Cholinergic neurons and memory in GFP-AD mouse: a novel neurotrophic therapy

GFP-AD 小鼠的胆碱能神经元和记忆：一种新型神经营养疗法

• 批准号: 8634611
• 负责人: ALPASLAN DEDEOGLU
• 金额: --
• 依托单位: VA BOSTON HEALTH CARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8634611
• 关键词: Acetylcholine; Adult; Affect; Age; Age-Months; Agonist; Alzheimer' s Disease; Alzheimer' s disease model; Amyloid; Amyloid beta-Protein Precursor; Amyotrophic Lateral Sclerosis; Animal Model; Animals; Asses; Attention; Axotomy; Biology; Blood - brain barrier anatomy; Brain; Brain-Derived Neurotrophic Factor; Cell Culture Techniques; Cells; Cerebral cortex; Cerebrum; Choline; Choline O-Acetyltransferase; Cholinergic Fibers; Code; Cognition; Cognitive; Cognitive deficits; Computer Assisted; Crossbreeding; Data; Dementia; Dendritic Spines; Densitometry; Development; Disease; Drug Kinetics; Elderly; Enzyme-Linked Immunosorbent Assay; Enzymes; Evaluation; Exhibits; Family Caregiver; Family member; Fluorescence-Activated Cell Sorting; Functional disorder; Future; Gene Expression; Gene Expression Profile; Genes; Genetic; Glutamine; Golgi Apparatus; Green Fluorescent Proteins; Healthcare Systems; High Pressure Liquid Chromatography; Hippocampus (Brain); Human; Immunohistochemistry; Individual; Inositol; Learning; Life; Magnetic Resonance Spectroscopy; Measures; Memory; Memory Loss; Memory impairment; Messenger RNA; Metabolic; Metabolism; Modeling; Mouse Strains; Mus; Mutate; Neocortex; Nerve Degeneration; Nerve Growth Factors; Neurofibrillary Tangles; Neurons; Neurotransmitters; Neurotrophic Tyrosine Kinase Receptor Type 2; Parkinson Disease; Pathology; Patients; Pattern; Peptides; Pharmacology; Population; Post-Traumatic Stress Disorders; Process; Production; Prosencephalon; Proteins; Psyche structure; Rattus; Reporting; Reverse Transcriptase Polymerase Chain Reaction; Role; Senile Plaques; Signal Transduction; Slice; Staining method; Stains; Synapses; Synaptophysin; System; Techniques; Testing; Therapeutic; Tissue Stains; Tissues; Transgenic Organisms; Veterans; Wild Type Mouse; age related; aged; amyloid pathology; basal forebrain; basal forebrain cholinergic neurons; brain tissue; cholinergic; cholinergic neuron; computerized; design; extracellular; familial Alzheimer disease; functional disability; gamma-Aminobutyric Acid; in vivo; indexing; mRNA Expression; morris water maze; mouse model; mutant; neurochemistry; neurogenesis; neurotrophic factor; novel; postnatal; presenilin-1; prevent; public health relevance; receptor; small molecule; tau Proteins; tau-1; trimethylamine

Alzheimer's disease (AD) is an age dependent neurodegenerative disorder and the most common form of dementia affecting the elderly. The neuropathological hallmarks of AD include extracellular ss-amyloid- containing plaques and intracellular neurofibrillary tangles composed of phosphorylated tau protein in the hippocampus and neocortex, and degeneration and loss of basal forebrain cholinergic neurons (BFCN). BFCN project to the hippocampus and cerebral cortex where the release of their neurotransmitter, acetylcholine (ACh), is central to cognition. A decline in BFCN function and cholinergic marker is apparent in AD patients and in animal models of AD. Thus, it has been postulated that the dysfunction and/or degeneration of BFCN contributes to the memory deficits associated with AD. Many of the available treatments for AD are designed to inhibit the breakdown of ACh, though the administration of trophic factors that support the function and survival of cholinergic neurons has also been explored. One such neurotrophin is brain derived neurotrophic factor (BDNF) that signals via the TrkB receptor. The reduced expression of mRNAs encoding TrkB in purified BFCN from AD patients, suggests that BDNF therapy could prevent or restore the functional impairments of these neurons in AD. This approach has been hampered by to the poor pharmacokinetics of BDNF that does not cross the blood-brain barrier. Recently a potent and selective TrkB agonist that readily enters the brain when administered peripherally was discovered 7,8-dihydroxyflavone (7,8-DHF). Beneficial effects of 7,8-DHF have been reported in models of Parkinson's disease, AD and post-traumatic stress disorder and we are presenting preliminary data of DHF treatment in mouse models of AD and amyotrophic lateral sclerosis that strongly support these reports. We hypothesize that 7,8-DHF, a TrkB agonist, will delay or lessen memory loss and reduce amyloid pathology through its neurotrophic effects on hippocampal and basal forebrain cholinergic neurons. To test our hypothesis APP.PS1/CHGFP mice will be treated with 7,8-DHF from 1 to 6 months (early pathology) or 12 months (late pathology). We will measure the effects of 7,8-DHF treatment on animals' memory and learning using Morris water maze test, BFCN gene expression pattern by microarray and RT-PCR of FACS purified BFCN, hippocampal ACh content and release by high-performance liquid chromatography (HPLC), amyloid pathology by immunohistochemistry (IHC) and ELISA and indices of hippocampal and cerebral cortical function assessed by IHC techniques including cholinergic neurons and cholinergic fibers, neurogenesis and synapses and dendritic spines. We will also measure neurochemical profile using magnetic resonance spectroscopy (MRS). Our proposed studies incorporate principals of rational pharmacology with state-of-the-art neuropathological, neurochemical, gene-analysis and MR techniques with cognitive evaluation to comprehensively assess the brain of a unique Alzheimer mouse model that expresses green fluorescent protein specifically in cholinergic cells. These studies will help further define the therapeutic benefits of a novel neurotrophic compound that crosses the blood-brain barrier but also will provide an entirely new level of understanding of the basic biology of BFCN and their function in AD and characterize a novel model for future use to test therapeutics relevant to patients with AD.

