A genetic strategy to record cell-cell interactions

记录细胞间相互作用的遗传策略

基本信息

批准号：
8681568
负责人：
CARLOS LOIS
金额：
$ 20.73万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8681568
关键词：
Afferent Neurons Animals Antibodies Antigens Autistic Disorder Axon Brain Calcium Cell Communication Cell Nucleus Cells Cleaved cell Dendrites Development Drosophila genus Electron Microscopy Engineering Genetic Genetic Transcription Goals In Vitro Invertebrates Label Lead Lentivirus Vector Ligand Binding Domain Ligands Lobe Logic Measurement Methods Modification Molecular Mus Neurodevelopmental Disorder Neurons Notch Signaling Pathway Pathway interactions Physiological Protein Fragment Rabies virus Reporter Research Research Personnel Risk Schizophrenia Site Speed Synapses System Transgenes Transgenic Animals Transgenic Mice Transgenic Organisms Transmembrane Domain Vertebrates base cell type design fly genetic manipulation in vivo neuronal cell body notch protein novel strategies optogenetics postsynaptic promoter public health relevance receptor research study sensor

项目摘要

DESCRIPTION (provided by applicant): Understanding the computations that take place in brain circuits requires identifying how neurons in those circuits are connected to each other. In recent years several new approaches (most notably, serial electron microscopy, replication-deficient rabies viruses, and GRASP) have been designed to identify the wiring diagrams of brain circuits. To overcome some of the limitations of the currently available strategies we propose to generate a new genetically-encoded system to trace brain circuits by transsynaptic control of transcription that will open new opportunities for investigating the relationship betwee circuit connectivity and function. The system that we propose is based on the molecular logic of the Notch receptor. In this system, neurons expressing an artificial ligand ("sender" neurons) activate a genetically-modified Notch receptor on their synaptic partners ("receiver" neurons). Upon ligand-receptor interaction in synaptic sites, the engineered receptor is cleaved in its transmembrane domain and releases a protein fragment that regulates transcription in the synaptic partners. Our initial experiments in vitro have confirmed the feasibility of this strategy and we propose to apply this design towards identifying wiring diagrams of neuronal circuits in transgenic animals, both in mice and drosophila.

描述（由申请人提供）：了解大脑电路中发生的计算需要确定这些电路中的神经元如何相互连接。近年来，已经设计了几种新方法（最值得注意的是，串行电子显微镜，复制缺陷的狂犬病病毒和掌握）旨在识别脑电路的接线图。为了克服当前可用策略的某些局限性，我们建议通过转录的转录控制来生成新的遗传编码系统来追踪脑电路，这将为研究之间的关系开放新的机会，以研究之间的关系。我们提出的系统基于Notch受体的分子逻辑。在该系统中，表达人工配体（“发送者”神经元）的神经元在其突触伴侣（“接收器”神经元）上激活了遗传改性的Notch受体。在突触部位的配体 - 受体相互作用时，工程受体在其跨膜结构域中裂解，并释放出调节突触伙伴中转录的蛋白质片段。我们在体外的最初实验证实了该策略的可行性，我们建议将这种设计应用于小鼠和果蝇中转基因动物中神经元电路的接线图。