PUNQs: Photostable, Ultrafast, Nano-optode, Quantum-dots to image Na in dendrites

PUNQ：光稳定、超快、纳米光极、量子点，用于在树突中成像 Na

• 批准号: 8397274
• 负责人: Timothy Tordella Ruckh
• 金额: $ 5.22万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8397274
• 关键词: Acute; Acute Disease; Adolescent; Alzheimer' s Disease; Architecture; Axon; Binding; Biochemical; Biomedical Research; Brain; Brain Concussion; Cells; Charge; Chemistry; Chronic Disease; Dendrimers; Dendrites; Dendritic Spines; Development; Diffuse; Diffusion; Distal; Dyes; Electrophysiology (science); Elements; Event; Exhibits; Foundations; Future; Glucose; Glutamates; Goals; Head; Image; Individual; Injection of therapeutic agent; Interdisciplinary Study; Ions; Kinesin; Kinetics; Knowledge; Laboratories; Laser Scanning Microscopy; Learning; Length; Ligands; Measurement; Measures; Mediating; Memory; Methods; Microscopy; Modification; Morphology; Mus; Neck; Neuraxis; Neurobiology; Neurons; Pain; Parents; Peptides; Photobleaching; Photons; Physiological; Physiology; Play; Preparation; Process; Property; Quantum Dots; Reaction Time; Research; Research Personnel; Research Training; Role; Shapes; Signal Transduction; Slice; Sodium; Specificity; Spinal; Structure; Structure-Activity Relationship; Surface; Synapses; Temperature; Trauma; Vertebral column; chemical property; design; experience; fascinate; flexibility; hippocampal pyramidal neuron; insight; nano; nanoparticle; nanosensors; nervous system disorder; neural circuit; neuronal cell body; novel; painful neuropathy; patch clamp; research study; response; small molecule; time use; tool; two-photon

DESCRIPTION (provided by applicant): This research plan proposes a major technical advancement in fluorescent imaging of ion dynamics by designing modular, tunable nanosensors with narrow emission spectra. Under two-photon microscopy, these nanosensors will quantitatively image the spatio-temporal dynamics of sodium fluxes in dendritic spines, which has never been done before. Dendritic spines are tiny, semi-autonomously neuronal compartments that exhibit a fascinating relationship between their structure and their function in processing synaptic signals. This dynamic structure-function relationship provides extraordinarily high input specificity while also allowing for rapid modifications that give rise t plasticity in neural circuits. Understanding spinal physiology may vastly enhance our causal knowledge for a broad range of diseases from Alzheimer's to neuropathic pain as well as basic processes in learning and memory. In order to gain be able to image sodium fluxes, we need to design fluorescent nanosensors that can enter into spines and rapidly measure local sodium concentrations. For this reason, we will use a novel design we call PUNQs for Photostable, Ultra-fast, Nano-optode, Quantum dots. These new PUNQs are modular, tunable, and have narrow emission spectra. Thus, with existing chemistry, they can be easily modifiable to study new ion and small-molecule targets, their dynamic ranges can be adjusted for the target analyte's physiologic concentration, and they can be multiplexed together. This research will produce new insight into the elusive dynamics of sodium in dendritic spines, and the PUNQ platform will be applicable to any research involving ion dynamics. The specific aims of this research are: 1) Designing PUNQs with optimal size and chemical properties to achieve physiologically-relevant analyte sensitivity and selectivity 2) Delivering PUNQs into small cellular subcompartments at effective concentrations. 3) Measuring dendritic and somatic Na+ fluxes in response to glutamatergic excitation at dendritic spines. This interdisciplinary research merges the Clark laboratory's experience with nanosensor development, the Bhatia laboratory's expertise in multifunctional nanoparticle development, and the Sabatini laboratory's expertise in neurobiology. The proposed research and training plan will elucidate the role of sodium in dendritic spines, and provide a high-value, flexible tool to study ion dynamics within individual cells. Finally, research will provide me with valuable experience that will prepare me for a future as an independent investigator in biomedical research. PUBLIC HEALTH RELEVANCE: This research plan proposes a major technical advancement in fluorescent imaging of ion dynamics by designing modular, tunable nanosensors with narrow emission spectra. These nanosensors will be used to image sodium dynamics in dendritic spines, and they will be expanded to imaging multiple ions simultaneously in the future.

描述（由申请人提供）：本研究计划提出了通过设计具有狭窄发射光谱的模块化，可调的纳米传感器来实现离子动力学荧光成像的主要技术进步。在两光子显微镜下，这些纳米传感器将在树突棘中定量地对钠通量的时空动力学进行定量图像，这是以前从未做过的。树突状的棘是微小的半自主神经元室，其结构与它们在处理突触信号中的功能之间表现出迷人的关系。这种动态结构功能关系提供了极高的输入特异性，同时还允许快速修改可在神经回路中产生T可塑性。了解脊柱生理可能会极大地增强我们的因果知识，从阿尔茨海默氏症到神经性疼痛以及学习和记忆的基本过程。为了获得能力成像钠通量，我们需要设计荧光纳米传感器，这些荧光纳米传感器可以进入脊柱并迅速测量局部钠浓度。因此，我们将使用一种新颖的设计，我们称为PUNQ，用于光稳定，超快速，纳米式，量子点。这些新的PUNQ是模块化的，可调的，并且具有狭窄的发射光谱。因此，借助现有的化学，它们可以很容易地修改以研究新离子和小分子靶标，可以为目标分析物的生理浓度调整它们的动态范围，并且可以将它们一起多路复用。这项研究将对树突刺中钠难以捉摸的动态产生新的见解，PUNQ平台将适用于任何涉及离子动力学的研究。这项研究的具体目的是：1）设计具有最佳尺寸和化学特性的PUNQ，以实现与生理相关的分析物敏感性和选择性2）以有效的浓度将PUNQ交付到小细胞子店中。 3）测量树突状刺激的谷氨酸能激发的树突状和体细胞Na+通量。这项跨学科研究 将克拉克实验室的经验与纳米传感器开发，巴蒂亚实验室在多功能纳米颗粒开发方面的专业知识以及Sabatini实验室在神经生物学方面的专业知识相结合。拟议的研究和培训计划将阐明钠在树突状棘中的作用，并提供一种高价值的灵活工具来研究单个细胞内的离子动力学。最后，研究将为我提供宝贵的经验，这将使我为未来做好准备 作为生物医学研究的独立研究者。 公共卫生相关性：该研究计划提出了通过设计具有狭窄发射光谱的模块化，可调的纳米传感器来实现离子动力学荧光成像的主要技术进步。这些纳米传感器将用于图像树突棘中的钠动力学，并将将来将它们同时扩展为成像多个离子。