CaMKI, Apoptosis and Cholinergic Neurons in Aging and AD

衰老和 AD 中的 CaMKI、细胞凋亡和胆碱能神经元

• 批准号: 7284208
• 负责人: CHANGIZ GEULA
• 金额: $ 6.45万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-15 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7284208
• 关键词: Aging; Alzheimer' s Disease; Apoptosis; Apoptotic; Attention; Axon; Basal Nucleus of Meynert; Brain; Ca(2+)-Calmodulin Dependent Protein Kinase; Calcium; Calcium-Binding Proteins; Cells; Cerebral cortex; Cholinergic Agents; Cyclic AMP-Responsive DNA-Binding Protein; Down-Regulation; Elderly; Future; Gene Expression; Genes; Human; Laboratories; Lasers; Lead; Learning; Literature; Memory; Methodology; Microscopy; Mus; Neurodegenerative Disorders; Neurons; Polymerase Chain Reaction; Process; Prosencephalon; Proteins; RNA; Reporting; Rodent; System; Testing; Time; Up-Regulation; adeno-associated viral vector; age related; basal forebrain; basal forebrain cholinergic neurons; base; calbindin-D28K; caspase-10; cholinergic; cholinergic neuron; immunoreactivity; nerve supply; nervous system disorder; neurochemistry; normal aging; novel; research study

DESCRIPTION (provided by applicant): The basal forebrain cholinergic neuronal (BFCN) system is intimately involved in the processes of learning, memory and attention. Within the human brain, the BFCN are selectively vulnerable in neurodegenerative diseases that afflict the elderly, such as Alzheimer's disease (AD). Recent preliminary observations from our laboratory indicate that the BFCN in AD display decreased expression of the gene for calmodulin kinase I (CaMKI) and increased immunoreactivity for the apoptotic protein Fas-associated death domain (FADD). It has been shown that CaMKI downregulation causes apoptosis in neurons. Thus, the downregulation of CaMKI expression in AD BFCN may be associated with apoptosis in these neurons. We have also observed that the BFCN display a substantial loss of the calcium binding protein calbindin-D28K (CB) in the course of normal aging in the human and a further decrease in Alzheimer's disease and that the BFCN which degenerate in Alzheimer's disease appear to be those which lose this protein in the course of normal aging. We hypothesize that downregulation of CaMKI in aging and AD BFCN results in upregulation of pro-apoptotic and / or downregulation of anti- apoptotic genes, and that changes in the expression of the above genes may be related to the age-related loss of CB, which is the earliest change observed in these neurons and may result in calcium dysregulation. The experiments in this proposal are intended to expand the scope of our previous studies using more novel methodologies. The proposed experiments will test the hypotheses that using single cell laser capture microscopy combined with quantitative real time PCR analysis : A. The BFCN in aging and Alzheimer's disease will display reduced expression of genes for CaMKI and the cyclic AMP response element binding protein (CREB), increased expression of pro-apoptotic genes and / or decreased expression of anti-apoptotic genes; B. altered expression of the above genes will occur primarily in CB-negative BFCN; C. decreased expression of CaMKI in cultured mouse BFCN using small inhibitory RNA will result in similar alterations in apoptosis- related genes; and D. increased expression of CB using adeno-associated viral vectors will result in upregulation of CaMKI. The results will have important implications for selective depletion of BFCN in aging and neurological diseases. They will also form the basis of more comprehensive future studies aimed at determining the causes and effects of the alterations of the above genes.

描述（由申请人提供）：基底前脑胆碱能神经元（BFCN）系统与学习、记忆和注意力过程密切相关。在人脑中，BFCN 在困扰老年人的神经退行性疾病（如阿尔茨海默氏病 (AD)）中选择性地受到影响。我们实验室最近的初步观察表明，AD 中的 BFCN 钙调蛋白激酶 I (CaMKI) 基因表达降低，凋亡蛋白 Fas 相关死亡结构域 (FADD) 免疫反应性增加。研究表明，CaMKI 下调会导致神经元凋亡。因此，AD BFCN 中 CaMKI 表达的下调可能与这些神经元的凋亡有关。我们还观察到，在人类正常衰老过程中，BFCN 表现出钙结合蛋白钙结合蛋白-D28K (CB) 的大量损失，并且在阿尔茨海默病中进一步减少，并且在阿尔茨海默病中退化的 BFCN 似乎是在正常衰老过程中失去这种蛋白质的 BFCN。我们假设衰老和 AD BFCN 中 CaMKI 的下调导致促凋亡基因的上调和/或抗凋亡基因的下调，并且上述基因表达的变化可能与年龄相关的 CB 损失有关，这是在这些神经元中观察到的最早的变化，并可能导致钙失调。本提案中的实验旨在使用更新颖的方法来扩大我们之前的研究范围。拟议的实验将测试使用单细胞激光捕获显微镜结合定量实时 PCR 分析的假设： A. 衰老和阿尔茨海默病中的 BFCN 将显示 CaMKI 和环 AMP 反应元件结合蛋白 (CREB) 基因表达减少，促凋亡基因表达增加和/或抗凋亡基因表达减少； B. 上述基因的表达改变主要发生在CB阴性的BFCN中； C.使用小抑制性RNA降低培养的小鼠BFCN中CaMKI的表达将导致凋亡相关基因的类似改变； D. 使用腺相关病毒载体增加 CB 的表达将导致 CaMKI 的上调。该结果将对衰老和神经系统疾病中 BFCN 的选择性消耗具有重要意义。它们还将构成未来更全面研究的基础，旨在确定上述基因改变的原因和影响。