Searching for the factors concerning hematopoiesis

寻找与造血相关的因素

基本信息

批准号：
10557012
负责人：
NAKANO Toru
金额：
$ 6.72万
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：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10557012/
关键词：
hematopoietic stem cells embryogenesis cell differentiaition secretory protein signal transduction

项目摘要

One of the purposes of this study is cloning the genes of the factors that have some roles in early hematopoiesis. For this purpose, we combined the in vitro hematopoietic differentiation induction method from mouse embryonic stem cells and the signal sequence trap method that is a method to select secretory proteins containing signal peptide. cDNA library was constructed from the day 5 induced mesodermal cell colonies in which hemangioblasts and hematopoietic progenitors existed. As a result, cDNA named ESOP-1 was cloned. The deduced amino acid sequence shows 160 amino acids protein. Human ESOP-1 was also cloned and shows 64% homology. ESOP-1 cDNA was highly expressed in embryos at day 7.5 of gestation. This gene was also expressed in both adult and embryonic hematopoietic system. Antibody was produced against this protein and immuno staining revealed that the protein was expressed in yolk sac, developing nervous system and adult genital organs.The other outcome is the analysis of the … More fuction of Bcl-X gene on erythropoiesis. We utilized Bcl-X null mouse embryonic stem cells (ES cells), and showed that Bcl-X is indispensable for the production of both embryonic primitive erythrocytes (EryP) and adult definitive erythrocytes (EryD) at the end of their maturation. In vivo, bcl-x null ES cells did not contribute to circulating EryD in adult chimeric mice that were produced by blastocyst microinjection of the bcl-x null ES cells. bcl-x null EryP and EryD were produced by in vitro differentiation induction of ES cells on a macrophage colony-stimulating factor deficient stromal cell line OP9, and further analysis was carried out. The emergence of immature EryP and EryD from bcl-x null ES cells was similar to that from normal ES cells. However, prominent cell death of bcl-x null EryP and EryD occurred when the cells matured. Erythropoietin is thought to prevent the apoptotic cell death of erythroid progenitor cells, while the anti-apoptotic function of bcl-x acts at the very end of maturation. Less

这项研究的目的之一是克隆在早期造血中具有某些作用的因素的基因。为此，我们合并了从小鼠胚胎干细胞中的体外造血分化诱导的方法和信号序列陷阱方法，该方法是一种选择含有信号肽的秘密蛋白质的方法。 cDNA文库是从第5天诱导的中胚层细胞菌落构建的，其中存在血管细胞和造血祖细胞。结果，克隆了名为ESOP-1的cDNA。推导的氨基酸序列显示160个氨基酸蛋白。人ESOP-1也被克隆并显示64％同源。 ESOP-1 cDNA在妊娠第7.5天在胚胎中高度表达。该基因在成人和胚胎造血系统中也表达。对这种蛋白质产生抗体，免疫染色表明该蛋白是在蛋黄囊中表达的，发展神经系统和成年生殖器官。其他结果是对促红细胞生成的Bcl-X基因的分析。我们利用了Bcl-X null小鼠胚胎干细胞（ES细胞），并表明Bcl-X对于在成熟结束时的胚胎原始红细胞（ERYP）和成年定性红细胞（ERYD）都是必不可少的。在体内，Bcl-X NULL ES细胞没有促进成年嵌合小鼠的循环红细胞，这些小鼠是由Bcl-X Null ES细胞的胚泡微注射产生的。通过在巨噬细胞刺激因子缺乏基质细胞系OP9上对ES细胞的体外分化诱导产生Bcl-X Null ERYP和ERYD，并进行了进一步的分析。来自Bcl-X NULL ES细胞的未成熟ERYP和ERYD的出现与正常ES细胞的出现相似。然而，当细胞成熟时，BCl-X Null ERYP和ERYD的明显细胞死亡发生。促红细胞生成素可以预防红细胞祖细胞的凋亡细胞死亡，而Bcl-X的抗凋亡功能在成熟的最终作用。较少的