Topographic Gene Expression in Developing Brain

大脑发育中的地形基因表达

• 批准号: 6530021
• 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/6530021
• 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）：该提案的总体目标是 证明研究脑慢性病病理生理学的可行性 通过分析基因表达模式（按大脑排序）来确定亚致死缺氧 地形，是由于发育中的大脑受伤而发生的。我们的重点 将致力于开发所需的技术和计算方法 大规模地分析这些时间和地形上的变化 平行的方式，从而开辟新的探索途径，影响 了解深刻的神经发育的分子基础 伴随早产的缺陷。我们已经建立并 记录了一个优秀的动物模型（大鼠），它忠实地再现了许多 这种疾病的各个方面。我们的方法是探索可行性 应用 cDNA 微阵列的检测能力，并结合 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 一个数据库，定义与大脑特定区域相关的基因及其 对发育和亚致死缺氧的反应。计算算法将是 开发用于帮助分析和传播该数据库，以及 专门的 cDNA 微阵列将可供其他研究人员使用 通过耶鲁大学凯克实验室 DNA 阵列设施。这些目标将会实现 通过以下目标： 1) 构建包含至少 8,000 个 cDNA 微阵列 与发育中的大鼠神经系统基因表达相关的大鼠基因； 2） 使用大鼠脑特异性 cDNA 阵列分析 大脑成熟过程中的数千个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聚类 参与我们模型系统中对缺氧反应的协调； 4) 将基因表达簇映射到指定的细胞和区域 大脑利用由对照和构建的高通量组织微阵列 实验动物，通过激光捕获显微切割和其他方法进行验证 方法论。总的来说，这些研究将为 调查这一毁灭性的公共卫生问题，并建立关键 未来研究其他脑部疾病的方法。