Topographic Gene Expression in Developing Brain

大脑发育中的地形基因表达

• 批准号: 6368584
• 负责人: JON S MORROW
• 金额: $ 20.44万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-20 至 2003-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6368584
• 关键词: brain injury; complementary DNA; developmental neurobiology; gene expression; genetic mapping; hypoxia neonatorum; laboratory rat; mathematical model; microarray technology; molecular biology information system; neuroanatomy; neurogenetics

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.

描述（由申请人提供）：该提案的总体目的是 证明研究脑慢性的病理生理学的可行性 通过分析基因表达模式，由大脑排序 地形，这是由于发育中的大脑损伤而发生的。我们的重点 将开发开发所需的技术和计算方法 在大规模的时间和地形上分析这些变化 平行方式，以打开载有一个新颖的探索途径 了解深度神经发育的分子基础 伴随着非常过早的出生的缺陷。我们已经建立了 记录了一个出色的动物模型（老鼠），该模型忠实地再现了许多 这种疾病的方面。我们的方法是探索 应用cDNA微阵列的检测功能，并与 高通量组织微阵列和方法的筛选功能 对原位表达曲线的敏感多路复用检测，以建立 定义与大脑特定区域有关的基因的数据库及其 对发育和潜在缺氧的反应。计算算法将是 开发的旨在帮助该数据库的分析和传播 专门的cDNA微阵列将用于其他研究人员 通过耶鲁大学的凯克实验室DNA阵列设施。这些目标将被实现 通过以下目的：1）构建cDNA微阵列至少包含8,000个 大鼠基因与发育中的大鼠神经系统中的基因表达相关； 2） 使用大鼠脑特异性cDNA阵列分析表达方式 大脑成熟过程中成千上万的基因及其对缺氧的反应； 3） 采用现代数据分析工具来定义基因层次结构或集群 参加我们的模型系统中对缺氧的反应的编排； 4）将基因表达簇映射到指定的细胞和区域 大脑利用由对照和 实验动物，通过激光捕获显微解剖和其他 方法论。总的来说，这些研究将为 调查这个毁灭性的公共卫生问题，并建立关键 未来研究其他脑疾病的方法论。