Novel Genetically Encoded Indicators for Interrogating Neuron-Astrocyte Communication Across Timescales

用于询问神经元-星形胶质细胞跨时间尺度通讯的新型基因编码指标

• 批准号: 10294806
• 负责人: Lin Tian
• 金额: $ 42万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-15 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10294806
• 关键词: Acetylcholine; Address; Affect; Astrocytes; Axon; Behavior; Behavioral; Biology; Biosensor; Brain; Calcium; Cell Adhesion Molecules; Color; Communication; Complex; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Development; Dopamine; Engineering; Equilibrium; Excitatory Synapse; Fluorescence; G-Protein-Coupled Receptors; Genomics; Glutamates; Image; Individual; Infiltration; Inhibitory Synapse; Investigation; Knowledge; Label; Lead; Measures; Mind; Modeling; Modernization; Molecular; Mus; Neuromodulator; Neurons; Neurotransmitters; Pathology; Pathway interactions; Pattern; Pharmaceutical Preparations; Phosphotransferases; Process; Property; Protein Engineering; Proteins; Resolution; Resources; Role; Serotonin; Shapes; Signal Pathway; Signal Transduction; Slice; Stimulus; Structure; Supporting Cell; Synapses; Synaptic Transmission; Techniques; Testing; awake; base; calcium indicator; cell type; cognitive process; design; experience; experimental study; extracellular; gamma-Aminobutyric Acid; genetic manipulation; imaging system; improved; in vivo; minimally invasive; multidisciplinary; multiplexed imaging; neural circuit; neuronal circuitry; neuroregulation; novel; novel strategies; optogenetics; postsynaptic; reconstitution; response; screening; sensor; spatiotemporal; synaptic function; synaptogenesis; tool; two photon microscopy; two-photon

Project Summary: Project 4 - Novel Genetically Encoded Indicators for Interrogating Neuron-Astrocyte Communication Across Timescales Astrocytes, the most abundant cell type in the brain, have long thought to be primarily passive support cells. Considerable evidence from the labs has shown that astrocyte-synapse displays a dynamic and bi-directional relationship, with local synaptic transmission and neuromodulation being capable of shaping astrocytic activity and PAP structural plasticity, and astrocyte shaping synapse formation and modulating plasticity and signaling via secreted factors and adhesion molecules. These critical advances in understanding astrocyte biology in vivo are primarily due to recent applications of modern techniques initially designed for studying neurons to direct manipulation and interrogation of astrocytes. Though the concept of astrocytes as integral and modulatory components of neural circuit is emerging, a mechanistic understanding of causative and correlative roles of astrocytes in operating neural circuit and contribution to the complex behaviors is still lacking, which necessities and drives the development of improved tools. Thus, a large-scale protein engineering effort to develop an improved tool to address unsolved questions to achieve a mechanistic understanding of causal and correlative roles of astrocyte in neuronal circuit function and contributions to behavior is being proposed. Provided items include: 1. a set of optimized red-shifted glutamate, GABA, DA, and NE sensors, 2. a set of green and red-shifted synaptic glutamate/GABA sensors to probe neuron-astrocyte connectivity and extracellular NT transients at tripartite synapses, and 3. interrogate cross-talk between PKA and calcium in astrocytes and optimized green and red-shifted kinases sensors for in vivo applications. These new sensors will be applied to study 1) how experience-dependent changes that drive complex patterns of neurotransmitter or neuromodulatory signaling lead to the changes in astrocytic activity and 2) how astrocytes modulate synaptic activity via structural plasticity across various temporal scales. The contribution is significant because these improved tools will permit new hypotheses being tested in astrocyte biology. This toolset will provide needed tools to facilitate experiments proposed here and provide a rich resource to the field to bring full swing the investigation of astrocyte-neuron interaction underlying complex behavioral and cognitive processes that are inaccessible via currently existing approaches.

项目摘要：项目 4 - 用于询问神经元星形胶质细胞的新型基因编码指标 跨时间尺度的沟通 星形胶质细胞是大脑中最丰富的细胞类型，长期以来一直被认为主要是被动支持细胞。 来自实验室的大量证据表明，星形胶质细胞突触表现出动态和双向的 关系，局部突触传递和神经调节能够塑造星形胶质细胞的活动 和 PAP 结构可塑性，星形胶质细胞塑造突触形成并调节可塑性和信号传导 通过分泌因子和粘附分子。这些在理解星形胶质细胞生物学方面的关键进展 体内主要归功于现代技术的最新应用，这些技术最初是为研究神经元而设计的 星形胶质细胞的直接操作和询问。尽管星形胶质细胞的概念是完整且 神经回路的调节成分正在出现，对因果关系和相关关系的机械理解 星形胶质细胞在操作神经回路中的作用以及对复杂行为的贡献仍然缺乏，这 必要性并推动改进工具的开发。因此，大规模的蛋白质工程努力 开发一种改进的工具来解决未解决的问题，以实现对因果关系的机械理解 星形胶质细胞在神经元回路功能和对行为的贡献中的相关作用正在被提出。 提供的物品包括： 1.一组优化的红移谷氨酸、GABA、DA和NE传感器， 2. 一组绿移和红移突触谷氨酸/GABA传感器，用于探测神经元-星形胶质细胞的连接 和三方突触处的细胞外 NT 瞬变，以及 3.询问星形胶质细胞中PKA和钙之间的串扰并优化绿移和红移 用于体内应用的激酶传感器。 这些新传感器将用于研究 1) 经验依赖的变化如何驱动复杂的模式 神经递质或神经调节信号传导导致星形胶质细胞活动的变化以及 2) 如何 星形胶质细胞通过不同时间尺度的结构可塑性调节突触活动。贡献是 意义重大，因为这些改进的工具将允许在星形胶质细胞生物学中测试新的假设。这 工具集将提供所需的工具来促进此处提出的实验，并为该领域提供丰富的资源 全面展开复杂行为和认知背后的星形胶质细胞-神经元相互作用的研究 通过当前现有方法无法访问的过程。