Functional Genomic Studies of Neuronal Differentiation

脊椎动物神经系统内神经元的精确分化由一个复合体来确定 转录因子相互作用的遗传程序，以及通过 环境线索。尽管许多特定的转录因子和信号通路的作用 关于神经元分化的研究，我们对其具体机制的理解还不够深入。 这还远远没有完成。这一建议使用了几个哺乳动物基因组的基因组序列信息， 脊椎动物神经系统发育自始至终高度保守的基本观察 进化和功能基因组学的最新进展，以提出高通量功能基因组学 神经元分化的分析。这个提议要检验的假设是进化论 在神经元分化过程中诱导的基因周围的保守序列对 神经元诱导的编排。在第一个特定目标中，将诱导P19胚胎性癌细胞 为了进行神经元分化和微阵列杂交研究，将进行基因鉴定 受转录调控的基因。在第二个具体目标中，进化保守的非精灵 小鼠基因组的序列(CNG)，最近被提出代表顺式-DNA序列簇。 调控序列，将使用高通量微阵列分析转录调控 P19细胞的转染法。在第三个特定目标中，蛋白激酶和转录的作用 调控这些顺式作用序列的因素将被描述。这些研究的最终结果是 将详细了解神经诱导基因的重要顺式调控序列和 特定信号转导通路在其转录激活调控中的作用。结果是 这些研究将对未来治疗神经退行性变的方法具有重要意义 包括阿尔茨海默氏症和帕金森氏症在内的疾病。

The precise differentiation of neurons within the vertebrate nervous system is specified by a complex genetic program of transcription factor interactions, as well as modulation of these interactions by environmental cues. Although the roles of many specific transcription factors and signaling pathways have been characterized in neuronal differentiation, our understanding of the detailed mechanisms involved is still far from complete. This proposal employs genomic sequence information of several mammalian genomes, the fundamental observation that vertebrate nervous system development is highly conserved throughout evolution, and recent developments in functional genomics to propose a high-throughput functional genomic analysis of neuronal differentiation. The hypothesis to be tested in this proposal is that evolutionary conserved sequences surrounding genes induced during neuronal differentiation are important to the orchestration of neuronal induction. In the first specific aim,P19 embyronic carcinoma cells will be induced to undergo neuronal differentiation and microarray hybridization studies will be carried out to identify genes that are transcriptionally regulated. In the second specific aim, evolutionarily conserved non-genie sequences (CNGs) of the mouse genome, that have recently been proposed to represent clusters of cis- regulatory sequences, will be analyzed for transcriptional regulation using a high-throughput microarray transfection method in P19 cells. In the third specific aim, the role of protein kinases and transcription factors in the regulation of these cis- acting sequences will be characterized. The end result of these studies will be a detailed understanding of important cis-regulatory sequences of neuronally induced genes and the role of specific signal transduction pathways in the modulation of their transcriptional activation. The results of these studies will be important for future therapeutic approaches to treatment of neurodegenerative diseases including Alzheimer's and Parkinson's diseases.