Acid-sensing ion channels and ischemic brain injury

酸敏感离子通道与缺血性脑损伤

• 批准号: 7426789
• 负责人: ROGER Pancoast SIMON
• 金额: $ 33.33万
• 依托单位: LEGACY EMANUEL HOSPITAL AND HEALTH CENTER
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-24 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7426789
• 关键词: ASIC channel; Acidosis; Acids; Affect; Affinity; Amiloride; Apoptotic; Attention; Attenuated; Brain; Brain Injuries; Brain Ischemia; Cell Culture System; Cell Death; Cell Death Process; Cell Line; Cells; Chemosensitization; DNA Damage; Data; Diffuse; Effectiveness; Equilibrium; Excitatory Amino Acid Antagonists; Family; Gated Ion Channel; Glucose; Glutamates; Human; Image; Individual; Infarction; Injury; Ion Channel; Ischemia; Ischemic Brain Injury; Ischemic Stroke; Knockout Mice; Learning; Lysine; Mediating; Mediator of activation protein; Membrane; Memory; Metabolic; Methods; Modeling; Molecular; Mus; Mutation; Necrosis; Nervous System Physiology; Neuraxis; Neuronal Injury; Neurons; Neurotoxins; Oxygen; Patch-Clamp Techniques; Pathologic; Permeability; Play; Population; Process; Proteins; Publishing; Range; Rattus; Research Personnel; Role; Site; Site-Directed Mutagenesis; Stimulation of Cell Proliferation; Stimulus; Stress; Stroke; Synapses; Synaptic plasticity; System; Techniques; Testing; Time; Toxic effect; Transfection; base; cell injury; deprivation; extracellular; falls; in vitro Model; in vivo; member; neuroprotection; neurotransmission; novel; perpetrators; protective effect; receptor; research study; response; stroke therapy; uptake

DESCRIPTION (provided by applicant): Brain ischemia is characterized by a marked, rapid fall in pH, which is assumed to be injurious although through multiple or uncertain mechanisms. The recent discovery in brain of Acid Sensing Ion Channels (ASICs), which are ubiquitous and function physiologically in synaptic neurotransmission, offers a diffuse, membrane based, receptor gated ion channel system, which will respond to the pathologic pH fall in brain ischemia. These H+ receptor gated channels are Na+ channels, a portion of which is also Ca2+ permeable. Acid sensitivity and Ca2+ permeability suggest a role in ischemic brain injury. Using patch clamp techniques and Ca2+ imaging of native neurons in cortical cultures we show the pH sensitivity of these channels and acid induced Ca2+ uptake. Both the acid induced channel currents and Ca2+ uptake are greatly potentiated in the setting of modeled ischemia (Oxygen Glucose Deprivation-OGD or NaCN). Thus these channels respond to both acidosis and "ischemia" in a mutually potentiating manner. The channel current and Ca2+ uptake are glutamate independent, are inhibited by ASIC pharmacologic blockade and by specific blockade of the ASIC 1a channel subunit. Thus, our preliminary studies support new cellular and molecular mechanisms mediating ischemic-acidosis induced brain injury, which we offer to dissect with the following Specific Aims: Single cell recording and transfection of individual ASIC subunit cDNAs will show that: 1) Ca2+ permeable ASICs produce cell injury in modeled ischemia via a subunit specific mechanism. Using rat and mouse global ischemia models and ASIC1a & ASIC2a knockout mice, we propose that: 2) Blockade of ASICs protects against ischemic brain injury in vivo. With molecular techniques including site directed mutagenesis we will describe endogenous Zn2+ modulation of Ca2+permeable ASICs: 3) Characterize high-affinity Zn2+ modulation of ASIC currents in acidosis and ischemia -induced cell injury. These experiments will describe new, glutamate independent, mechanisms of ischemic brain injury and the central role of ischemic-acidosis. ASIC blockade will offer new and potent potential therapy for stroke.

描述（由申请人提供）：脑缺血的特征是pH值显著、快速下降，尽管通过多种或不确定的机制，但被认为是有害的。近年来在脑中发现的酸敏感离子通道（Acid Sensing Ion Channels，ASIC）是一种弥散的、基于膜的、受体门控的离子通道系统，它能对脑缺血时病理性的pH下降作出反应。这些H+受体门控通道是Na+通道，其中一部分也是Ca 2+渗透性的。酸敏感性和Ca 2+渗透性表明在缺血性脑损伤中的作用。使用膜片钳技术和Ca 2+成像的本地神经元在皮层文化中，我们显示这些通道的pH敏感性和酸诱导的Ca 2+摄取。酸诱导的通道电流和Ca 2+摄取在模型缺血（氧葡萄糖剥夺-OGD或NaCN）的设置中大大增强。因此，这些通道以相互增强的方式对酸中毒和“缺血”作出反应。通道电流和Ca 2+摄取是谷氨酸非依赖性的，被ASIC药理学阻断和ASIC 1a通道亚基的特异性阻断抑制。因此，我们的初步研究支持新的细胞和分子机制介导的缺血酸中毒诱导的脑损伤，我们提供了解剖与以下具体目的：单个细胞记录和转染的ASIC亚基cDNA将显示：1）Ca 2+渗透ASIC通过亚基特异性机制在模型缺血中产生细胞损伤。利用大鼠和小鼠全脑缺血模型以及ASIC 1a和ASIC 2a基因敲除小鼠，我们提出：2）阻断ASICs对体内缺血性脑损伤具有保护作用。利用包括定点诱变的分子技术，我们将描述Ca 2+可渗透ASIC的内源性Zn 2+调节：3）表征酸中毒和缺血诱导的细胞损伤中ASIC电流的高亲和力Zn 2+调节。这些实验将描述新的，谷氨酸独立，缺血性脑损伤的机制和缺血性酸中毒的核心作用。ASIC阻断将为脑卒中提供新的、潜在的治疗方法。