Differential Requirement for Nucleostemin in Embryonic Stem Cell and Neural Stem Cell Viability
Differential Requirement for Nucleostemin in Embryonic Stem Cell and Neural Stem Cell Viability
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DOI：
10.1002/stem.44
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发表时间：
2009-01-01
期刊：
影响因子：
5.2
通讯作者：
Okuda, Akihiko
中科院分区：
文献类型：
--
作者：
Nomura, Jun;Maruyama, Masayoshi;Okuda, Akihiko
干细胞具有惊人的自我更新能力和产生多种细胞类型的能力。核干素是一种富含在多种干细胞中的蛋白质。在小鼠体内定向缺失核干素会导致着床阶段的发育停滞，这表明核干素对早期胚胎发育至关重要。然而，核干蛋白在小鼠早期胚胎中作用的分子基础仍然很不清楚，由于核干素缺失动物在早期胚胎中的致死性，核干素在组织干细胞中的作用还没有被基因打靶分析所证实。为了解决这些问题，我们在缺失的胚胎干细胞中产生了可诱导的核干细胞，其中核干细胞蛋白的两个等位基因都被打乱，但在四环素反应转录激活剂的控制下，核干细胞蛋白基因被引入到rosa26基因座中。我们发现，在ES细胞和ES细胞来源的神经干细胞/祖细胞中，核干素的丢失都会导致细胞增殖减少和凋亡增加。ES细胞的细胞活力下降比神经干细胞/祖细胞严重得多，这种影响至少部分是通过增加ES细胞中P53和/或激活的caspase-3的诱导和积累而不是神经干细胞/祖细胞的。干细胞2009；27：1066-1076
Stem cells have the remarkable ability to self-renew and to generate multiple cell types. Nucleostemin is one of proteins that are enriched in many types of stem cells. Targeted deletion of nucleostemin in the mouse results in developmental arrest at the implantation stage, indicating that nucleostemin is crucial for early embryogenesis. However, the molecular basis of nucleostemin function in early mouse embryos remains largely unknown, and the role of nucleostemin in tissue stem cells has not been examined by gene targeting analyses due to the early embryonic lethality of nucleostemin null animals. To address these questions, we generated inducible nucleostemin null embryonic stem (ES) cells in which both alleles of nucleostemin are disrupted, but nucleostemin cDNA under the control of a tetracycline-responsive transcriptional activator is introduced into the Rosa26 locus. We show that loss of nucleostemin results in reduced cell proliferation and increased apoptosis in both ES cells and ES cell-derived neural stem/progenitor cells. The reduction in cell viability is much more profound in ES cells than in neural stem/progenitor cells, an effect that is mediated at least in part by increased induction and accumulation of p53 and/or activated caspase-3 in ES cells than in neural stem/progenitor cells. STEM CELLS 2009;27:1066-1076