Traumatic brain injury: early mechanosensitive events in astrocytes

创伤性脑损伤：星形胶质细胞的早期机械敏感事件

• 批准号: 8622510
• 负责人: ZONGLU S HUA
• 金额: $ 23.85万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8622510
• 关键词: Accidents; Actinin; Address; Adult; Anatomy; Astrocytes; Biological; Biological Assay; Blast Cell; Brain; Brain Injuries; Cardiovascular system; Cell Volumes; Cell physiology; Cells; Cellular Stress; Cellular Structures; Complex; Coupled; Coupling; Cytoskeletal Proteins; Cytoskeleton; Dependence; Disease; Etiology; Event; Fluorescence Resonance Energy Transfer; Fluorescent Probes; Focal Adhesions; Frequencies; Glial Fibrillary Acidic Protein; Global Change; Homeostasis; Hour; Injury; Lead; Link; Liquid substance; Maintenance; Measurable; Measures; Mechanical Stress; Mechanics; Medicine; Metabolic; Microfluidics; Molecular; Neurons; Neurotransmitters; Pharmaceutical Preparations; Pharmacotherapy; Physiologic pulse; Play; Preventive; Process; Property; Proteins; Research; Role; Sampling; Secondary to; Solutions; Spectrin; Stimulus; Stress; Structure; Subcellular Anatomy; Synapses; Talin; Testing; Therapeutic; Therapeutic Agents; Time; Traumatic Brain Injury; Variant; Veterans; Vinculin; brain cell; cell injury; in vivo; inhibitor/antagonist; paxillin; physical property; pressure; prevent; public health relevance; research study; response; sensor; shear stress; tissue culture

Abstract Traumatic brain injury (TBI) is brain damage resulting from an external mechanical force, such as blast or crashes. Our current understanding of TBI is derived mainly from in vivo studies that show measurable biological effects on cells sampled after TBI. Little is known about the primary mechanical stresses in brain cells during damaging stimuli, and how they are transduced into cellular and molecular events involved in TBI. This proposal aims to examine what stimulus properties are most critical to cell injury and how the mechanical stimulus is coupled to secondary cellular processes. We will use tissue cultured adult astrocytes in a microfluidic chamber driven by a fast pressure servo to generate time dependent fluid shear that can be made to emulate shear stress from a blast. We will directly measure the stresses in specific cytoskeletal proteins using genetically encoded fluorescent stress sensors. The research has two specific aims. Aim 1 identifies physical properties of the stimulus that lead to long term alterations. By measuring the time course of stresses in specific cytoskeletal proteins using shockwaves of various amplitude, rise time, and frequency, we will determine the parameters that lead to irreversible deformation. This irreversibility will be determined by investigating changes in anatomy and stress distribution. Aim 2 investigates the cells' response during and immediately after mechanical insult to determine how mechanical stimulus alters cell homeostasis. For this we will measure the time dependent changes of intracellular Ca2+ and cell volume. These properties will be measured simultaneously with cellular stress to determine causality. We will further test the role of mechanosensitive channels for passing Ca2+ using specific inhibitor, GsMTx4, and explore whether pre-administration of GsMTx4 might be used as a preventive therapy.

抽象的 创伤性脑损伤（TBI）是由外部机械力造成的脑损伤，例如 如爆炸或碰撞。我们目前对 TBI 的理解主要来自体内研究： 对 TBI 后取样的细胞显示出可测量的生物学效应。对初级知之甚少 破坏性刺激期间脑细胞的机械应力，以及它们如何转化为 TBI 涉及的细胞和分子事件。该提案旨在研究什么刺激措施 特性对于细胞损伤以及机械刺激如何与细胞损伤相结合至关重要 次级细胞过程。我们将在微流体中使用组织培养的成年星形胶质细胞 腔室由快速压力伺服系统驱动，产生随时间变化的流体剪切力 用于模拟爆炸产生的剪切应力。我们将直接测量特定的应力 使用基因编码的荧光应激传感器的细胞骨架蛋白。该研究有 两个具体目标。目标 1 确定导致长期结果的刺激的物理特性 变更。通过使用以下方法测量特定细胞骨架蛋白中应力的时间过程 各种振幅、上升时间和频率的冲击波，我们将确定参数 从而导致不可逆的变形。这种不可逆转性将通过调查来确定 解剖结构和应力分布的变化。目标 2 研究细胞在期间和期间的反应 机械损伤后立即确定机械刺激如何改变细胞 体内平衡。为此，我们将测量细胞内 Ca2+ 和细胞内 Ca2+ 的时间依赖性变化。 体积。这些特性将与细胞应激同时测量，以确定 因果关系。我们将使用以下方法进一步测试机械敏感通道在传递 Ca2+ 方面的作用： 特异性抑制剂 GsMTx4，并探索 GsMTx4 的预给药是否可以用作 一种预防性疗法。