Dynamic mapping of the complete synaptome using recombinant probes

使用重组探针动态绘制完整突触组

• 批准号: 8754412
• 负责人: DONALD B ARNOLD
• 金额: $ 188.59万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-25 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8754412
• 关键词: Address; Animal Testing; Animals; Behavior; Behavior Control; Benchmarking; Big Data; Binding; Brain; Brain Mapping; Cataloging; Catalogs; Cells; Cognition; Complement; Complex; Computer software; DLG4 gene; Data; Data Analyses; Development; Disease; Electrodes; Excitatory Synapse; Fibronectins; Floods; Fluorescence; Glutamates; Goals; Grant; Image; Image Analysis; Inhibitory Synapse; Knowledge; Label; Larva; Learning; Life; Light; Location; Maps; Memory; Messenger RNA; Methodology; Methods; Microscopy; Molecular; Molecular Probes; Monitor; Neurobiology; Neurons; Neurosciences; Organism; Proteins; Recombinants; Reporting; Resolution; Site; Sleep; Solutions; Structure; Synapses; Synaptic plasticity; System; Techniques; Technology; Testing; Time; Tissue Extracts; Vertebrates; Zebrafish; addiction; base; conditioning; data structure; fluorescence imaging; gamma-Aminobutyric Acid; gephyrin; imaging probe; in vivo; innovation; light microscopy; novel; postsynaptic; preference; presynaptic; public health relevance; relating to nervous system; research study; software development; sound; submicron; tool; two-photon

DESCRIPTION (provided by applicant): Here we propose to develop an experimental paradigm to allow dynamic monitoring of the strength and location of every glutamatergic and GABA/Glycinergic synapse within the brain of a living organism. In combination with behavioral manipulation of the organism this paradigm will allow for study of how the brain encodes information in synaptic structure. This paradigm will involve combining three technologies: 1. Recombinant probes with which postsynaptic excitatory and inhibitory sites can be labeled in vivo, allowing the location and strength of synaptic connections to be monitored in parallel. 2. 2P-SPIM microscopy, which can image large volumes very quickly, without bleaching and, potentially, with isotropic resolution. 3. Software to calculate and store the location and strengt of each synapse in such a manner that it can be easily manipulated and analyzed. Experiments will be performed in zebrafish, as they have semi-transparent brains that are relatively small, yet they are capable of relatively complex behaviors. Experiments will be used both to establish the viability of the paradigm and to answer fundamental questions about how synapses are modulated during sleep, as well as how they are changed in learning paradigms such as sound habituation and place preference/aversion conditioning.

描述（由申请人提供）：在这里，我们建议开发一个实验范式，以动态监测活生物体大脑内每个谷氨酸能和GABA和GABA/GLABA/GLYCIN能突触的强度和位置。结合对生物的行为操纵，这种范式将允许研究大脑如何编码突触结构中的信息。该范式将涉及组合三种技术：1。可以在体内标记突触后兴奋性和抑制性位点的重组探针，从而可以并行监测突触连接的位置和强度。 2。2p-Spim显微镜可以很快地对大量摄像，而无需漂白，并且可能具有各向同性分辨率。 3.软件以一种可以轻松操纵和分析的方式来计算和存储每个突触的位置和strengt。实验将在斑马鱼中进行，因为它们的半透明大脑相对较小，但 他们能够采取相对复杂的行为。实验将既用于建立范式的生存能力，又要回答有关在睡眠期间如何调节突触的基本问题，以及如何在学习范式（例如健全习惯和放置偏好/厌恶条件）中改变它们。