Study of implantation using molecular biological means using xeno-chimeric embryos.

使用异种嵌合胚胎利用分子生物学手段研究植入。

• 批准号: 14380383
• 负责人: OKABE Masaru
• 金额: $ 10.24万
• 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: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14380383/
• 关键词: green mouse; green rat; red mouse; implantation; chimera; MAPC cells; 異種胚キメラ; ジフテリアトキシン; Creリコンビナーゼ; トランスジェニックマウス; loxP; 骨髄細胞; 幹細胞; GFP; 異種移植; ラット; マウス; 妊娠

We investigated the mechanism of implantation using molecular biological techniques and transgenic animals. We have established "green rat" lines together with "red mouse" lines. The combination of these zeno-clilmeric embryos allows us to differentiate the contribution of cells from which species the cells are derived from.In order to have a sufficient source from rat embryos, we have established multipotent stem cells from "green rat" bone marrow cells (MAPC). We also succeeded to establish the stem cells of trophectoderm origin.In the middle of producing chimeric mice, We produced XX7XY chimeras by using embryos whose X chromosomes were tagged with EGFP (X^*), making the fluorescent green female (XX^*) germ cells easily distinguishable from their nonfluorescent male (XY) counterparts. Taking advantage of tagging with EGFP, the XX^* "prospermatogonia" were isolated from the testes, and the status of their genomic imprinting was examined. It was shown that these XX cells underwent a paternal imprinting, despite their chromosomal constitution. As previously indicated in sex-reversal XXsxr testes, we also found a few green XX^* germ cells developed as "eggs" within the seminiferous tubules of XX^*7XY chimeric testes. These cells were indistinguishable from XX^* prospermatogonia at birth but resumed oogenesis in a testicular environment. The biological nature of the "testicular eggs" was examined by recovering the eggs from chimeric testes.The testicular eggs not only formed an egg-specific structure, the zona pellucida, but also were able to fuse with sperm. The collected testicular eggs were indicated to undergo maternal imprinting, despite the testicular environment. The genomic imprinting did not always follow the environmental conditions of where the germ cells resided ; rather, it was defined by the sex that was chosen by the germ cells at early embryonic stage.

我们使用分子生物学技术和转基因动物研究了植入机制。我们已经与“红色鼠标”线建立了“绿老鼠”线。这些Zeno-Clilmeric胚胎的组合使我们能够区分细胞从中得出的细胞的贡献。为了从大鼠胚胎中获得足够的来源，我们已经建立了多能干细胞与“绿老鼠”骨髓细胞（MAPC）。我们还成功地建立了滋养剂的干细胞。在产生嵌合小鼠的中间，我们通过使用其X染色体标记为EGFP（X^*）的胚胎产生XX7XY嵌合体，从而使荧光绿色雌性（XX^*）易于区分其非荧光男性（Xy）counterparts。利用与EGFP进行标记，XX^*“ Prospermatogonia”是从睾丸中分离出来的，并检查了其基因组印记的状态。结果表明，尽管具有染色体构成，但这些XX细胞还是经历了父亲的印记。如前所述，在性逆转XXSXR睾丸中，我们还发现了XX^* 7x 7 Xy嵌合睾丸的绿化小管中的一些绿色XX^*生殖细胞以“卵”形成。这些细胞在出生时与xx^* Prospermatogonia没有区别，但在睾丸环境中恢复了卵子。通过从嵌合睾丸中回收卵，检查了“睾丸卵”的生物学性质。睾丸卵不仅形成了卵特异性结构，即Zona Pellucida，而且还可以与精子融合。尽管有睾丸环境，但收集的睾丸卵被指出会经历母体的印记。基因组印迹并不总是遵循生殖细胞所在的环境条件。相反，它是由生殖细胞在早期胚胎阶段选择的性别所定义的。

项目成果

Small interfering RNA and gene silencing in transgenic mice and rats

• DOI: 10.1016/s0014-5793(02)03680-3
• 发表时间: 2002-12-04
• 期刊: FEBS LETTERS
• 影响因子: 3.5
• 作者: Hasuwa, H;Kaseda, K;Okabe, M
• 通讯作者: Okabe, M

Nomura M et al.: "Mechanism of host cell protection from complement in murine cytomegalovirus(CMV) infection : identification of a CMV-responsive element in the CD46 promoter region"Eur J Immunol. 32(10). 2954-2964 (2002)

Nomura M 等人：“鼠巨细胞病毒 (CMV) 感染中宿主细胞免受补体侵害的机制：CD46 启动子区域中 CMV 响应元件的鉴定”Eur J Nutrition。

Genomic imprinting of XX spermatogonia and XX oocytes recovered from XX{leftrightarrow}XY chimeric testes

从 XX{leftrightarrow}XY 嵌合睾丸中回收的 XX 精原细胞和 XX 卵母细胞的基因组印记

• 发表时间: 2005
• 期刊: Proc Natl Acad Sci USA 102(11)
• 作者: A.Isotani et al.

Lineage-specific cell disruption in living mice by Cre-mediated expression of diphtheria toxin A chain

• DOI: 10.1016/j.bbrc.2004.06.139
• 发表时间: 2004-08-20
• 期刊: BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
• 影响因子: 3.1
• 作者: Matsumura, H;Hasuwa, H;Okabe, M
• 通讯作者: Okabe, M

Migishima F. et al.: "Full reconstitution of hematopoietic system by murine umbilical cord blood"Transplantation. 75(1). 1820-1826 (2003)

Migishima F.等人：“通过鼠脐带血完全重建造血系统”移植。