Dynamic mapping of the complete synaptome using recombinant probes

使用重组探针动态绘制完整突触组

基本信息

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
项目类别：
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

项目摘要

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，我们建议开发一种实验范例，以允许动态监测活体大脑内每个谷氨酸能和 GABA/甘氨酸能突触的强度和位置。与生物体的行为操纵相结合，该范式将允许研究大脑如何在突触结构中编码信息。这一范例将涉及三种技术的结合： 1. 重组探针，可以在体内标记突触后兴奋性和抑制性位点，从而可以并行监测突触连接的位置和强度。 2. 2P-SPIM 显微镜，可以非常快速地对大体积图像进行成像，无需漂白，并且可能具有各向同性分辨率。 3. 以易于操作和分析的方式计算和存储每个突触的位置和强度的软件。实验将在斑马鱼中进行，因为它们具有相对较小的半透明大脑，但他们能够做出相对复杂的行为。实验将用于建立该范式的可行性，并回答有关突触在睡眠期间如何调节的基本问题，以及突触如何在学习范式中发生变化，例如声音习惯化和位置偏好/厌恶调节。