Imaging nerve growth factor signal transduction in live neurons

活神经元中神经生长因子信号转导的成像

• 批准号: 7223656
• 负责人: Bianxiao Cui
• 金额: $ 5.75万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-12-01 至 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7223656
• 关键词: Alzheimer' s Disease; Amyloid beta-Protein Precursor; Atrophic; Axon; Axonal Transport; Binding; Brain; C-terminal; Cells; Cholinergic Agents; Chromosomes, Human, Pair 16; Color; Defect; Down Syndrome; Endosomes; Energy Supply; Event; Fluorescence; Frequencies; Genes; Green Fluorescent Proteins; Image; Imaging Techniques; Individual; Label; Lead; Length; Life; Link; Localized; Location; Maintenance; Measures; Microtubules; Mitochondria; Modeling; Motor; Movement; Mus; Nerve Growth Factors; Neurons; Neurotrophic Tyrosine Kinase Receptor Type 1; Numbers; Pathogenesis; Patients; Pattern; Presynaptic Terminals; Protein Overexpression; Proteins; Quantum Dots; Rate; Reporting; Research Personnel; Resolution; Role; Severities; Signal Pathway; Signal Transduction; Specific qualifier value; Speed; Syndrome; System; Testing; Time; Travel; basal forebrain; basal forebrain cholinergic neurons; cellular imaging; cholinergic; fluorescence imaging; mouse Ts65Dn; nanometer; neuronal cell body; programs; retrograde transport; single molecule; trafficking

DESCRIPTION (provided by applicant): Our broad objective is to understand the mechanisms by which the nerve growth factor (NGF) signal is propagated from the axon terminal to the cell body. NGF retrograde signaling is critical for the survival, differentiation, and maintenance of certain types neurons. Disrupted NGF retrograde transport was reported to contribute to the loss of the basal forebrain cholinergic (BFC) neurons in the brains of patients with Alzheimer's Disease or Down's Syndrome. This project will use advanced imaging techniques to directly visualize NGF transport in live neurons in real time. We focus on exploring dynamic features of NGF transport in normal and Down's Syndrome mice. The aims are: 1. Characterize the movement of NGF-containing endosomes in axons and define their pausing mechanism(s), by using quantum dot conjugated NGF to track endosomal movements with nanometer resolution. 2. Determine whether NGF-lacking endosomes are present, whether they are relevant for NGF signaling, and whether there are alternative signaling pathways independent of endosomal transport, by marking the NGF-lacking endosomes with photo-activatable green fluorescence proteins that are fused to the C-terminal of TrkA receptor. 3. Identify the abnormal features of disrupted NGF transport in Down Syndrome mouse neurons, by characterizing individual features of transport dynamics, which include the average speed, the moving speed, the pausing duration, and the pausing frequency. 4. Determine how amyloid precursor protein overexpression leads to the abnormal NGF retrograde transport in Down Syndrome mouse by examining how overexpression of amyloid precursor protein in DS mice might cause defective structural or axonal features that lead to disrupted NGF transport. Achieving those aims will increase our understanding of how NGF signal is propagated in normal and degenerative neurons. More broadly, those studies will contribute to elucidate the pathogenesis of Alzheimer's disease and Down syndrome.

描述（由申请人提供）： 我们的广泛目标是了解神经生长因子（NGF）信号从轴突末端传播到细胞体的机制。 NGF逆行信号对于某些类型的神经元的生存，分化和维持至关重要。据报道，NGF逆行转运破坏导致阿尔茨海默氏病或​​唐氏综合症患者的大脑中基底前脑胆碱能（BFC）神经元的丧失。该项目将使用先进的成像技术直接实时可视化活神经元中的NGF传输。我们专注于探索正常和唐氏综合症小鼠中NGF传输的动态特征。目的是：1。表征轴突中含有NGF的内体的运动，并通过使用量子点共轭NGF来以纳米分辨率跟踪内体运动，从而定义其暂停机制。 2。确定是否存在NGF占用的内体，是否与NGF信号传导相关，以及是否存在与内体传输无关的替代信号通路，通过标记具有可照相的绿色荧光蛋白的NGF占用型内体，它们与TRKA受体C-末端融合在一起。 3。通过表征传输动力学的各个特征，包括平均速度，移动速度，暂停持续时间和暂停频率，确定唐氏综合症小鼠神经元中NGF转运的异常特征。 4.通过检查淀粉样蛋白的过表达如何导致唐氏综合征小鼠中异常NGF逆行转运，通过研究ds小鼠中淀粉样蛋白前体蛋白的过表达如何导致缺陷的结构或轴突特征，从而导致NGF转运中断。实现这些目标将增加我们对正常和退化神经元中NGF信号如何传播的理解。更广泛地说，这些研究将有助于阐明阿尔茨海默氏病和唐氏综合症的发病机理。