Vascular Pannexin 1 in Ischemic Stroke

|项目摘要/摘要 缺血性中风是一种重要的疾病，可导致神经功能障碍或死亡；然而，只有一种 药物干预药物是可用的，其主要目的是消除物理闭塞。 在血管内。探讨提高脑缺血后恢复的潜在治疗靶点 在中风时，我们开始研究Panx1通道的作用，Panx1通道激活下游的嘌呤能受体， 在脑血管系统内。我最近证明，删除内皮细胞Panx1可以显著改善 脑缺血/再灌流后脑梗塞体积。然而，内皮细胞Panx1可以显著促进 动脉和静脉内皮细胞都有功能。在本应用中，我们演示了动脉肌源性张力 缺血性卒中后白细胞的发育(动脉)和炎症(静脉)显著 在缺乏内皮细胞Panx1的小鼠中降低。这个应用程序测试了内皮细胞Panx1 脑动脉肌源性调节参与缺血性卒中损伤的发生发展 脑静脉内皮细胞白细胞黏附和迁移的张力及调节。我们的预赛 数据表明，在肌张力发育过程中Panx1的激活是通过与b3的相互作用来调节的。 整合素。然而，在脑静脉中，我发现脑缺血后脑微静脉中的白细胞粘连 行程通过Panx1通道功能进行调节。这些研究将提供对不同角色的洞察 Panx1在动、静脉脑内皮细胞中的表达及其在脑缺血发病机制中的作用 缺血性卒中损伤，并为未来确定缺血性治疗干预措施奠定了基础 中风患者。 在这项应用的指导阶段，我将开发新的转基因小鼠来评估 内皮细胞Panx1的过表达并获得必要的技能以评估其功能后果 活体内研究Panx1-b3相互作用与缺血性卒中后显像性脑炎症 显微镜技术。我将继续我的专业和科学发展，为 在我的指导委员会的持续指导下，进入了独立阶段。在独立阶段，我 将开发一种新的转基因小鼠，将在动脉中特异地缺失或过度表达Panx1 并确定动脉和静脉内皮细胞Panx1在缺血性中风中的作用。 此外，我还将建立参与Panx1依赖的白细胞的下游嘌呤能信号。 黏附，通过活体成像的脑小静脉，和移行，使用流式细胞术，及其影响 确定缺血性中风的严重程度。这个项目将培养我继续科学和专业的能力 培训和促进我在学术界建立一个独立的研究项目。

|| PROJECT SUMMARY/ABSTRACT Ischemic stroke is a significant disease that can lead to neurological deficits or death; however, only one pharmacological intervention drug is available and it predominately aims to eliminate the physical occlusion within the blood vessel. To explore potential therapeutic targets to improve recovery following an ischemic stroke, we began examining the role of Panx1 channels, which activate downstream purinergic receptors, within the cerebral vasculature. I have recently shown that deletion of endothelial Panx1 profoundly improves post-cerebral ischemia/reperfusion infarct volume. However, endothelial Panx1 could significant contribute to both arterial and venous endothelial functions. In this application, we demonstrate that arterial myogenic tone development (arterial), and inflammation of leukocytes following an ischemic stroke (venous), are significantly decreased in mice lacking endothelial Panx1. This application tests the hypothesis that endothelial Panx1 contributes to the development of ischemic stroke injury through both regulation of cerebral arterial myogenic tone and regulation of leukocyte adhesion and emigration in cerebral venous endothelial cells. Our preliminary data suggest that Panx1 activation during myogenic tone development is regulated through interaction with b3 integrins. Whereas, in cerebral veins, I find leukocyte adhesion in cerebral venules following an ischemic stroke is regulated through Panx1 channel function. These studies will provide insight into the differential roles and regulation of Panx1 within arterial and venous cerebral endothelial cells, their contribution to the etiology of ischemic stroke injury, and lay the foundation for future identification of therapeutic interventions for ischemic stroke patients. During the mentored phase of this application, I will develop novel transgenic mice to evaluate the overexpression of endothelial Panx1 and obtain necessary skills to evaluate the functional consequence of Panx1-b3 interaction and image cerebral inflammation following an ischemic stroke using in vivo intravital microscopy techniques. I will continue my professional and scientific development in preparation for the independent phase with continuous guidance from my mentoring committee. During the independent phase, I will develop a novel transgenic mouse that will delete or overexpress Panx1 specifically within arterial endothelial cells and determine the role of arterial verse venous endothelial Panx1 during an ischemic stroke. In addition, I will establish the downstream purinergic signaling involved in Panx1-dependent leukocyte adhesion, by in vivo imaging of cerebral venules, and emigration, using flow cytometry, and its impact of determining the severity of an ischemic stroke. This project will foster my continued scientific and professional training and facilitate my establishing an independent research program in academia.