STIM1-dependent calcium signaling in neuronal responses to hypoxia and ischemia

STIM1 依赖性钙信号传导在神经元对缺氧和缺血反应中的作用

• 批准号: 9036161
• 负责人: PAUL B ROSENBERG
• 金额: $ 23.85万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-15 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9036161
• 关键词: Acute; Address; Ataxia; Biochemical; Brain; Brain Hypoxia-Ischemia; Brain region; Calcium; Calcium Signaling; Cardiopulmonary Resuscitation; Cause of Death; Cell Death; Cell physiology; Cells; Cerebellar Diseases; Cerebellum; Cessation of life; Dependence; Event; Foundations; Functional disorder; Goals; Heart Arrest; Hippocampus (Brain); Homeostasis; Hypoxia; Image; Impaired cognition; Impairment; In Vitro; Injury; Ischemia; Knock-out; Light; Mediating; Mediator of activation protein; Modeling; Motor; Mus; Mutant Strains Mice; Neurologic; Neurologic Dysfunctions; Neuronal Injury; Neurons; Oxygen; Physiological; Property; Purkinje Cells; Role; Signal Pathway; Signal Transduction; Slice; Stroke; Synapses; Temperature; Testing; Therapeutic Intervention; Work; base; behavioral study; cell injury; deprivation; disability; human STIM1 protein; in vivo; inhibitor/antagonist; motor disorder; motor impairment; mutant; natural hypothermia; neuroprotection; novel; public health relevance; relating to nervous system; response

 DESCRIPTION (provided by applicant): Cardiac arrest and stroke are major causes of death and disability and often result in neurological impairment. Neural damage is often heterogeneous as some regions of the brain are more vulnerable to the hypoxia and ischemia (H-I) caused by these insults. The cerebellum is such a region and hypoxic and ischemic events in the cerebellum often result in ataxia and other problems with motor coordination. Neuronal calcium (Ca2+) dysregulation and Ca2+ overload are recognized as causes of cell death following brain H-I. Recently we identified stromal interaction molecules (STIM) as key components of Ca2+ signaling in excitable cells where they activate refilling of Ca2+ stores during repetitive electrical activity. Given an exceptionally high level of STIM1 expression in cerebellar Purkinje neurons and the selective vulnerability of these neurons to damage by H-I, we hypothesize that STIM1 is essential to refill Ca2+ stores and sustain Ca2+ signaling in Purkinje neurons during periods of intense synaptic activity, as occurs with H-I injury; thus, STIM1-SOCE is a critical mediator of the excitotoxic Ca2+ dysregulation and overload that causes Purkinje neuron injury and death. Our goal is to test this hypothesis by determining the fundamental properties and functions of STIM1-SOCE in Purkinje neurons during physiological and H-I conditions. For this purpose, we will carry out Ca2+ imaging, electrophysiological, morphological, and biochemical studies of cerebellar properties and function, with a focus on Purkinje neurons, in acute brain slices (Aims 1 and 2) and Purkinje neurons and other vulnerable neurons in vivo (Aim 3) in WT and STIM1 mutant mice to address the following specific aims: 1) Determine the fundamental properties and physiological functions of STIM1-dependent SOCE in cerebellar Purkinje cells, 2) Determine the contribution of STIM1-SOCE to H-I induced injury and death of Purkinje neurons in vitro, and 3) Determine the contribution of STIM1 to the neuronal damage and motor and cognitive dysfunction produced by global ischemia in vivo. These studies will begin to elucidate the role of STIM1-SOCE in Purkinje cell Ca2+ homeostasis and cerebellar function under normal physiological conditions and the contribution of STIM1-SOCE to Ca2+ overload, neuronal death, and neurological dysfunction that is produced by hypoxia and ischemia in the cerebellum and other brain regions.

 描述（由申请人提供）：心脏骤停和中风是死亡和残疾的主要原因，通常会导致神经功能障碍。神经损伤通常是异质的，因为大脑的某些区域更容易受到这些损伤引起的缺氧和缺血（H-I）的影响。小脑就是这样一个区域，小脑中的缺氧和缺血事件经常导致共济失调和其他运动协调问题。神经元钙（Ca ~（2+））失调和Ca ~（2+）超载被认为是脑缺血后细胞死亡的原因。最近，我们确定了基质相互作用分子（STIM）作为可兴奋细胞中Ca 2+信号传导的关键成分，在重复电活动期间，它们激活Ca 2+储存的再填充。鉴于小脑浦肯野神经元中STIM 1表达水平异常高，以及这些神经元对H-I损伤的选择性脆弱性，我们假设STIM 1对于在强烈突触活动期间重新填充浦肯野神经元中的Ca 2+储存和维持Ca 2+信号传导是必不可少的，如H-I损伤所发生的;因此，STIM 1-SOCE是引起浦肯野神经元损伤和死亡的兴奋毒性Ca 2+失调和过载的关键介质。我们的目标是通过确定生理和H-I条件下浦肯野神经元中STIM 1-SOCE的基本性质和功能来验证这一假设。为此，我们将在急性脑切片中进行小脑特性和功能的钙离子成像、电生理、形态学和生物化学研究，重点是浦肯野神经元（目的1和2）和浦肯野神经元和其他脆弱神经元（目的3）在WT和STIM 1突变小鼠体内的研究，以解决以下具体目标：1）确定小脑浦肯野细胞中STIM 1依赖性SOCE的基本性质和生理功能，2）确定STIM 1-SOCE对体外H-I诱导的浦肯野神经元损伤和死亡的贡献，和3）确定STIM 1在体内对由全脑缺血产生的神经元损伤以及运动和认知功能障碍的贡献。这些研究将开始阐明STIM 1-SOCE在正常生理条件下浦肯野细胞Ca 2+稳态和小脑功能中的作用，以及STIM 1-SOCE对小脑和其他脑区缺氧和缺血引起的Ca 2+过载、神经元死亡和神经功能障碍的贡献。