Invivo Spatio Temporal Optical Imaging of Focal Ishemia

局灶性缺血的 Invivo 时空光学成像

• 批准号: 6612810
• 负责人: Andrew K Dunn
• 金额: $ 14.62万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-15 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6612810
• 关键词: apoptosis; autoradiography; bioimaging /biomedical imaging; blood flow measurement; brain imaging /visualization /scanning; brain mapping; cerebral ischemia /hypoxia; cysteine endopeptidases; disease /disorder model; enzyme activity; enzyme inhibitors; enzyme substrate; fluorescence microscopy; fluorescence resonance energy transfer; green fluorescent proteins; hemodynamics; hypercapnia; laboratory mouse; laboratory rat; molecular pathology; neurophysiology; nitric oxide synthase; technology /technique development; temporal lobe /cortex; tissue /cell culture

DESCRIPTION (Applicant's Abstract): This proposal, "In vivo Spatio-Temporal Optical Imaging of Focal Ischemia", is submitted in response to the NINDS program announcement for Mentored Quantitative Research Career Development Awards (K25). The candidate seeks to gain training in the neurobiology of focal cerebral ischemia in order to apply advanced optical imaging techniques to the study of cerebral ischemia in animal models. Since tissue in the ischemic penumbra can potentially be salvaged, it is important to quantify the blood flow changes and molecular events within the penumbra in vivo. In this proposal high resolution optical imaging techniques will be used to assess pathophysiologic events in the penumbra in vivo following focal ischemia. Specifically, the hemodynamic and cellular events that lead to alterations in the microvasculature or apoptotic cell death will be examined following focal ischemic brain injury. The spatial and temporal characteristics of the blood flow changes in the transition from core to penumbra to normal tissue in a rat model of focal ischemia will be quantified in vivo using two-dimensional laser speckle blood flow imaging. The spatial and temporal alterations in blood flow in the ischemic penumbra in response to eNOS upregulation will be dynamically measured. Having characterized the blood flow changes in the penumbra, the level of caspase-3 activity, which is an indicator of apoptosis, will be measured in vivo in mice overexpressing recombinant GFP-based caspase substrates with two-photon fluorescence microscopy. This unique animal model will allow in vivo visualization and quantification of the effects of a caspase inhibitor (DEVD-fmk) on the level of caspase-3 cleavage throughout the ischemic penumbra. Through the training that this award provides, the candidate will develop the knowledge of neuroscience and stroke physiology necessary to lead an independent research career applying advanced optical imaging techniques to study the ischemic penumbra.

描述（申请人的摘要）：该提议，“体内空间-时间 局灶性缺血的光学成像”，是对NINDS的回应 导师定量研究职业发展计划公告 奖项（K25）。候选人寻求获得局灶性神经生物学方面的培训， 为了将先进的光学成像技术应用于脑缺血， 脑缺血动物模型的研究因为缺血区的组织 半暗带可能被挽救，重要的是要量化血液 体内半影区内的流动变化和分子事件。本提案中 高分辨率光学成像技术将用于评估 局灶性缺血后体内半影区的病理生理事件。 具体来说，血液动力学和细胞事件，导致改变， 微血管或凋亡性细胞死亡将在病灶后进行检查， 缺血性脑损伤血液的时空特征 大鼠从核心区到半影区再到正常组织过渡过程中的血流变化 将使用二维激光在体内定量局灶性缺血模型 斑点血流成像。血流的空间和时间变化 在缺血半暗带中，响应eNOS上调将动态地 测定了在描述了半暗带中的血流变化之后， 作为细胞凋亡指标的半胱天冬酶-3活性水平， 在过表达基于GFP的重组胱天蛋白酶的小鼠体内测量 用双光子荧光显微镜观察基质。这个独特的动物模型 将允许半胱天冬酶的作用的体内可视化和定量 抑制剂（DEVD-fastin）对整个缺血性脑损伤中caspase-3裂解水平的影响 半影。通过该奖项提供的培训，候选人将 发展必要的神经科学和中风生理学知识， 一个独立的研究生涯应用先进的光学成像技术， 研究缺血半暗带