Neuronal signal transduction in space and time using single quantum dots

使用单量子点进行空间和时间神经元信号转导

• 批准号: 8494700
• 负责人: Tothu Q Vu
• 金额: $ 32.43万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8494700
• 关键词: Address; Algorithms; Alzheimer' s Disease; Area; Behavior; Binding; Biochemical; Brain; Brain-Derived Neurotrophic Factor; Cause of Death; Cell membrane; Cells; Characteristics; Clinical; Complex; Cytoplasm; Data; Degenerative Disorder; Destinations; Development; Disease; Distant; Endocytosis; Endosomes; Event; Family; G-Protein-Coupled Receptors; Goals; Image; Imagery; Imaging Techniques; Imaging technology; Immunoblotting; Indium; Individual; Intracellular Transport; Investigation; Label; Learning; Life; Ligands; Location; MEKs; Maps; Masks; Memory; Molecular; Motion; Movement; Nerve Degeneration; Neuraxis; Neurodegenerative Disorders; Neurons; Neurotrophic Tyrosine Kinase Receptor Type 2; Parkinson Disease; Pathway interactions; Patients; Pattern; Pharmacologic Substance; Phosphorylation; Physiological; Play; Population Dynamics; Proteins; Psyche structure; Quantum Dots; Receptor Protein-Tyrosine Kinases; Receptor Signaling; Research; Research Personnel; Resolution; Role; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Signaling Molecule; Signaling Protein; Site; Societies; Surface; Synaptic plasticity; Techniques; Technology; Testing; Time; base; clinically relevant; disability; effective therapy; fluorophore; immunocytochemistry; improved; nanoparticle; nanoscale; neural growth; neuronal growth; novel; public health relevance; receptor; relating to nervous system; response; single molecule; spatial relationship; spatiotemporal; success; therapeutic target; time use; treatment strategy

DESCRIPTION (provided by applicant): The long-term goal of the proposed research is to understand how neurons transduce biochemical signals in space and time, at the molecular level. Brain-derived neurotrophic factor (BDNF) is highly expressed in the brain and activates critical receptor signaling pathways that dictate neuronal growth, synaptic plasticity, and memory. Decreased BDNF signaling is a key element in devastating neurodegenerative diseases, including Alzheimer's disease. Thus, BDNF signaling transduction pathways are attractive therapeutic targets. However, despite the important role of BDNF in the brain, mechanisms underlying BDNF signaling in the central nervous system are not well understood. Signaling complexes consisting of internalized BDNF receptors (BDNF-Rs) are hypothesized to represent a fundamental mechanism for propagating BDNF signaling. Unfortunately, understanding of these mechanisms- how BDNF-Rs move in space and time in neurons, and how BDNF-R spatiotemporal dynamics regulate downstream signaling events- remains poorly defined. We have recently shown that fluorescent nanoparticle quantum dots allow real-time, intracellular visualization of individual receptor complexes with nanoscale spatial resolution, thereby providing the first access to dynamic populations of individual BDNF- Rs previously invisible to more conventional imaging techniques. Accordingly, we propose to expand current single quantum dot (QD) imaging technologies to create novel, ultra-sensitive, and photostable QD probes capable of high-resolution imaging of the spatiotemporal behavior of single neuronal receptor complexes inside live cells. These capabilities will be applied to elucidate the spatiotemporal action of BDNF-R mechanisms in regulating downstream signaling pathways implicated in neurodegenerative diseases. We propose to develop new BDNF-QD probes and validate new algorithms for tracking and analyzing spatiotemporal BDNF signaling with single molecule sensitivity. We will: (1) identify the optimal monovalent QD bioconjugation strategy for physiological tracking of individual receptor signaling complexes within cells; (2) establish QD algorithms to track and analyze individual BDNF receptor complexes in neurons; (3) determine the role of BDNF-receptor complexes in propagating downstream cellular signaling. As BDNF-Rs belong to the family of tyrosine kinase receptors that, along with G-protein coupled receptors, make up 50% of all pharmaceutical targets, the technologies developed here will be relevant to other disease states in which impaired receptor signaling may play an important role.

描述（由申请人提供）：拟议的研究的长期目标是了解神经元如何在分子水平的时空中传递生化信号。 脑衍生的神经营养因子（BDNF）在大脑中高度表达，并激活关键的受体信号传导途径，该途径决定了神经元生长，突触可塑性和记忆力。 降低BDNF信号传导是包括阿尔茨海默氏病在内的毁灭性神经退行性疾病的关键因素。 因此，BDNF信号转导途径是有吸引力的治疗靶标。 然而，尽管BDNF在大脑中起着重要作用，但中枢神经系统中BDNF信号的基础机制尚不清楚。 假设由内部BDNF受体（BDNF-R）组成的信号传导复合物被认为代表了传播BDNF信号传导的基本机制。 不幸的是，了解这些机制 - BDNF-R如何在神经元中的空间和时间中移动，以及BDNF-R时空动力学如何调节下游信号事件 - 仍然很差。 我们最近显示，荧光纳米粒子量子点可以实时，具有纳米级空间分辨率的单个受体复合物的实时，细胞内可视化，从而为更传统的成像技术提供了先前不可见的单个BDNF-RS动态种群的首次访问。 因此，我们建议扩展当前的单量子点（QD）成像技术，以创建能够对活细胞中单个神经受体复合物的时空行为进行高分辨率成像的新颖，超敏感和光稳定的QD探针。 这些功能将用于阐明BDNF-R机制在调节与神经退行性疾病有关的下游信号通路中的时空作用。 我们建议开发新的BDNF-QD探针，并验证新算法，用于跟踪和分析具有单分子敏感性的时空BDNF信号传导。 我们将：（1）确定细胞内各个受体信号复合物的生理跟踪的最佳单价QD生物结合策略； （2）建立QD算法以跟踪和分析神经元中的单个BDNF受体复合物； （3）确定BDNF受体复合物在传播下游细胞信号传导中的作用。 由于BDNF-RS属于酪氨酸激酶受体家族，该家族与G蛋白偶联受体一起占所有药物靶标的50％，因此这里开发的技术将与其他疾病状态有关，在其他疾病状态下，受损受体信号可能起重要作用。