Single-Molecule Studies of Synaptic Vesicles

突触小泡的单分子研究

• 批准号: 8089428
• 负责人: Daniel T Chiu
• 金额: $ 32.73万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8089428
• 关键词: 3-phosphoglycerate; ATP Synthesis Pathway; Address; Affect; Amphetamines; Antibodies; Automobile Driving; Back; Binding; Biogenesis; Caliber; Calibration; Categories; Complex; Coupled; Coupling; Crowding; Diffusion; Dimensions; Environment; Enzymes; Exocytosis; Expenditure; Extravasation; Floor; Fluorescence; Fluorescence Microscopy; Fluorescent Antibody Technique; Gel; Glass; Glutamate Transporter; Glutamates; Glyceraldehyde 3-Phosphate; Hand; Hypoglycemia; Individual; Intake; Integral Membrane Protein; Investigation; Kinetics; Knowledge; Label; Lasers; Learning; Length; Lipid Bilayers; Lipids; Location; Macromolecular Complexes; Maps; Measurement; Measures; Membrane; Membrane Fusion; Membrane Proteins; Microscope; Modeling; Molecular; Monitor; Motion; Movement; Nature; Neurologic; Neurotransmitters; Nigericin; Oregon; Organelles; Oxidoreductase; Pharmaceutical Preparations; Phosphoglycerate Kinase; Physiology; Play; Positioning Attribute; Presynaptic Terminals; Procedures; Process; Protein Isoforms; Proteins; Proteomics; Proton Pump; Proton-Translocating ATPases; Pump; Recycling; Relative (related person); Reporting; Reserpine; Resolution; Role; Rose Bengal; Rotation; Running; SNAP receptor; Sampling; Site; Solutions; Sorting - Cell Movement; Spatial Distribution; Staging; Study Section; Surface; Synapses; Synaptic Cleft; Synaptic Receptors; Synaptic Transmission; Synaptic Vesicles; Synaptic plasticity; Synaptophysin; System; Techniques; Testing; Thick; Thinking; Time; Transgenic Mice; Transmembrane Domain; Trypan Blue; Valinomycin; Vesicle; Viscosity; Water; Work; base; dimer; driving force; gamma-Aminobutyric Acid; inhibitor/antagonist; inorganic phosphate; insight; interest; monoamine; nervous system disorder; neurotransmission; neurotransmitter release; pH gradient; postsynaptic; presynaptic; prevent; protein complex; protein function; receptor; research study; response; sensor; single molecule; small molecule; statistics; stoichiometry; synaptic function; synaptogyrin; synaptotagmin; trafficking; uptake

DESCRIPTION (provided by applicant): Gaining detailed knowledge on the workings of the vesicular transmitter uptake machinery is important in understanding synaptic function and plasticity, because this uptake process governs directly the number of transmitters released into the synaptic cleft and thus the degree of activation of the postsynaptic receptors. Determining the mechanism and rates of vesicular transmitter uptake also is important towards understanding the kinetics of vesicle recycling at the synapse and the energy requirements for loading and maintaining the filled vesicle at the presynaptic terminal. Given synaptic transmission is dependent on the uptake and storage of neurotransmitters within vesicles, it is not surprising that they are sites of action for many drugs, and thus understanding the functioning of this important molecular machinery will also have important pharmacological implications. Our proposed single-molecule and single-vesicle experiments are targeted towards deciphering the molecular organization and mechanism by which neurotransmitters are loaded into the synaptic vesicle. Towards this end, our specific aims are: Aim 1: Rotational measurements of synaptic vesicle integral membrane proteins and of small molecules contained within the vesicle Aim 2: Single-molecule studies of the vesicular H+ATPase Aim 3: Single-molecule studies of the vesicular glutamate transporters Aim 4: Investigation of the coupling of transmitter uptake to the glycolytic enzymes on synaptic vesicles and the spatial organization of the uptake machinery. From these experiments, we will develop an in-depth understanding of the workings of this complex molecular machinery with single-molecule resolutions, and offer new insight into how malfunctioning caused by neurological diseases or targeted disruption of this machinery with drugs can affect synaptic transmission.

