Effects of VEGF on Brain Tissue in vitro

VEGF对体外脑组织的影响

• 批准号: 6540181
• 负责人: JEFFREY M ROSENSTEIN
• 金额: $ 30.4万
• 依托单位: GEORGE WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6540181
• 关键词: Animalia; angiogenesis; biological signal transduction; blood vessels; cell growth regulation; cell proliferation; central nervous system; cerebrovascular system; confocal scanning microscopy; cytokine; developmental neurobiology; glia; glucose transporter; growth factor receptors; immunoaffinity chromatography; in situ hybridization; microdialysis; neurogenesis; neuropil; organ culture; protein structure function; time resolved data; vascular endothelial growth factors; western blottings

DESCRIPTION (provided by applicant): Vascular endothelial growth factor (VEGF) is a secreted growth factor that plays an important regulatory role in vascular development. VEGF is a significant factor in angiogenesis in the periphery and in tumors and it has already been used clinically for peripheral vascular deficiencies. The proposed experiments will advance our understanding of VEGF and its receptors with respect to brain development and plasticity and may determine if the growth factor plays multiple roles in the CNS. Presently, there exists a paucity of work regarding its direct cellular actions on CNS tissues. The broad long-term goals of this research program are to understand the cellular and molecular interactions of VEGF with CNS tissue with a dual in vitro approach. Experiments use a serum-free organotypic explant system to determine VEGF effects on neuropil at different perinatal developmental stages as well as primary culture assays of neurons, astroglia and isolated CNS vessels. Preliminary data indicate that applied VEGF causes significant angiogenesis in fetal and adult CNS. Unexpectedly, VEGF application also produces significant astroglial proliferation mediated by its flt-1 receptor and enhanced neuronal expression of developmentally regulated proteins and as well as significant neurite outgrowth mediated by its flk-1 receptor. S.A. 1 will examine the angiogenic capacity of applied VEGF to CNS explants as well as the regulation of VEGF receptors and the integrity of the blood-brain barrier. S.A. 2 will test the hypothesis that VEGF acts as a mitogen for astroglia and is a growth factor for CNS neurons in organotypic explants. VEGF receptor and neuronal and astroglial markers will be examined with confocal microscopy, in situ hybridization and quantified by immunoblotting and immunoaffinity chromatography. An innovative technology utilizes microculture chambers to permit real-time microdialysis to determine if VEGF induces production of other cytokines/growth factors. Application of non-specific proteins, anti-sense oligodeoxynucleotides and receptor binding proteins will determine VEGF/receptor specificity in the system. We will also initiate experiments for analysis of receptor signal transduction pathways. S.A. 3 will test comparable parameters in primary cultures in order to verify VEGF's direct effects on CNS elements, absent potential indirect actions between cell types. As an angiogenic factor VEGF could have potential use in cerebrovascular disorders but may also have trophic effects upon astroglia and neurons. The results can lay important cell biological groundwork for understanding how brain tissue might respond to VEGF-based therapy.

描述（由申请人提供）：血管内皮生长因子（VEGF） 是一个分泌的生长因子，在血管中起重要的调节作用 发展。 VEGF是周围血管生成的重要因素， 在肿瘤中，已经在临床上用于周围血管 缺陷。提出的实验将提高我们对VEGF的理解 及其受体在大脑发育和可塑性方面，可能 确定生长因子是否在中枢神经系统中起多重作用。目前， 关于其直接细胞对CNS的直接细胞作用存在很少的工作 组织。该研究计划的长期目标是了解 VEGF与CNS组织的细胞和分子相互作用与双重 体外方法。实验使用无血清器官型外植体系统 确定VEGF对不同围产期发育阶段神经肽的影响 以及神经元，星形胶质细胞和孤立的CNS的原发性培养测定法 船只。初步数据表明，应用VEGF会引起显着 胎儿和成人中枢神经系统的血管生成。出乎意料的是，VEGF应用程序也 产生由其FLT-1受体介导的明显的星形胶质细胞增殖 并增强了发育调控蛋白的神经元表达和AS 以及由其FLK-1受体介导的显着神经突生长。 S.A. 1 将检查应用于CNS外植体的VEGF的血管生成能力以及 VEGF受体的调节和血脑屏障的完整性。 S.A. 2将检验以下假设：VEGF充当星形胶质细胞和 是细胞型外植体中枢神经系统神经元的生长因子。 VEGF受体和 将使用共聚焦显微镜检查神经元和星形胶质标记。 原位杂交并通过免疫印迹和免疫亲和力量化 色谱法。创新技术利用微培养室 允许实时微透析确定VEGF是否诱导其他 细胞因子/生长因子。施用非特异性蛋白质，抗渗透 寡脱氧核苷酸和受体结合蛋白将确定 系统中的VEGF/受体特异性。我们还将启动实验 分析受体信号转导途径。 S.A. 3将测试可比 初级文化中的参数以验证VEGF对CNS的直接影响 元素，细胞类型之间没有潜在的间接作用。作为 血管生成因子VEGF可能在脑血管疾病中使用 但也可能对星形胶质细胞和神经元产生营养作用。结果可以 为理解脑组织的重要细胞生物学基础奠定重要的基础 可能会对基于VEGF的治疗做出反应。