Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA.
Proc Natl Acad Sci U S A. 2010 Mar 16;107(11):5254-9. doi: 10.1073/pnas.0914114107. Epub 2010 Mar 1.
To examine the fundamental mechanisms governing neural differentiation, we analyzed the transcriptome changes that occur during the differentiation of hESCs into the neural lineage. Undifferentiated hESCs as well as cells at three stages of early neural differentiation-N1 (early initiation), N2 (neural progenitor), and N3 (early glial-like)-were analyzed using a combination of single read, paired-end read, and long read RNA sequencing. The results revealed enormous complexity in gene transcription and splicing dynamics during neural cell differentiation. We found previously unannotated transcripts and spliced isoforms specific for each stage of differentiation. Interestingly, splicing isoform diversity is highest in undifferentiated hESCs and decreases upon differentiation, a phenomenon we call isoform specialization. During neural differentiation, we observed differential expression of many types of genes, including those involved in key signaling pathways, and a large number of extracellular receptors exhibit stage-specific regulation. These results provide a valuable resource for studying neural differentiation and reveal insights into the mechanisms underlying in vitro neural differentiation of hESCs, such as neural fate specification, neural progenitor cell identity maintenance, and the transition from a predominantly neuronal state into one with increased gliogenic potential.
为了研究神经分化的基本机制，我们分析了 hESC 向神经谱系分化过程中发生的转录组变化。使用单读、配对读和长读 RNA 测序相结合的方法，分析了未分化的 hESC 以及处于早期神经分化的三个阶段（早期启动、神经祖细胞和早期胶质样）的细胞。结果显示，在神经细胞分化过程中，基因转录和剪接动态具有巨大的复杂性。我们发现了以前未注释的转录本和每个分化阶段特异性的剪接异构体。有趣的是，剪接异构体的多样性在未分化的 hESC 中最高，分化后降低，这种现象我们称之为异构体特化。在神经分化过程中，我们观察到许多类型基因的差异表达，包括参与关键信号通路的基因，并且大量细胞外受体表现出阶段特异性调节。这些结果为研究神经分化提供了有价值的资源，并揭示了 hESC 体外神经分化的机制，例如神经命运特化、神经祖细胞身份维持以及从以神经元为主的状态向具有增加的神经发生潜力的状态的转变。