描述（由申请人提供）：获得有关囊泡发射器摄取机制的工作的详细知识，对于理解突触功能和可塑性很重要，因为这种摄取过程直接控制了释放到突触left中的发射机数量，因此在突触后受体的激活程度。确定囊泡发射器摄取的机理和速率对于理解突触时囊泡回收的动力学以及在突触前末端加载和维持填充囊泡的能量需求也很重要。给定突触传播取决于神经递质在囊泡中的摄取和储存，毫不奇怪，它们是许多药物的作用部位，因此了解这种重要的分子机械的功能也将具有重要的药理意义。我们提出的单分子和单维体实验旨在解密分子组织和机制，并将神经递质加载到突触囊泡中。 Towards this end, our specific aims are: Aim 1: Rotational measurements of synaptic vesicle integral membrane proteins and of small molecules contained within the vesicle Aim 2: Single-molecule studies of the vesicular H+ATPase Aim 3: Single-molecule studies of the vesicular glutamate transporters Aim 4: Investigation of the coupling of transmitter uptake to the glycolytic enzymes在突触囊泡和摄取机械的空间组织上。从这些实验中，我们将深入了解通过单分子分辨率的这种复杂分子机械的起作用，并提供有关神经系统疾病引起的故障或对这种机械的靶向破坏如何影响突触传播的新见解。

项目成果

Ratiometric temperature sensing with semiconducting polymer dots.

• DOI: 10.1021/ja202945g
• 发表时间: 2011-06-01
• 期刊: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
• 影响因子: 15
• 作者: Ye, Fangmao;Wu, Changfeng;Jin, Yuhui;Chan, Yang-Hsiang;Zhang, Xuanjun;Chiu, Daniel T.
• 通讯作者: Chiu, Daniel T.

Probing the interior of synaptic vesicles with internalized nanoparticles.

用内化纳米颗粒探测突触小泡的内部。

• DOI: 10.1117/12.905091
• 发表时间: 2012
• 期刊: Proceedings of SPIE--the International Society for Optical Engineering
• 作者: Gadd,JenniferC;Budzinski,KristiL;Chan,Yang-Hsiang;Ye,Fangmao;Chiu,DanielT
• 通讯作者: Chiu,DanielT

Hybrid semiconducting polymer dot-quantum dot with narrow-band emission, near-infrared fluorescence, and high brightness.

• DOI: 10.1021/ja3022973
• 发表时间: 2012-05-02
• 期刊: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
• 影响因子: 15
• 作者: Chan, Yang-Hsiang;Ye, Fangmao;Gallina, Maria Elena;Zhang, Xuanjun;Jin, Yuhui;Wu, I-Che;Chiu, Daniel T.
• 通讯作者: Chiu, Daniel T.

Stable functionalization of small semiconducting polymer dots via covalent cross-linking and their application for specific cellular imaging.

• DOI: 10.1002/adma.201201245
• 发表时间: 2012-07-10
• 期刊: ADVANCED MATERIALS
• 影响因子: 29.4
• 作者: Yu, Jiangbo;Wu, Changfeng;Zhang, Xuanjun;Ye, Fangmao;Gallina, Maria Elena;Rong, Yu;Wu, I-Che;Sun, Wei;Chan, Yang-Hsiang;Chiu, Daniel T.
• 通讯作者: Chiu, Daniel T.

Development of ultrabright semiconducting polymer dots for ratiometric pH sensing.

• DOI: 10.1021/ac103140x
• 发表时间: 2011-02-15
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Chan, Yang-Hsiang;Wu, Changfeng;Ye, Fangmao;Jin, Yuhui;Smith, Polina B.;Chiu, Daniel T.
• 通讯作者: Chiu, Daniel T.