Optogenetic mapping of synaptic activity and control of intracellular signaling

突触活动的光遗传学图谱和细胞内信号传导的控制

• 批准号: 8827155
• 负责人: David Kleinfeld
• 金额: $ 38.75万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-30 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8827155
• 关键词: Adaptive Behaviors; Adenylate Cyclase; Adrenergic Agents; Adrenergic Receptor; Alzheimer' s Disease; Amino Acid Motifs; Amygdaloid structure; Animals; Behavior; Behavioral; Binding; Brain; Brain Mapping; Brain region; Cells; Code; Communication; Competitive Binding; Complement; Corpus striatum structure; Coupled; Development; Dopamine; Dorsal; Drug Addiction; Fluorescence; Fright; G-Protein-Coupled Receptors; GTP-Binding Proteins; Generations; Glutamates; Goals; Heterodimerization; Intracellular Second Messenger; Lateral; Lead; Learning; Light; Lighting; Long-Term Depression; Long-Term Potentiation; Maps; Masks; Mediating; Membrane; Memory; Methods; Molecular; Nerve Degeneration; Nervous system structure; Neurodegenerative Disorders; Neurologic; Neuromodulator; Neurons; Neuropeptides; Neurosciences; Neurotransmitters; Optics; Pathway interactions; Peptide Hydrolases; Peptides; Performance; Plastics; Population; Postsynaptic Membrane; Presynaptic Terminals; Probability; Protein Fragment; Proteins; Protocols documentation; Receptor Activation; Receptor Inhibition; Reporter; Reporting; Research Personnel; Resolution; Rodent; Role; Second Messenger Systems; Sensory; Signal Transduction; Site; Specificity; Synapses; Synaptic Cleft; Synaptic plasticity; System; Techniques; Testing; Time; Traumatic Brain Injury; Venus; Vesicle; addiction; adrenergic; base; conditioned fear; extracellular; in vivo; neuronal excitability; neurotransmitter release; optogenetics; postsynaptic; promoter; protein activation; public health relevance; receptor coupling; reconstitution; second messenger; tool

 DESCRIPTION (provided by applicant): This proposal aims to develop new molecular techniques to map activities of neurons, manipulate the strength of communication between neurons and disrupt intracellular signaling. These 'optogenetic' approaches will be used to further our understandings of brain function on behavior and have important implications in our understandings of neurological conditions and neurodegenerative diseases. The first goal is to develop a technique where the researchers can use optical approach to identify synaptic connections that were active during the performance of a behavior task. This reporter system can be turned on with light, which defines the window of activity reporting, and fluorescence signal can be detected if there is significant activity between two defined cell groups. Many existing approaches can only be used to map excitatory connections, whereas the proposed approach can be used to identify activities between synapses utilizing any neurotransmitters. The approach will utilize a split fluorescent protein approach where its complementation and the generation of fluorescent signal is activity dependent. This approach will test whether a defined synaptic connection is involved in the performance of a behavior. The second goal is to develop a technique where the researchers can use light to modulate the strength of synaptic communication between neurons. Increasing synaptic strength is believed to underlie memory and learning, and its disruption has been implicated in drug addiction and many neurological conditions. Having the ability to modulate the synaptic strength experimentally can be used to interrogate how changes in synaptic strength alter learning and memory, leading to the observed adaptive behavior in the animals in both normal and pathological conditions. Many small protein fragments can alter synaptic strengths between neurons. A light-responsive protein can be used to functionally mask these protein fragments in the dark and light can be used to functionally release these protein fragments. This will permit rapid experimental control of synaptic strength and their functional effects can be studied in the behaving animals. This tool can be used to understand how alteration in synaptic strength changes during learning and adaption. The third goal of the project is to develop a technique where G-protein coupled receptor mediated second messenger pathway is inhibited by light. G-protein coupled receptors mediate the effects of neuromodulator and neuropeptides in the nervous system and they have great importance in modulating and/or mediating behaviors. Using a similar approach as described above, competitive binding peptides that disrupt G-protein coupled receptor-G protein interactions or peptides that directly inhibit the effectors of G- protein pathways can be masked and unmasked with light-responsive protein and light illumination. With this approach, light will turn off G protein activation or effectors of G-protein pathway rapidly to interrogate the behavioral effects of neuromodulators or neuropeptides in specific cells with defined temporal resolution.

 描述（由申请人提供）：该提案旨在开发新的分子技术来绘制神经元的活动图，操纵神经元之间的通信强度并破坏细胞内信号传导。这些“光遗传学”方法将用于进一步了解大脑行为功能，并对我们理解神经系统疾病和神经退行性疾病具有重要意义。 第一个目标是开发一种技术，研究人员可以使用光学方法来识别在执行行为任务期间活跃的突触连接。该报告系统可以用光打开，这定义了活动报告的窗口，如果两个定义的细胞组之间存在显着的活动，则可以检测到荧光信号。许多现有方法只能用于绘制兴奋性连接，而所提出的方法可用于利用任何神经递质来识别突触之间的活动。该方法将利用分裂荧光蛋白方法，其互补和荧光信号的产生是活性依赖性的。这种方法将测试定义的突触连接是否参与行为的表现。 第二个目标是开发一种技术，研究人员可以利用光来调节神经元之间突触通讯的强度。突触强度的增强被认为是记忆和学习的基础，其破坏与毒瘾和许多神经系统疾病有关。通过实验调节突触强度的能力可用于探究突触强度的变化如何改变学习和记忆，从而导致在正常和病理条件下观察到动物的适应性行为。许多小蛋白质片段可以改变神经元之间的突触强度。光响应蛋白可用于在黑暗中功能性地掩盖这些蛋白片段，而光可用于功能性地释放这些蛋白片段。这将允许对突触强度进行快速实验控制，并且可以在行为动物中研究它们的功能效果。这个工具 可用于了解学习和适应过程中突触强度的变化如何变化。 该项目的第三个目标是开发一种光抑制 G 蛋白偶联受体介导的第二信使途径的技术。 G蛋白偶联受体介导神经系统中神经调节剂和神经肽的作用，并且它们在调节和/或介导行为中具有非常重要的作用。使用如上所述的类似方法，可以用光响应蛋白和光照射来掩蔽和揭露破坏G蛋白偶联受体-G蛋白相互作用的竞争性结合肽或直接抑制G蛋白途径效应子的肽。通过这种方法，光将快速关闭 G 蛋白激活或 G 蛋白通路效应器，以限定时间分辨率询问特定细胞中神经调节剂或神经肽的行为效应。