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

基本信息

批准号：
8722056
负责人：
ZONGLU S HUA
金额：
$ 19.69万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-01 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8722056
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

项目摘要

DESCRIPTION (provided by applicant): 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.

描述（由申请人提供）：创伤性脑损伤（TBI）是由外部机械力（例如爆炸或碰撞）造成的脑损伤。我们目前对 TBI 的理解主要来自体内研究，这些研究显示了 TBI 后取样细胞可测量的生物学效应。人们对损伤性刺激期间脑细胞的主要机械应力以及它们如何转化为 TBI 涉及的细胞和分子事件知之甚少。该提案旨在研究哪些刺激特性对细胞损伤最关键，以及机械刺激如何与次级细胞过程耦合。我们将在由快速压力伺服驱动的微流体室中使用组织培养的成年星形胶质细胞来产生时间依赖性流体剪切，可以模拟爆炸产生的剪切应力。我们将使用基因编码的荧光应激传感器直接测量特定细胞骨架蛋白的应激。该研究有两个具体目标。目标 1 确定导致长期变化的刺激的物理特性。通过使用不同振幅、上升时间和频率的冲击波测量特定细胞骨架蛋白中应力的时间过程，我们将确定导致不可逆变形的参数。这种不可逆性将通过研究解剖结构和应力分布的变化来确定。目标 2 研究细胞在机械损伤期间和之后的反应，以确定机械刺激如何改变细胞稳态。为此，我们将测量细胞内 Ca2+ 和细胞体积的时间依赖性变化。这些特性将与细胞应激同时测量以确定因果关系。我们将使用特异性抑制剂 GsMTx4 进一步测试机械敏感通道在传递 Ca2+ 方面的作用，并探讨预先给予 GsMTx4 是否可以用作预防性治疗。