PHYSIOLOGICAL REGUALTION OF CEREBRAL CIRCULATION--GENE TRANSFER OF NITRIC OXIDE S

脑循环的生理调节--一氧化氮S的基因转移

• 批准号: 6243609
• 负责人: DONALD D HEISTAD
• 金额: $ 16.17万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-06-10 至 1998-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6243609
• 关键词: Adenoviridae; cardiovascular pharmacology; carotid artery; cell transformation; forskolin; gene expression; genetic promoter element; genetically modified animals; histochemistry /cytochemistry; laboratory mouse; molecular cloning; nitric oxide synthase; tissue /cell culture; transfection; transfection /expression vector; vascular endothelium; vasomotion

Gene transfer approaches are a novel, potentially valuable method to study vascular biology. Gene transfer to cerebral blood vessels, however, is extremely difficult. The investigators have described the first gene transfer to intracranial blood vessels and perivascular tissue in vivo, which was accomplished by injection of a replication-deficient adenovirla vector into cerebrospinal fluid. The investigators have used adenovirus that expresses beta- galactosidase, the product of a reporter gene. A major next step will be alteration of function of intracranial blood vessels by gene transfer. The goal of this project is to alter vasomotor function of the carotid artery and intracranial vessels by gene transfer of endothelial nitric oxide synthase (eNOS) in normal animals and eNOS knockout mice. The investigators have cloned the gene for eNOS into an adenoviral vector. Before attempting to alter vasomotor function in cerebral blood vessels in vivo, the investigators will attempt to augment eNOS-mediated vasodilatation by gene transfer to the carotid artery in vitro. Preliminary studies suggest that this approach is feasible. The critical next step in development of gene transfer approaches to the cerebral circulation is to alter vascular function of cerebral blood vessels in vivo. Studies are proposed to determine whether gene transfer of eNOS to cerebral vessels in vivo produces augmented vasodilatation of NO-mediated stimuli in the carotid artery, intracranial cerebral arterioles, and the basilar artery. Detection of improvement of NO-mediated responses by gene transfer of eNOS may be facilitated in eNOS knockout mice. Studies are planned in eNOS knockout mice to determine whether gene transfer of eNOS to the aorta and carotid artery restores NO- mediated responses and whether gene transfer of eNOS to intracranial blood vessels in vivo in eNOS knockout mice reduces the role of compensatory mechanisms. The purpose of these studies is two-fold: to develop methods for gene transfer to cerebral vessels, and to use these approaches as a tool to study vascular biology.

基因转移方法是一种新的、有潜在价值的方法 研究血管生物学。基因转移到脑血管， 然而，这是极其困难的。调查人员描述了 第一次将基因转移到颅内血管和 体内血管周围组织，这是通过注射完成的 将复制缺陷型腺病毒载体导入脑脊液 流体。研究人员使用了表达β-蛋白的腺病毒。 半乳糖苷酶，一种报告基因的产物。下一步的重要步骤 将是由基因引起的颅内血管功能的改变 调职。这个项目的目标是改变血管舒缩功能 基因转移对颈动脉和颅内血管的影响 内皮型一氧化氮合酶(ENOS)在正常动物和 ENOS基因敲除小鼠。研究人员已经克隆了该基因用于 ENOS转化腺病毒载体。在尝试更改 活体脑血管的血管运动功能 调查人员将试图加强eNOS介导的 体外颈动脉基因转移的血管扩张作用。 初步研究表明，这种方法是可行的。这个 开发基因转移方法的关键下一步 脑循环是为了改变大脑的血管功能 体内的血管。建议进行研究以确定 内皮型一氧化氮合酶基因转移到体内脑血管产生 NO介导的刺激增强颈动脉的血管扩张作用 动脉、脑小动脉和基底动脉。 检测基因对NO介导的应答的改善 在eNOS基因敲除的小鼠中，eNOS的转移可能是容易的。 计划在eNOS基因敲除小鼠中进行研究，以确定 内皮型一氧化氮合酶基因转导大鼠主动脉和颈动脉恢复NO- 内皮型一氧化氮合酶的介导性反应及基因转移是否对 ENOS基因敲除小鼠体内颅内血管减少 补偿机制的作用。这些活动的目的是 研究有两个方面：开发基因转移的方法 脑血管，并将这些方法作为研究的工具 血管生物学。