CALBINDIN-D28K--ROLE IN NEURODEGENERATION

CALBINDIN-D28K——在神经退行性疾病中的作用

• 批准号: 2268394
• 负责人: DWIGHT C. German
• 金额: $ 29.7万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-08-01 至 1996-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2268394
• 关键词: Parkinson's disease; autoradiography; calcium binding protein; digital imaging; disease /disorder model; dopamine; enzyme activity; gene expression; genetic promoter element; genetically modified animals; immunocytochemistry; in situ hybridization; laboratory mouse; messenger RNA; methylphenyltetrahydropyridine; neural degeneration; phosphopyruvate hydratase; substantia nigra; transfection; tyrosine 3 monooxygenase

Calbindin-D28k (CaBP), a calcium-binding protein that is present in defined neuronal populations in the brain and spinal cord, appears to protect cells from calcium-mediated cellular degeneration. CaBP is found in the hippocampal neurons that are spared cell loss caused by experimentally induced seizures and excitotoxins. The neurons that do not contain CaBP appear to die as a result of toxic levels of intracellular Ca2+. Recent data from our laboratory indicate that the specific midbrain dopaminergic (DA) neurons that contain CaBP are preserved in patients with Parkinson's disease, and in monkeys and mice treated with 1-methyl- 4-phenyl-1,2,3,6-tetrahydropyridine (MPTP, a neurotoxin which produces a parkinsonian syndrome). However, those DA neurons that do not contain CaBP are most vulnerable to Parkinson's disease and MPTP. The purpose of the present grant is-to determine whether CaBP protects midbrain DA neurons from the neurotoxic effects of MPTP in the mouse. First, we will use immunohistochemical staining for CaBP, computer imaging, CaBP in situ hybridization, 14C-MPTP in vivo autoradiography, and monoamine biochemical measuring techniques to determine whether endogenous CaBP protects specific midbrain DA neurons from MPTP-induced degeneration. Secondly, we will develop animal models in which to test the protective effects of CaBP against MPTP by generating transgenic mice which express CaBP within all of the midbrain DA neurons. We will use the tyrosine hydroxylase promoter and the neuron specific enolase promoter to ex-press CaBP, and determine whether the presence of this protein will specifically protect MPTP-sensitive areas of the midbrain (substantia nigra). These experiments will: (I) determine whether CaBP protects midbrain DA neurons from MPTP-induced parkinsonism; and (2) provide animal models of other neurodegenerative diseases where calcium-mediated neurodegeneration may play a major etiologic role (e.g., Alzheimer's disease, Huntington's disease).

钙结合蛋白-D28k (CaBP)，一种钙结合蛋白，存在于 大脑和脊髓中确定的神经元群，似乎 保护细胞免受钙介导的细胞变性。发现CaBP 在海马神经元中，这些神经元免受由 实验诱发癫痫发作和兴奋性毒素。那些没有的神经元 含有 CaBP 的细胞似乎会因细胞内有毒水平而死亡 钙2+。我们实验室的最新数据表明，特定的中脑 患者体内保留了含有 CaBP 的多巴胺能 (DA) 神经元 帕金森病患者以及接受 1-甲基- 治疗的猴子和小鼠 4-苯基-1,2,3,6-四氢吡啶（MPTP，一种神经毒素，可产生 帕金森综合症）。然而，那些不包含的 DA 神经元 CaBP 最容易患帕金森病和 MPTP。目的 目前的资助是确定 CaBP 是否保护中脑 DA MPTP 对小鼠的神经毒性作用对神经元的影响。首先，我们将 使用免疫组织化学染色进行 CaBP、计算机成像、CaBP 原位 杂交、14C-MPTP 体内放射自显影和单胺 生化测量技术确定是否存在内源性 CaBP 保护特定的中脑 DA 神经元免受 MPTP 诱导的变性。 其次，我们将开发动物模型来测试保护性 CaBP 通过产生表达的转基因小鼠来对抗 MPTP 所有中脑 DA 神经元内的 CaBP。我们将使用酪氨酸 羟化酶启动子和神经元特异性烯醇化酶启动子表达 CaBP，并确定该蛋白质的存在是否会 专门保护中脑的 MPTP 敏感区域（实质 黑）。这些实验将： (I) 确定 CaBP 是否具有保护作用 MPTP 诱导的帕金森症的中脑 DA 神经元； (2) 提供 其他神经退行性疾病的动物模型，其中钙介导 神经退行性变可能发挥重要的病因作用（例如，阿尔茨海默病 病、亨廷顿病）。