Molecular Genetic Analysis of Mammalian Neuronal differentiation

描述（由申请人提供）：神经干/祖细胞研究为理解病因和开发许多发育和行为脑疾病和脑癌的潜在治疗提供了新的途径。这项研究的核心是神经干细胞/祖细胞的两种关键状态，即自我更新状态和分化状态，是如何通过内在和外在机制在发育和成年生活中维持微妙的平衡的，因为神经干细胞/祖细胞这种稳态的缺陷会导致大脑畸形，甚至可能导致肿瘤的形成。过去十年的研究迄今为止已经发现了几十个基因，这些基因通过促进自我更新或刺激分化对这种调节很重要，然而，这一进展只代表了我们对这一基本干细胞生物学问题的理解的开始。在这项应用中，我们建议系统和全面地表征神经祖细胞中特异性表达的基因，并利用这些信息进一步确定控制神经祖细胞稳态的关键原因。我们的基本原理是，通过了解参与这一过程的相关因素，我们将更好地阐明有助于确定神经祖细胞自我更新和分化状态的机制。为此，我们开发了一种新的遗传双报告系统，可以从发育中的小鼠大脑中分离内源性神经祖细胞及其直接后代。纯化的神经祖细胞和后代将允许我们进行比较基因表达分析，以识别差异表达的基因。通过识别和表征这些差异表达的基因，我们期望发现神经祖细胞稳态的关键调节因子。这将最终有助于理解神经祖细胞中分子相互作用的平衡如何引导自我更新和分化之间的平衡，以及个体分子轴的失调如何导致大脑疾病的特定病理状态。

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.