Invivo Spatio Temporal Optical Imaging of Focal Ishemia

描述(申请人摘要)：这项建议，“活体时空 局灶性脑缺血的光学成像》，是对NINDS的回应 辅导型定量研究职业发展计划公告 奖项(K25)。应聘者希望获得FOCUS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 半影区可能是可以挽救的，重要的是要量化血液 体内半暗带内的流动变化和分子事件。在本建议书中 将使用高分辨率光学成像技术来评估 局灶性脑缺血后体内半影区的病理生理变化。 具体地说，导致血液动力学和细胞事件的改变 病灶后将检查微血管形成或凋亡细胞死亡。 缺血性脑损伤。血液的时空特征 大鼠从核心到半影区再到正常组织转变过程中的血流变化 局灶性缺血模型将使用二维激光在活体内进行量化 斑点血流成像。血流量的时空变化 在缺血半暗带中，eNOS会动态上调 量过了。在描述了半影区的血流变化后， 作为细胞凋亡指标的caspase-3活性水平将是 高表达重组GFP半胱氨酸天冬氨酸氨基转移酶小鼠的体内检测 用双光子荧光显微镜观察底物。这种独特的动物模型 将允许体内可视化和量化半胱氨酸天冬氨酸酶的作用 抑制剂(DEVD-fmk)对全脑缺血时caspase-3裂解水平的影响 半影区。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 将先进的光学成像技术应用于 研究缺血半暗带。

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.