Topographic Gene Expression in Developing Brain

描述(由申请人提供)：本提案的总体目标是 论证研究慢性脑病病理生理学的可行性 通过基因表达模式分析亚致死性低氧，按大脑分类 地形图，是对发育中的大脑损伤做出反应而产生的。我们的重点是 将致力于开发所需的技术和计算方法 大规模地从时间和地形上分析这些变化 平行的方式，从而开辟了一条新的探索道路 对深层神经发育的分子基础的认识 伴随着早产而来的缺陷。我们已经建立了 记录了一个非常好的动物模型(大鼠)，它忠实地复制了许多 这种紊乱的各个方面。我们的做法是探索 应用cDNA微阵列的检测能力，与 高通量组织微阵列的筛选能力和方法 原位表达谱的灵敏多重检测，以建立 定义与大脑特定区域相关的基因的数据库，以及它们的 对发育和亚致命缺氧的反应。计算算法将是 为协助分析和传播该数据库而开发，以及 专门的cdna微阵列将提供给其他研究人员。 通过耶鲁大学凯克实验室的DNA阵列设备。这些目标将会实现 通过以下目标：1)构建至少包含8,000个DNA的微阵列 大鼠神经系统发育过程中与基因表达相关的基因；2) 利用大鼠脑内特异的基因芯片分析其表达模式 大脑成熟过程中的数千个基因及其对低氧的反应；3) 使用现代数据分析工具定义基因层次结构或集群 在我们的模型系统中参与对低氧反应的协调； 4)将基因表达簇映射到指定的细胞和区域 利用高通量组织微阵列构建的大脑 实验动物，通过激光捕获显微解剖和其他方法验证 方法论。总体而言，这些研究将开辟全新的途径 调查这一毁灭性的公共卫生问题，并建立关键 未来对其他大脑疾病的研究方法。

DESCRIPTION (provided by applicant): The overall aim of this proposal is to demonstrate the feasibility of studying the pathophysiology of cerebral chronic sublethal hypoxia by profiling gene expression patterns, sorted by brain topography, that occur in response to injury in the developing brain. Our focus will be on developing the technical and computational approaches needed to analyze these changes both temporally and topographically in a massively parallel way, so as to open novel avenues of exploration that bear on an understanding of the molecular basis of the profound neurodevelopmental deficiencies that accompany very premature birth. We have established and documented an excellent animal model (rat) that faithfully reproduces many aspects of this disorder. Our approach is to explore the feasibility of applying the detection capabilities of cDNA microarrays, coupled with the screening capabilities of high-throughput tissue microarrays and methods for the sensitive multiplex detection of in-situ expression profiles, to establish a database defining genes relevant to specific regions of the brain, and their response to development and sublethal hypoxia. Computational algorithms will be developed to aid in the analysis and dissemination of this database, and specialized cDNA microarrays will be made available for other investigators through Yale's Keck Laboratory DNA array facility. These goals will be achieved by the following aims: 1) Construct cDNA microarrays comprising at least 8,000 rat genes relevant to gene expression in the developing rat nervous system; 2) Use the rat brain specific cDNA arrays to analyze the patterns of expression of thousands of genes during brain maturation and its response to hypoxia; 3) Employ modern data analysis tools to define gene hierarchies or clusters that participate in the orchestration of responses to hypoxia in our model system; and 4) Map the gene expression clusters to specified cells and regions in the brain utilizing high-throughput tissue microarrays constructed from control and experimental animals, validated by laser capture microdissection and other methodologies. Collectively, these studies will open entirely new avenues for investigation of this devastating public health problem, and establish key methodologies for future investigations of other brain disorders.