AMYLOID IN EXCITATORY AMINO ACID-INDUCED CALCIUM NEUROTOXICITY

兴奋性氨基酸诱导的钙神经毒性中的淀粉样蛋白

• 批准号: 6234410
• 负责人: MARK P MATTSON
• 金额: $ 1.14万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-01 至 1998-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6234410
• 关键词: Alzheimer's disease; NMDA receptors; amyloid proteins; brain mapping; calcium binding protein; calcium channel; calcium flux; calcium transporting ATPase; confocal scanning microscopy; cytotoxicity; electron microscopy; excitatory aminoacid; fluorescent dye /probe; histopathology; hormone regulation /control mechanism; human tissue; immunocytochemistry; laboratory rat; membrane transport proteins; neural degeneration; neurofibrillary tangles; neurotransmitter receptor; tissue /cell culture; voltage /patch clamp

The long-term goal of this research is to gain an understanding of the cellular and molecular events that lead to neuronal damage and death in Alzheimer's disease (AD). This project tests the hypothesis that beta- amyloid destabilizes cellular calcium homeostasis and thereby renders neurons more vulnerable to environmental insults. A hippocampal cell culture system will be used to examine the cellular and molecular mechanisms whereby beta-amyloid destabilizes neuronal calcium homeostasis, and potentiates excitatory amino acid neurotoxicity. Immunolocalization studies of AD brains are designed to determine whether the neuropathology of AD is consistent with the calcium destabilization hypothesis of beta- amyloid neurotoxicity. The first aim will test the hypothesis that beta- amyloid affects specific cellular systems for calcium homeostasis (NMDA receptors, calcium channels Na+/Ca2+ exchanger, calcium binding protein and calcium ATPase). The second aim is to determine whether endogenous beta- amyloid contributes to selective neuronal vulnerability in cell culture. The third aim is to establish whether the cytoskeletal manifestations of neuronal degeneration induced by beta-amyloid resemble the neurofibrillary pathology of AD. The fourth aim is to determine the relationships of beta- amyloid, NMDA receptors, and calcium-regulating proteins in the histopathology of AD. These aims will be accomplished using the following technologies: fluorescence ration imaging of intracellular calcium levels, and patch clamp studies of calcium currents in hippocampal neurons; immunocytochemistry to localize beta-amyloid and calcium-regulating proteins in cell cultures and in AD brains; confocal laser scanning microscopy; electron microscopy. Taken together, these studies will: (1) Provide insight into the cellular and molecular mechanisms whereby beta-amyloid destabilizes neuronal calcium homeostasis. (2) Tell us whether the cellular pathology of AD is consistent, at the molecular level, with the calcium-destabilization hypothesis. (3) Generate information that can be used to develop new approaches to preventing and treating the neuronal damage that is responsible for the progression of AD.

这项研究的长期目标是了解 导致神经元损伤和死亡的细胞和分子事件 阿尔茨海默病（AD）。 该项目测试了以下假设：beta- 淀粉样蛋白破坏细胞钙稳态，从而导致 神经元更容易受到环境侵害。 海马细胞 培养系统将用于检查细胞和分子 β-淀粉样蛋白破坏神经元钙稳态的机制， 并增强兴奋性氨基酸神经毒性。 免疫定位 AD 大脑的研究旨在确定神经病理学是否 AD 的发生与 β-钙不稳定假说是一致的 淀粉样蛋白神经毒性。 第一个目标将检验以下假设：β- 淀粉样蛋白影响特定细胞系统的钙稳态（NMDA 受体、钙通道 Na+/Ca2+ 交换器、钙结合蛋白和 钙 ATP 酶）。 第二个目的是确定内源性β- 淀粉样蛋白有助于细胞培养中的选择性神经元脆弱性。 第三个目的是确定细胞骨架表现是否 β-淀粉样蛋白诱导的神经元变性类似于神经原纤维 AD 的病理学。 第四个目标是确定 beta- 的关系 淀粉样蛋白、NMDA 受体和钙调节蛋白 AD 的组织病理学。 这些目标将通过以下方式实现 技术：细胞内钙水平的荧光定量成像， 海马神经元钙电流的膜片钳研究； 免疫细胞化学定位 β-淀粉样蛋白和钙调节 细胞培养物和 AD 大脑中的蛋白质；共焦激光扫描 显微镜检查；电子显微镜。 总的来说，这些研究将：（1）深入了解细胞 以及β-淀粉样蛋白破坏神经元钙稳定的分子机制 体内平衡。 (2) 请告诉我们 AD 的细胞病理学是否是 在分子水平上与钙不稳定一致 假设。 (3) 生成可用于开发新产品的信息 预防和治疗神经元损伤的方法 AD 的进展负责。