ACHE, CHAT & CHOLINERGIC NEURONS IN AGING & AD

疼痛，聊天

• 批准号: 3117788
• 负责人: STEVEN G YOUNKIN
• 金额: $ 21.22万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-12-01 至 1994-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3117788
• 关键词: Alzheimer's disease; PC12 cells; RNA splicing; aging; amyloid proteins; antiserum; brain cell; carboxyl group; cerebrospinal fluid; chemical cleavage; choline acetyltransferase; chromatography; electrophoresis; gene expression; human tissue; immunocytochemistry; laboratory rabbit; membrane proteins; messenger RNA; neurochemistry; nucleic acid sequence; parasympathetic nervous system; protease inhibitor; protein sequence; proteins; proteolysis; purine /pyrimidine metabolism; scintillation counter; solubility; tissue /cell culture; transfection; western blottings

In Alzheimer's disease (AD), there are striking changes in the distribution of choline acetyltransferase (ChAT) and the various molecular forms of acetylcholinesterase (AChE) in the cerebral cortex and nucleus basalis of Meynert (nbM). Our provisional working hypothesis is that these changes are due primarily to altered axonal transport in diseased cholinergic neurons. The major emphasis is on the acquisition of secure information on ChAT, AChE, and the corresponding mRNAs within individual cholinergic neurons and on the development of a system capable of generating similar information on many other proteins and mRNAs. We propose to evaluate the effect of aging and AD on ChAT and the varius molecular forms of AChE (a) on a per neuron basis in cholinergic nbM neurons, (b) at stereotyped locations along the course of cholinergic axons projecting from the nbM, (c) on a per axon basis in cholinergic axons of the fornix, and (d) in cerebral cortex and hippocampus. The effect of aging and AD on ChAT are AChE mRNA in cholinergic nbM neurons will also be evaluated on a per neuron basis. If normal aging causes changes qualitatively similar to those in AD, one may be able to gain considerable insight into the initiating events and the progression of the neuronal pathology in AD by carefully examining non-demented individuals in appropriate age groups. It is primarily for this reason that examination of the effect of aging is proposed. We plan to evaluate the effect of the many drugs that alter fast and/or slow axonal transport on ChAT and the various molecular forms of AChE in cultured PC12 cells. The goal is (a) to support our working hypothesis that impaired axonal transport plays an important role in the neuronal pathology of AD by demonstrating that appropriate impairment of axonal transport causes changed in ChAT and AChE like those observed in AD, and (b) to determine whether changes like those seen in AD occur only after specific perturbations of transport or develop non-specifically. The altered distribution of ChAT and AChE caused by axonal transport inhibitors could result from associated changes in any of the processes that regulate the level of these proteins. To assess this issue, the synthesis, assembly, secretion, and degradation of the various molecular forms of AChE will be assessed in drug-treated and control PC12 cells using techniques previously developed in this laboratory.

在阿尔茨海默病 (AD) 中，分布发生显着变化 胆碱乙酰转移酶 (ChAT) 及其各种分子形式 大脑皮层和基底核中的乙酰胆碱酯酶（AChE） 梅纳特（nbM）。 我们的临时工作假设是这些变化 主要是由于患病胆碱能神经元轴突运输的改变 神经元。 主要重点是获取有关的安全信息 ChAT、AChE 和个体胆碱能内相应的 mRNA 神经元以及能够产生类似神经元的系统的开发 许多其他蛋白质和 mRNA 的信息。 我们建议评估 衰老和 AD 对 ChAT 和 AChE 各种分子形式的影响 (a) 在胆碱能 nbM 神经元的每个神经元的基础上，(b) 在定型 沿着从 nbM 突出的胆碱能轴突的位置， (c) 以穹窿胆碱能轴突为基础，以及 (d) 大脑皮层和海马体。 衰老和 AD 对 ChAT 的影响是 胆碱能 nbM 神经元中的 AChE mRNA 也将在每个神经元上进行评估 基础。 如果正常衰老引起的质变类似于 AD，人们也许能够对始发事件有深入的了解 通过仔细检查 AD 神经元病理学的进展 适当年龄组的非痴呆个体。 它主要是为了 因此建议检查老化的影响。 我们计划 评估许多改变快速和/或缓慢轴突的药物的效果 培养的 PC12 中 ChAT 和 AChE 的各种分子形式的转运 细胞。 目标是（a）支持我们的工作假设： 轴突运输在 AD 的神经元病理学中起着重要作用 证明轴突运输的适当损伤会导致 ChAT 和 AChE 的变化与 AD 中观察到的类似，并且 (b) 确定 是否像 AD 中看到的那样的变化只有在特定的时间之后才会发生？ 运输扰动或非特异性发展。 改变后的 轴突转运抑制剂引起的 ChAT 和 AChE 分布可能 任何调节流程的相关变化所导致的 这些蛋白质的水平。 为了评估这个问题，合成、组装、 各种分子形式的 AChE 的分泌和降解将 使用先前的技术在药物处理和对照 PC12 细胞中进行评估 是在这个实验室开发的。