Expression data from mNSc after 48 hour of treatment with CD95L-T4. Mus musculus

In neural stem cells, stimulation of the “death receptor” CD95 does not trigger apoptosis but resulted in increased stem cell survival and neuronal specification via activation of the Src /PI3K /AKT/mTOR signalling pathway. To further characterize CD95-dependent neural stem cell survival and differentiation we used conventional gene expression profiling combined with translation state array analysis. Mouse neural stem cells grown in neurosphere cultures were stimulated with a trimerized CD95L construct (CD95L-T4) and total as well as polysomal bound RNA was isolated 48 hours after stimulation and analysed by microarrays. CD95L-T4 treatment induced a global increase in ribosome-bound mRNA and protein translation as well as changes on genes involved in neurogenesis, protein synthesis and transcription factors. Overall design: Mouse neural stem cells grown in neurosphere cultures were stimulated with a trimerized CD95L construct (CD95L-T4) and total as well as polysomal bound RNA was isolated 48 hours after stimulation and hybridized on affymetrix microarrays.

在神经干细胞中，刺激死亡受体（death receptor）CD95并不会引发细胞凋亡（apoptosis），而是通过激活Src/PI3K/AKT/mTOR信号通路，提升干细胞存活率并促进神经元特化（neuronal specification）。为进一步阐明CD95依赖的神经干细胞存活与分化过程，我们采用常规基因表达谱分析（gene expression profiling）结合翻译状态阵列分析（translation state array analysis）。对在神经球培养（neurosphere cultures）体系中培养的小鼠神经干细胞施加三聚化CD95L构建体（trimerized CD95L construct，CD95L-T4）刺激后，于刺激后48小时分离总RNA及多聚核糖体结合RNA（polysomal bound RNA），并通过微阵列（microarrays）开展分析。CD95L-T4处理可全局上调核糖体结合mRNA水平与蛋白质翻译效率，并改变神经发生（neurogenesis）、蛋白质合成及转录因子（transcription factors）相关基因的表达。实验设计概述：对在神经球培养体系中培养的小鼠神经干细胞施加三聚化CD95L构建体（CD95L-T4）刺激后，于刺激后48小时分离总RNA及多聚核糖体结合RNA，并将其与Affymetrix微阵列芯片（Affymetrix microarrays）进行杂交。