The Role of NMD in Cortical Neural Progenitor Cells

项目总结 NMD关键因子Upf2、Upf3a、Upf3b和Smg6的突变， 富含各种神经发育疾病。除了通过降级来确保成绩单质量 带有过早终止密码子的异常转录，NMD调节选择性mRNAs的稳定性以进行微调 文字记录的丰富性。NMD是否以及如何影响大脑发育仍然难以捉摸。我们的长期合作 目的是了解NMD调节在复杂而动态的过程中的功能作用 神经发生及其错误调控如何导致神经发育障碍。我们确定了 选择性基因消融UPf2和体内NMD对神经发育的要求 操纵其他NMD因素。我们的初步数据显示，UPF2在神经干细胞和 祖细胞会导致小头畸形。UPF2缺失特异性地影响细胞周期和谱系进展 放射状胶质细胞(RGC)是发育中的新皮质中的主要神经前体细胞。我们将结合切割 边缘分子细胞核基因组学方法、小鼠遗传学和发育神经生物学 NMD调控神经发生的机制。我们提出了三个独立和相互关联的目标，以 调查小头畸形症表型潜在的可能变量。在目标1中，我们将确定细胞 NMD基因敲除小鼠视网膜神经节细胞周期行为的定性和定量研究 监管机制。在目标2中，我们将确定视网膜节细胞的谱系进化以及由此产生的 NMD基因敲除小鼠每单位时间的神经元输出。通过表征这些分子细胞缺陷，我们 也旨在为RGC谱系转变的转录调控提供机械性见解。NMD可能 独立于细胞周期控制或作为影响细胞周期的结果来调节细胞命运。在……里面 目标3，我们将测试这两个假设，并利用我们的结果重新检查细胞之间的关系 周期和细胞命运。这些研究的成功完成将提供对如何选择的基本见解 MRNA的稳定性是哺乳动物大脑中高度调控的皮质神经发生过程的基础。这个 拟议中的研究还将阐明一些基本问题，如细胞周期、细胞命运、 以及它们在神经发育过程中的关系。

PROJECT SUMMARY Mutations in key factors of nonsense-mediated mRNA decay (NMD), including Upf2, Upf3a, Upf3b, and Smg6, are enriched in various neurodevelopmental diseases. In additional to ensuring transcript quality by degrading aberrant transcripts with a premature stop codon, NMD modulates stability of selective mRNAs to fine-tune transcript abundance. Whether and how NMD influences brain development remains elusive. Our long-term objective is to understand the functional role of NMD regulation for the complicated and dynamic process of neurogenesis and how its mis-regulation leads to neurodevelopmental disorders. We determine the requirement of NMD for neural development through selective genetic ablation of Upf2 and in vivo manipulation of other NMD factors. Our preliminary data show that deletion of UPF2 in neural stem and progenitors results in microcephaly. UPF2 loss specifically affects the cell cycle and lineage progression of radial glia cells (RGCs), the major neural progenitor cells in the developing neocortex. We will combine cutting edge molecular cellular ribogenomics approaches, mouse genetics, and developmental neurobiology to dissect the mechanisms of NMD regulating neurogenesis. We propose three independent and interrelated aims to investigate possible variables underlying the microcephaly phenotype. In Aim 1, we will determine the cell cycle behaviors of RGCs in NMD knockout mice qualitatively and quantitatively and unveil the underlying regulatory mechanisms. In Aim 2, we will determine the lineage progression of RGCs and the resulting neuronal outputs per time unit in NMD knockout mice. By characterizing these molecular cellular defects, we also aim to provide mechanistic insights to transcriptomic regulation of RGC’s lineage transitions. NMD may regulate cell fates either independent of cell cycle controls or as the consequence of affecting the cell cycle. In Aim 3, we will test these two hypotheses and leverage our results to reexamine the relationship between cell cycle and cell fate. Successful completion of these studies will provide fundamental insights into how selective mRNA stability underlies the highly regulated cortical neurogenesis process in the mammalian brain. The proposed studies will also shed light on some fundamental questions about the control of cell cycle, cell fate, and their relationship during neural development.