EXCITATORY AA RECEPTORS' IN ALZHEIMER'S AND OTHER NEURODEGENERATIVE DISORDERS

阿尔茨海默病和其他神经退行性疾病中的兴奋性 AA 受体

• 批准号: 6160455
• 负责人: J W KUSIAK
• 金额: --
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6160455
• 关键词: Alzheimer's disease; NMDA receptors; aging; biological signal transduction; excitatory aminoacid; fibroblast growth factor; genetic promoter element; glutamate receptor; glutamates; laboratory mouse; molecular cloning; neural degeneration; neurotrophic factors; receptor expression; tissue /cell culture

Summary of work: Glutamate receptors play a pivotal role in several brain functions including fast excitatory neurotransmission, induction and maintenance of long term potentiation, and synaptic plasticity. However, over-activation of these receptors is thought to initiate a common final pathway of neuronal cell death in both acute and chronic brain insults. A major focus of this project is to discover the pathological roles that excitatory amino acid (glutamate) receptors play in neuronal cell loss in aging and Alzheimer's Disease, and the mechanisms by which this cell loss occurs. One of our objectives is to determine how the N-methyl-D-Aspartate 1 receptor gene (NMDAR1) and other family member genes are regulated at the transcriptional level. Another objective of this project is to determine the mechanism by which glutamate causes cell death and the role activation of glutamate receptors plays in initiating a genetic cascade of programmed cell death. The promoter region of the NMDAR1 contains several transcriptional elements in the proximal region responsible for both basal, inducible, and neuronal specific expression. In recent work, we showed that nerve growth factor and other neurotrophins were able to stimulate expression of NMDAR1 promoter-Luciferase reporter constructs in PC12 cells. This stimulation was induced through activation of high affinity TrkA receptors and subsequent activation of a Ras/Raf MAP kinase pathway. We showed by electro-mobility shift assays that this activation may be mediated through both GSG and Sp1 transcription factor activation. Other elements in the NMDAR1 promoter are being studied including two Cyclic AMP responsive consensus sites in the proximal region. Also, we showed that glutamate treatment of a hippocampal cell line caused cell death which could be prevented by the over-expression of Bcl-2 protein. Since Bcl-2 can protect and we see increased amounts of DNA laddering in glutamate treated cells, the results suggest that this line may be a good model to study neuronal apoptosis. These cells express AMPA and kainate subtype glutamate receptors. NT2-N cells, on the other hand express the NMDA subtype of receptor and may be a good model of glutamate-induced necrotic cell death.

工作摘要：谷氨酸受体在几个大脑中起关键作用 功能包括快速兴奋性神经传递，诱导和 维持长期增强和突触可塑性。然而， 人们认为这些受体过度激活这些受体会引发共同的最终最终 急性和慢性大脑侮辱中神经元细胞死亡的途径。一个 该项目的主要重点是发现病理角色 兴奋性氨基酸（谷氨酸）受体在神经元细胞损失中发挥作用 衰老和阿尔茨海默氏病，以及这种细胞损失的机制 发生。我们的目标之一是确定N-甲基-D-天冬氨酸如何 1受体基因（NMDAR1）和其他家庭成员基因受调节 转录级别。该项目的另一个目标是 确定谷氨酸导致细胞死亡和作用的机制 谷氨酸受体的激活在启动遗传级联 程序性细胞死亡。 NMDAR1的启动子区域包含几个转录 近端区域的元素负责基础，诱导和 神经元特异性表达。在最近的工作中，我们证明了神经成长 因子和其他神经营养蛋白能够刺激NMDAR1的表达 PC12细胞中的启动子 - 卢西斯酶报告基因构建体。这种刺激 通过激活高亲和力TRKA受体和 随后的RAS/RAF MAP激酶途径的激活。我们展示了 电动移动转移测定法可能会通过 GSG和SP1转录因子激活。其他元素 正在研究NMDAR1启动子，其中包括两个环状AMP响应 近端区域的共识位点。另外，我们证明了谷氨酸 海马细胞系的治疗导致细胞死亡，这可能是 Bcl-2蛋白的过表达阻止。因为bcl-2可以保护 而且我们看到在谷氨酸处理的细胞中增加了增加的DNA梯子， 结果表明，这条线可能是研究神经元的好模型 凋亡。这些细胞表达AMPA和海藻酸盐亚型谷氨酸 受体。另一方面，NT2-N细胞表示NMDA亚型 受体，可能是谷氨酸诱导的坏死细胞死亡的良好模型。