Brain Injury and Intercelluar Calcium Waves

脑损伤和细胞间钙波

• 批准号: 6622425
• 负责人: CANDACE L. FLOYD
• 金额: $ 4.64万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-06-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6622425
• 关键词: Animalia; astrocytes; brain injury; calcium flux; calcium indicator; cell cell interaction; fluorescence microscopy; gap junctions; gene expression; membrane channels; neural transmission; neurogenetics; neuropharmacologic agent; neuropharmacology; neuroregulation; phosphorylation; postdoctoral investigator; protein structure function; time resolved data; western blottings

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.

描述（申请人提供）：当前建议的目的是 评估创伤性脑损伤改变细胞间的假设 星形胶质细胞之间的沟通，这是一种潜在的至关重要的，经常被忽略的 大脑处理的方面。星形胶质细胞曾经被视为被动细胞，但 现在被认为是大脑信号处理的主动贡献者。 然而，创伤性损伤对钙介导的星形胶质细胞间细胞的影响 信号尚未评估。因此，提案评估了 创伤性损伤改变细胞间钙信号的假设 使用以下特定目的，神经胶质合成中的星形胶质细胞： 特定目的1：皮质星形胶质细胞的损伤改变了起源和 细胞间钙波的传播。我们将比较速度和 受伤的星形胶质细胞中细胞间钙波的距离与 未受伤的星形胶质细胞。 （方法：使用定量荧光显微镜 评估细胞间钙波）。 特定目标2：伤害改变间隙连接耦合。我们将比较 通过间隙连接的荧光染料转移，连接蛋白的表达43 蛋白质和连接蛋白43蛋白在未受伤和 受伤的星形胶质细胞。 （方法：使用路西法黄色转移来测量 间隙连接；使用蛋白质印迹测量表达和磷酸化 连接蛋白43蛋白）。 特定的目标3：细胞间钙信号传导将保持在 通过IP3介导的细胞内的药理学操纵，星形胶质细胞 钙信号传导。 （方法：使用激动剂和拮抗剂 确定细胞间钙波是否改变的信号通路 可以减弱）。 这些提出的实验首次评估了 复杂而广泛的钙信号网络上的创伤性脑损伤 在星形胶质细胞中。 AIM＃1直接检查机械损伤对 星形胶质细胞中的细胞间钙信号传导。 AIM＃2检查潜力 涉及伤害引起的改变的机制（即间隙连接） 细胞间钙信号传导。 AIM＃3检查潜​​在的治疗性 可以恢复细胞间信号的干预措施。结果 拟议的实验不仅可以为病理生理学提供新的见解 TBI，但也有可能导致新型治疗的发展 治疗创伤受伤的大脑的方法。