formed, the formation of the embryo, and the normal pregnancy. In the early stage of tetraploid, the embryo in the early stage of tetraploid, the embryo in the early stage of maternal diploid, the embryo in the early stage of maternal diploid, the embryo in the early stage of maternal diploid, the embryo in the early stage of maternal diploid, the embryo in the early stage of maternal diploid, the embryo in the early stage of maternal diploid, the embryo in the early stage of maternal diploid, the embryo in the early stage of maternal diploid, the embryo in the early stage of maternal diploid, the embryo in the early stage of maternal diploid, the embryo in the early stage of maternal diploid, and the mother in pregnancy. ICR "normal fertilized eggs", "2 division of eggs", "cell fusion method", "1 division of eggs", "culture" of tetraploid embryos in the early stage. The tetraploid embryos were divided into two groups: the first stage of the embryo, the second stage of the embryo, and the first stage of the embryo. The early embryos of one maternal sexual diploid were divided into two groups: unfertilized eggs (C57BL6xPWK), unfertilized eggs (F1), unfertilized eggs (Fig.1). In the early stage of ICR4 ploidy, the embryos in the early stage of embryo ploidy were used in the early stage of embryo ploidy, the embryos of mulberry embryos were differentiated at the time point of differentiation, the embryos of E15 embryos, the embryos of E15 embryos and the mother of diploid embryos were obtained by diploid culture. Please check your pregnancy and confirm your pregnancy. At the time of E15, the naked eye pregnancy, fetal confirmation, fetal death, and E15 fetal pregnancy were performed. From now on, the condition is not clear, the tetraploid embryo is the early stage of maternal diploid, the early stage of maternal diploid is the early stage of maternal diploid, the early stage of maternal diploid is the early stage of maternal diploid, the method is that the diploid diploid comes from the diploid diploid, the cell is returned to the target to indicate the predetermined embryo.

项目成果

Expression of the Snurf-Snrpn IC transcript in the oocyte and its putative role in the imprinting establishment of the mouse 7C imprinting domain.

Snurf-Snrpn IC 转录物在卵母细胞中的表达及其在小鼠 7C 印迹域印迹建立中的假定作用。

• 期刊: J.Hum.Genet. in press
• 作者: Cheng X;Kanki T;Fukuoh A;Ohgaki K;Takeya R;Aoki Y;Hamasaki N;Kang D.;Yamasaki Y;Mapendano CK
• 通讯作者: Mapendano CK