阿尔茨海默病(AD)是一种年龄相关的神经退行性疾病，也是最常见的形式 影响老年人的痴呆症。AD的神经病理特征包括细胞外SS-淀粉样蛋白- 含有斑块和由磷酸化tau蛋白组成的细胞内神经原纤维缠结 海马和新皮质，以及基底前脑胆碱能神经元的变性和丢失。BFCN 投射到海马体和大脑皮层，在那里它们的神经递质乙酰胆碱释放 (ACh)是认知的核心。AD患者BFCN功能和胆碱能标志物明显下降 在AD的动物模型中也是如此。因此，推测BFCN的功能障碍和/或退变 导致与AD相关的记忆缺陷。许多治疗阿尔茨海默病的现有治疗方法都旨在 抑制ACh的分解，尽管给予支持功能和生存的营养因子 对胆碱能神经元的研究也得到了探索。其中一种神经营养因子是脑源性神经营养因子。 (BDNF)通过TrkB受体传递信号。编码TrkB的mRNAs在纯化的BFCN中的表达降低 提示脑源性神经营养因子治疗可以预防或恢复这些神经元的功能损害。 AD中的神经元。由于BDNF的药代动力学不佳，这种方法受到了阻碍 穿过血脑屏障。最近，一种有效的选择性TrkB激动剂，当 外周给药发现7，8-二羟基黄酮(7，8-DHF)。7，8-DHF的有益作用有 在帕金森氏病、阿尔茨海默病和创伤后应激障碍的模型中都有报道，我们正在介绍 DHF治疗AD和肌萎缩侧索硬化症小鼠模型的初步数据 支持这些报告。我们假设TrkB激动剂7，8-DHF将延缓或减轻记忆丧失和 淀粉样蛋白对海马区和基底前脑胆碱能神经营养作用减轻病理改变 神经元。为了验证我们的假设，APP.PS1/CHGFP小鼠将在1到6个月(早期)接受7，8-DHF治疗 病理学)或12个月(晚期病理学)。我们将测量7，8-DHF处理对动物的影响 用Morris水迷宫测试记忆和学习，用基因芯片和RT-PCR检测BFCN基因表达谱 高效液相色谱法测定FACS纯化的BFCN、海马区ACh含量及释放 (高效液相色谱法)、免疫组织化学(IHC)和酶联免疫吸附试验(EL ISA)检测淀粉样蛋白病理变化，以及大鼠海马区和脑组织指数 用IHC技术评估大脑皮层功能，包括胆碱能神经元和胆碱能纤维， 神经发生、突触和树突。我们还将使用磁学技术测量神经化学特征 共振光谱学(MRS)。我们建议的研究将理性药理学的原理与 最新的神经病理学、神经化学、基因分析和磁共振技术及其认知评估 全面评估一种独特的表达绿色荧光的阿尔茨海默病小鼠模型的大脑 胆碱能细胞中特有的蛋白质。这些研究将有助于进一步确定一部小说的治疗益处 通过血脑屏障的神经营养化合物，但也将提供一个全新的水平 了解BFCN的基本生物学及其在阿尔茨海默病中的作用，并为未来的研究提供新的模型 用于测试与AD患者相关的治疗方法。