A genetic strategy to record cell-cell interactions

记录细胞间相互作用的遗传策略

• 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
• 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

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）：理解大脑回路中发生的计算需要识别这些回路中的神经元如何相互连接。近年来，一些新的方法（最值得注意的是，连续电子显微镜，复制缺陷型狂犬病病毒，和GRASP）已被设计来识别大脑回路的接线图。为了克服目前可用的策略的一些限制，我们建议生成一个新的遗传编码系统，通过跨突触控制转录来跟踪大脑回路，这将为研究回路连接和功能之间的关系开辟新的机会。我们提出的系统是基于Notch受体的分子逻辑。在该系统中，表达人工配体的神经元（“发送者”神经元）激活其突触伴侣（“接收者”神经元）上的遗传修饰的Notch受体。在突触位点中的配体-受体相互作用后，工程化受体在其跨膜结构域中被切割并释放调节突触伴侣中转录的蛋白质片段。我们最初的体外实验已经证实了这种策略的可行性，我们建议将这种设计应用于识别转基因动物神经元回路的接线图，包括小鼠和果蝇。