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

基因转移方法是一种新的，潜在的有价值的方法 研究血管生物学基因转移到脑血管， 然而，这是极其困难。 调查人员描述说， 第一次基因转移到颅内血管， 血管周围组织，这是通过注射完成的 复制缺陷型腺病毒载体进入脑脊髓 液 研究人员使用腺病毒表达β- 半乳糖苷酶，报告基因的产物。 一个重要的下一步 基因改变颅内血管功能 转移 该项目的目标是改变血管功能， 通过基因转移， 正常动物内皮型一氧化氮合酶（eNOS）， eNOS敲除小鼠。研究人员已经克隆了 将eNOS导入腺病毒载体中。 在试图改变 在体内脑血管的血管功能， 研究人员将尝试增强eNOS介导的 通过基因转移到体外颈动脉的血管舒张。 初步研究表明，这种方法是可行的。 的 关键的下一步是发展基因转移方法， 脑循环改变脑血管功能 体内的血管 建议进行研究，以确定是否 在体内将eNOS基因转移到脑血管产生 颈动脉中NO介导的刺激增强的血管舒张 动脉、颅内大脑小动脉和基底动脉。 通过基因检测NO介导的应答的改善 在eNOS敲除小鼠中可促进eNOS的转移。 计划在eNOS敲除小鼠中进行研究，以确定是否 将eNOS基因转移到主动脉和颈动脉， 介导的反应和是否基因转移eNOS， eNOS敲除小鼠体内颅内血管减少 补偿机制的作用。 施行本 研究是双重的：开发基因转移的方法， 脑血管，并使用这些方法作为研究的工具， 血管生物学

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.