Brain Injury and Intercelluar Calcium Waves

描述（由申请人提供）：当前提案的目标是 评估创伤性脑损伤改变细胞间 星形胶质细胞之间的通信，一个潜在的关键，往往被忽视 大脑处理的一部分。星形胶质细胞曾被视为被动细胞， 现在被认为是大脑中信号处理的积极贡献者。 然而，创伤性损伤对钙介导的星形胶质细胞细胞间 没有评估信令。因此，建议对 创伤性损伤改变细胞间钙信号传导的假说 星形胶质细胞的神经胶质合胞体中使用以下具体目标： 具体目标1：皮质星形胶质细胞损伤改变起源和分化， 细胞间钙波的传播。我们将比较速度和 损伤星形胶质细胞胞间钙波与 未受伤的星形胶质细胞（方法：采用定量荧光显微镜， 评价细胞间钙波）。 具体目标2：损伤改变间隙连接偶联。我们将比较 荧光染料通过间隙连接转移，连接蛋白43的表达 蛋白，以及未损伤和 受损的星形胶质细胞方法：用荧光黄转移法测定偶联度， 缝隙连接;使用蛋白质印迹来测量 连接蛋白43蛋白）。 具体目标3：细胞间钙信号传导将维持在 通过药理学操作IP 3介导的细胞内 钙信号（方法：使用元件的激动剂和拮抗剂， 信号通路，以确定细胞间钙波的改变 可以被削弱）。 这些拟议的实验首次评估了 创伤性脑损伤对复杂而广泛的钙信号网络的影响 在星形胶质细胞中。目的#1直接检查机械损伤对 星形胶质细胞中的细胞间钙信号。目标#2检查潜力 机制（即缝隙连接）参与损伤诱导的改变， 细胞间钙信号目标#3检查潜在治疗 可以恢复细胞间信号传导的干预措施。的结果 拟议的实验不仅可以提供新的见解， TBI，但也可能导致新的治疗方法的发展， 治疗创伤性脑损伤的方法。

DESCRIPTION (provided by applicant): The goal of the current proposal is to evaluate the hypothesis that traumatic brain injury alters intercellular communication between astrocytes, a potentially crucial and often overlooked aspect of brain processing. Astrocytes were once viewed as passive cells, but are now characterized as active contributors to signal processing in the brain. Yet, the effect of traumatic injury on calcium-mediated astrocyte intercellular signaling has not been evaluated. Therefore, the proposal evaluates the hypothesis that traumatic injury alters intercellular calcium signaling between astrocytes in the glial syncytium using the following specific aims: Specific Aim 1: Injury of cortical astrocytes alters origination and propagation of intercellular calcium waves. We will compare the velocity and distance of intercellular calcium waves in injured astrocytes to that of uninjured astrocytes. (Method: Use quantitative fluorescent microscopy to evaluate intercellular calcium waves). Specific Aim 2: Injury alters gap junction coupling. We will compare fluorescent dye transfer through gap junctions, expression of Connexin 43 protein, and phosphorylation of Connexin 43 protein between uninjured and injured astrocytes. (Method: Use lucifer yellow transfer to measure coupling of gap junctions; use Western Blots to measure expression and phosphorylation of Connexin 43 protein). Specific Aim 3: Intercellular calcium signaling will be maintained in astrocytes by pharmacological manipulation of IP3-mediated intracellular calcium signaling. (Method: Use agonists and antagonists of elements in the signaling pathway to determine if alterations in intercellular calcium waves can be attenuated). These proposed experiments evaluate, for the first time, the effect of traumatic brain injury on the intricate and extensive calcium signaling network among astrocytes. Aim #1 directly examines the effects of mechanical injury on intercellular calcium signaling in astrocytes. Aim #2 examines the potential mechanisms (i.e. gap junctions) involved in injury-induced alterations in intercellular calcium signaling. Aim #3 examines potential therapeutic interventions which could restore intercellular signaling. The results of the proposed experiments may provide not only new insights into the pathophysiology of TBI, but also potentially lead to the development of novel therapeutic approaches for the treatment of the traumatically injured brain.