Microvascular endothelial cell heterogeneity in the central nervous system

中枢神经系统微血管内皮细胞异质性

• 批准号: 7342513
• 负责人: Joel S Pachter
• 金额: $ 16.19万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-01-22 至 2009-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7342513
• 关键词: Address; Affect; Area; Attention; Behavior; Biochemistry; Biological; Blood; Blood - brain barrier anatomy; Blood Vessels; Blood capillaries; Brain; Capillary Endothelial Cell; Cerebral cortex; Cerebrum; Classification; Condition; Coupling; DNA; DNA Microarray Chip; DNA Microarray format; Data; Depth; Development; Disease; Drug Formulations; Endothelial Cells; Evaluation; Exploratory/Developmental Grant; Gene Expression; Gene Expression Profiling; Genes; Goals; Grant; Health; Heterogeneity; Homeostasis; In Situ; Inflammatory; Investigation; Lead; Light; Location; Microarray Analysis; Modeling; Molecular; Molecular Profiling; Nature; Neuraxis; Neurons; Parietal Lobe; Pattern; Peripheral; Permeability; Physiological; Population; Process; RNA; Rate; Reporting; Reproducibility; Research; Risk; Sampling; Source; Spinal Cord; Staging; Support Contracts; System; Techniques; Testing; Tissues; Transcript; Trees; United States National Institutes of Health; arteriole; base; capillary; design; hemodynamics; laser capture microdissection; neurovascular unit; novel; tissue culture; tool; vascular bed; venule

The issue of microvascular endothelial heterogeneity greatly impacts vascular function in both health and disease, and potentially severly complicates the use of tissue culture-based models to study endothelial behavior. Yet, despite this topic having been explored in considerable depth in peripheral vascular beds, surprisingly little attention has yet been focused on the microvascular axis of central nervous system (CNS). Unrelated reports describing particular genes have, nonetheless, provocatively suggested that a diversity in endothelial gene expression may exist at the segmental level (i.e., between arterioles, capillaries and venules) as well as the regional level of the CNS, and approximate or surpass that found in the periphery. Given the preeminance of the microvascular network in forming both the blood-brain barrier and blood- spinal cord barrier, and the significant microvascular involvement in inflammatory, infectious, degenerative and traumatic conditions of the CNS, it is critical that there be a systematic and detailed evaluation of this cliinically important issue. It is of further importance that such analysis be performed on microvascular tissue in situ," as endothelial cell gene expression is exquisitely prone to environmental modulation. Accordingly, we propose the following Specific Aims: 1) To validate and optimize the approach of coupling laser capture microdissection (LCM) of brain microvascular endothelial cells, with global gene expression profiling byDMA microarray analysis. This will include demonstrating feasibility, by maximizing the percentage of transcripts detected (P call rate) in capillaries only, and establishing reproducibility, by identifying both technical and biological sources of variance in microarray data; and 2) To use the LCM/microarray approach to compare the global gene expression profiles of endothelial cells from capillaries, venules and arterioles. The R21 format chosen is specifically designed for projects that are exploratory and may involve considerable risk, but lead to the development of novel techniques that could have major impact on a field of research. In this regard, results obtained here will set the stage for evaluating regional gene expression by the neurovascular unit (i.e,endothelial cells and intimately associated neural cells) along the CNS microvascular tree in both health and disease. In turn, these studies will enable formulation of more precise endothelial models to study the molecular basis of physiological and pathophysiological processes of the cerebral microvasculature.

微血管内皮异质性的问题极大地影响了健康和 疾病，并有可能严重复杂组织基于组织培养的模型研究内皮 行为。然而，尽管这个话题在周围血管床上进行了相当深的探索，但 令人惊讶的是，几乎没有关注中枢神经系统（CNS）的微血管轴。 尽管如此，描述特定基因的不相关报告挑衅地表明，多样性的多样性 内皮基因表达可能存在于节段水平（即，在小动脉，毛细血管和之间 静脉）以及中枢神经系统的区域水平，以及在周围中发现的近似或超越。 考虑到微血管网络在形成血脑屏障和血液 - 脊髓屏障以及显着的微血管参与炎症，传染性，退化性 和中枢神经系统的创伤状况，至关重要的是对此进行系统的详细评估 克里率重要的问题。在微血管组织上进行此类分析是更重要的 原位，“由于内皮细胞基因表达非常容易受到环境调节的限制。因此， 我们提出以下特定目的：1）验证和优化耦合激光捕获的方法 脑微血管内皮细胞的微分解（LCM），全球基因表达分析bydma 微阵列分析。这将包括通过最大化笔录的百分比来证明可行性 仅在毛细管中检测到（p呼叫率），并通过确定技术和 微阵列数据中差异的生物学来源； 2）使用LCM/微阵列方法比较 毛细血管，静脉和小动物的内皮细胞的全球基因表达谱。 R21 选择的格式专门为探索性的项目而设计，可能涉及很大的风险，但是 导致新技术的发展，这些技术可能会对研究领域产生重大影响。在这个 注意，此处获得的结果将为评估神经血管的区域基因表达奠定阶段 沿着CNS微血管树的单位（即内皮细胞和密切相关的神经细胞） 健康与疾病。反过来，这些研究将使更精确的内皮模型制定能够研究 脑微脉管系统的生理和病理生理过程的分子基础。