Imaging NGF signal transduction in live neurons

活神经元中 NGF 信号转导的成像

• 批准号: 7675579
• 负责人: Bianxiao Cui
• 金额: $ 25.11万
• 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 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7675579
• 关键词: Alzheimer' s Disease; Amyloid beta-Protein Precursor; Atrophic; Axon; Axonal Transport; Binding; Brain; C-terminal; Chromosomes, Human, Pair 16; Color; Complex; Defect; Down Syndrome; Endosomes; Energy Supply; Event; Fluorescence; Frequencies; Gene Expression; Green Fluorescent Proteins; Image; Imaging Techniques; Individual; Label; Lead; Length; Life; Link; Location; Maintenance; Measures; Microtubules; Mitochondria; Modeling; Molecular Motors; 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; Resolution; Role; Severities; Signal Pathway; Signal Transduction; Signaling Molecule; Specific qualifier value; Speed; System; Testing; Time; Travel; basal forebrain cholinergic neurons; cellular imaging; cholinergic neuron; fluorescence imaging; mouse Ts65Dn; nanometer; neuronal cell body; retrograde transport; single molecule; trafficking

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 forebarin 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 neruons 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 inlcude 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.

我们广泛的目标是了解神经生长因子（NGF）信号的机制 从轴突末端传播到细胞体。 NGF逆行信号对于生存至关重要， 某些类型神经元的分化和维护。报告了破坏的NGF逆行运输 有助于损失基底胆碱能（BFC）神经元患者的大脑 阿尔茨海默氏病或​​唐氏综合症。该项目将使用高级成像技术直接 实时可视化现场杂物中的NGF运输。我们专注于探索NGF的动态功能 正常综合征小鼠的运输。目的是： 1。表征轴突中含有NGF的内体的运动并定义其暂停 机理，通过使用量子点共轭NGF来跟踪纳米表的内体运动 解决。 2。确定是否存在NGF占用内体，是否与NGF信号相关， 以及是否存在独立于内体传输的替代信号通路，通过标记 具有光活化的绿色荧光蛋白，融合到C末端的NGF占用内体 TRKA受体。 3。通过 表征运输动力学的个体特征，该动力的平均速度，移动 速度，暂停持续时间和暂停频率。 4。确定淀粉样蛋白前体蛋白过表达如何导致异常NGF逆行 通过检查淀粉样蛋白在DS中的过度表达如何通过唐氏综合症小鼠的运输 小鼠可能会​​导致缺陷的结构或轴突特征，从而导致NGF转运破坏。 实现这些目标将增加我们对NGF信号在正常情况下如何传播的理解和 退化神经元。更广泛地，这些研究将有助于阐明 阿尔茨海默氏病和唐氏综合症。