Proteomic identification of differentially expressed genes during differentiation of cynomolgus monkey (Macaca fascicularis) embryonic stem cells to astrocyte progenitor cells in vitro.
Proteomic identification of differentially expressed genes during differentiation of cynomolgus monkey (Macaca fascicularis) embryonic stem cells to astrocyte progenitor cells in vitro.
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食蟹猴胚胎干细胞体外分化为星形胶质细胞祖细胞过程中差异表达基因的蛋白质组学鉴定。
DOI：
10.1016/j.bbapap.2012.12.002
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
赤間邦子
中科院分区：
文献类型：
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作者：
Toda T;Nakamura M;Morisawa H;Hirota M;Nishigaki R;Yoshimi Y.;高木大樹;赤間邦子
了解星形胶质细胞的发生对于神经系统疾病的治疗是有价值的，因为星形胶质细胞为神经元提供结构、代谢和防御支持，并主动调节神经元。然而，有有限的信息与灵长类动物ES细胞分化为星形胶质细胞的分子事件。因此，我们研究了早期星形胶质细胞发生中的差异表达的蛋白质，从食蟹猴ES细胞（CMK6细胞系）到星形胶质细胞祖细胞（AstP）细胞通过形成原始神经干球（第4天），成熟神经干球（NSS），神经干细胞（NS）在体外，使用二维凝胶电泳（2-DE）和液相色谱-串联质谱（LC-MS-MS）。我们确定了66个差异表达的蛋白质参与这五个分化阶段。结合蛋白质印迹、RT-PCR和与鉴定的蛋白质相关的代谢途径的搜索的结果，这些结果表明，从ES细胞到第4天和NSS细胞，分化阶段凋亡相关蛋白如半胱天冬酶可能与哪些分化阶段有关;从NSS细胞到NS细胞，磷酸化CRMP2进一步上调，但CRABP 1下调，在分化阶段，可能准备了大量的轴突导向蛋白，用于生长锥的发育、轴突吸引和排斥;非磷酸化CRMP 2下调，但CRABP 1从NS细胞到AstP细胞重新上调，在分化阶段可能伴随着与粘着斑有关的肌动蛋白细胞骨架的重组。这些结果为猴早期星形胶质细胞发生的分子基础提供了新的认识。
Understanding astrocytogenesis is valuable for the treatment of nervous system disorders, as astrocytes provide structural, metabolic and defense support to neurons, and regulate neurons actively. However, there is limited information about the molecular events associated with the differentiation from primate ES cells to astrocytes. We therefore investigated the differentially expressed proteins in early astrocytogenesis, from cynomolgus monkey ES cells (CMK6 cell line) into astrocyte progenitor (AstP) cells via the formation of primitive neural stem spheres (Day 4), mature neural stem spheres (NSS), and neural stem (NS) cells in vitro, using two-dimensional gel electrophoresis (2-DE) and liquid chromatography-tandem mass spectrometry (LC-MS-MS). We identified 66 differentially expressed proteins involved in these five differentiation stages. Together with the results of Western blotting, RT-PCR, and a search of metabolic pathways related to the identified proteins, these results indicated that collapsin response mediator protein 2 (CRMP2), its phosphorylated forms, and cellular retinoic acid binding protein 1 (CRABP1) were upregulated from ES cells to Day 4 and NSS cells, to which differentiation stages apoptosis-associated proteins such as caspases were possibly related; Phosphorylated CRMP2s were further upregulated but CRABP1 was downregulated from NSS cells to NS cells, during which differentiation stage considerable axon guidance proteins for development of growth cones, axon attraction, and repulsion were possibly readied; Nonphosphorylated CRMP2 was downregulated but CRABP1 was re-upregulated from NS cells to AstP cells, in which differentiation stage reorganization of actin cytoskeleton linked to focal adhesion was possibly accompanied. These results provide insight into the molecular basis of early astrocytogenesis in monkey.