Vascular Pannexin 1 in Ischemic Stroke

|| 项目总结/摘要 缺血性卒中是一种可导致神经功能缺损或死亡的重要疾病;然而， 药理学干预药物是可用的，其主要目的是消除物理闭塞 在血管内。探索潜在的治疗靶点，以改善缺血性脑卒中后的恢复， 中风时，我们开始研究Panx 1通道的作用，它激活下游嘌呤能受体， 在大脑血管系统中。我最近的研究表明，内皮Panx 1基因的缺失， 脑缺血/再灌注后梗死体积。然而，内皮Panx 1可能显著促进 动脉和静脉内皮功能。在这个应用中，我们证明了动脉肌源性张力 发展（动脉）和缺血性卒中后白细胞炎症（静脉）， 在缺乏内皮Panx 1的小鼠中减少。该应用测试了内皮Panx 1 通过调节脑动脉肌源性神经元和神经元的表达， 紧张和调节白细胞粘附和迁移的脑静脉内皮细胞。我们的初步 数据表明，在肌源性张力发育过程中，Panx 1的激活是通过与b3相互作用来调节的， 整合素然而，在脑静脉中，我发现白细胞粘附在缺血后的脑静脉中， 中风是通过Panx 1通道功能调节的。这些研究将提供洞察力的差异作用 以及动脉和静脉脑内皮细胞内Panx 1的调节，它们对脑血管病病因的贡献 缺血性卒中损伤，并为未来确定缺血性卒中的治疗干预措施奠定基础 中风患者 在本申请的指导阶段，我将开发新的转基因小鼠，以评估 过度表达内皮Panx 1，并获得必要的技能，以评估功能的后果， Panx 1-b3相互作用和使用体内活体内成像的缺血性卒中后的脑炎症 显微镜技术。我将继续我的专业和科学发展， 我的指导委员会持续提供指导。在独立阶段，我 将开发一种新的转基因小鼠，将删除或过表达Panx 1特异性动脉内 内皮细胞和确定动脉与静脉内皮Panx 1在缺血性中风中的作用。 此外，我将建立下游嘌呤能信号参与Panx 1依赖性白细胞 粘附，通过脑小静脉的体内成像，和迁移，使用流式细胞术，及其对 确定缺血性中风的严重程度。这个项目将促进我继续科学和专业 培训和促进我在学术界建立一个独立的研究计划。

|| 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.