Molecular Genetic Analysis of Mammalian Neuronal differentiation

描述(由申请人提供)：神经干细胞/祖细胞研究为了解病因和开发许多发育和行为脑部疾病和脑癌的潜在治疗方法提供了新的途径。这项研究的中心是干细胞/祖细胞的两个关键状态-自我更新状态和分化状态-之间的微妙平衡是如何由发育和成年生活中的内在和外在机制维持的，因为神经干细胞/祖细胞的这种稳态缺陷会导致大脑畸形，甚至可能导致肿瘤的形成。到目前为止，过去十年的研究已经发现了数十个基因，它们通过促进自我更新或刺激分化而对这种调控起到重要作用，然而，这一进展只是我们对这一基本干细胞生物学问题理解的开始。在这一应用中，我们建议系统和全面地表征在神经前体细胞中特异表达的基因，并利用这些信息进一步识别对控制神经前体细胞内稳态至关重要的原因因素。我们的理论基础是，通过了解这一过程中涉及的相关因素，我们将更好地阐明有助于确定神经前体细胞自我更新和分化状态的机制。为此，我们开发了一种新型的基因双报告系统，可以从发育中的小鼠脑中分离内源性神经前体细胞及其直接后代。纯化的神经前体细胞和后代将允许我们进行比较基因表达分析，以确定差异表达的基因。通过鉴定和鉴定这些差异表达的基因，我们期望发现神经前体细胞稳态的关键调节因子。这最终将有助于理解神经前体细胞中分子相互作用的平衡是什么以及如何指导自我更新和分化之间的平衡，以及单个分子轴的调节失调如何导致大脑疾病的特定病理状况。

DESCRIPTION (provided by applicant): Neural stem/progenitor cell study offers novel avenues for understanding the etiology and for developing potential treatment of many developmental and behavioral brain disorders and brain cancers. At the center of the study is how a delicate balance between the two key states of stem/progenitor cells, the self-renewal state and differentiation state, are maintained by intrinsic and extrinsic mechanisms in development and in adult life, because a defect in this homeostasis of neural stem/progenitor cells causes malformation of the brain and may even lead to tumor formation. Research over the past decade has thus far uncovered dozens of genes that are important for this regulation by either promoting self-renewal or stimulating differentiation, however, this progress represents only the beginning of our understanding on this fundamental stem cell biology issue. In this application, we propose to systematically and comprehensively characterize genes that are specifically expressed in neural progenitor cells and use this information to further identify causal factors crucial for the control of neural progenitor homeostasis. Our rationale is that by knowing the relevant factors involved in this process, we will be in a better position to elucidate the mechanisms that help specify the self-renewal and differentiation state of neural progenitor cells. To this end, we have developed a novel genetic two reporter system that enables isolation of endogenous neural progenitor cells and their direct progeny from the developing mouse brains. The purified neural progenitor cells and progeny will allow us to perform comparative gene expression analyses to identify differentially expressed genes. By identifying and characterizing these differentially expressed genes, we expect to uncover key regulators of neural progenitor homeostasis. This will ultimately help understand what and how equilibrium of molecular interactions in neural progenitor cells guides the balance between self-renewal and differentiation and how dysregulation in individual molecular axis may lead to a particular pathological condition in brain disorders.