Synaptic Proteins, Trophic Factors and Neurodegeneration

突触蛋白、营养因子和神经变性

• 批准号: 6665235
• 负责人: Gloria E. Meredith
• 金额: $ 28.68万
• 依托单位: ROSALIND FRANKLIN UNIV OF MEDICINE & SCI
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-05-01 至 2005-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6665235
• 关键词: Lewy body; Parkinson's disease; alpha synuclein; brain derived neurotrophic factor; cell death; electron microscopy; fluorescence microscopy; image processing; immunocytochemistry; in situ hybridization; laboratory mouse; methylphenyltetrahydropyridine; neural degeneration; neurochemistry; neurons; polymerase chain reaction; protein structure function; statistics /biometry; substantia nigra; synapses

Description (Provided by applicant): One of the most fundamental questions related to the progressive nature of neurodegeneration in human disease is how neurons die. Protecting nerve cells against morphological decline and death requires blocking intrinsic factors that inhibit neural repair. In the present proposal, we offer an innovative approach to study those factors that are active in Parkinson's disease (PD) in a new mouse model that shows synaptic loss and irreversible nigrostriatal degeneration. We propose to track changes of a key synaptic protein, a-synuclein, both in its native environment at presynaptic terminals and under neurotoxic conditions, when it becomes insoluble and accumulates. We will further correlate those changes with altered neurotrophic support. We have established an animal protocol by treating C57/bl mice with a combined regimen of 10 doses of probenecid at 250mg/kg and MPTP at 25mg/kg for 5 weeks. These mice show a slow, progressive loss of nigrostriatal dopaminergic function for at least 6 months, that mimics PD, with no signs of recovery. Three weeks after drug treatment, there is a significant reduction in the number of substantia nigra (SN) cells and dramatic changes in the subsynaptic distribution and density of a-synuclein-immunoreactive terminals. These changes could signal the beginning of a chain of events that leads to cell death. In this proposal, we will focus on the progressive deterioration of dopaminergic neurons in the SN and their inputs, and present three specific aims to be addressed through a series of hypotheses. Specifically, we plan to 1) ascertain the origin and neurochemical phenotype of synapses in the SN that contain a-synuclein and to establish whether MPTP + probenecid treatment leads to their degeneration; 2) determine, in the MPTP+P model, the temporal relationships between cell death and a-synuclein-positive synapses, decline in dopamine function and behavior; and 3) ascertain whether changes in a-synuclein expression and production are precipitated by altered neurotrophic support. The overall objective of our research is to understand the relationship between the synaptic protein, a-synuclein, neurotrophic support, especially brain-derived neurotrophic factor (BDNF) and their respective roles in the PD form of neurodegeneration. The findings of this research should shed light on target areas where neuroprotection strategies can be implemented.

描述（由申请人提供）：最基本的问题之一 与人类疾病中神经变性的进行性相关的是 神经元死亡。保护神经细胞免受形态衰退和死亡 需要阻断抑制神经修复的内在因素。在现在 提案中，我们提供了一种创新的方法来研究那些因素 在一种显示突触的新小鼠模型中，它对帕金森病 (PD) 具有活性 损失和不可逆的黑质纹状体变性。我们建议跟踪更改 一个关键的突触蛋白，a-突触核蛋白，都在其天然环境中 突触前末梢和神经毒性条件下，当它变​​成 不溶解并积累。我们将进一步将这些变化与改变的 神经营养支持。我们通过治疗 C57/bl 建立了动物方案 小鼠接受 10 剂丙磺舒（250mg/kg）和 MPTP（250mg/kg）的联合治疗方案 25毫克/公斤，持续5周。这些小鼠表现出黑质纹状体缓慢、进行性丧失 多巴胺能功能持续至少 6 个月，类似于 PD，没有任何迹象 恢复。药物治疗三周后， 黑质 (SN) 细胞的数量和 α-突触核蛋白免疫反应末端的突触亚分布和密度。 这些变化可能标志着一系列事件的开始，从而导致 细胞死亡。在本提案中，我们将重点关注日益恶化的情况 SN 中的多巴胺能神经元及其输入，并提出了三种特定的 目的是通过一系列假设来解决。具体来说，我们计划 1）确定SN中突触的起源和神经化学表型 含有α-突触核蛋白并确定 MPTP + 丙磺舒治疗是否有效 导致他们的退化； 2）确定MPTP+P模型中的时间 细胞死亡与α-突触核蛋白阳性突触之间的关系， 多巴胺功能和行为； 3) 确定α-突触核蛋白是否发生变化 神经营养支持的改变促进了表达和产生。这 我们研究的总体目标是了解 突触蛋白、α-突触核蛋白、神经营养支持，尤其是脑源性的 神经营养因子 (BDNF) 及其在 PD 形式中各自的作用 神经变性。这项研究的结果应该有助于明确目标 可以实施神经保护策略的领域。