Fluorescent Protein Sensors of Neural Activity

神经活动的荧光蛋白传感器

• 批准号: 6857791
• 负责人: Ehud Isacoff
• 金额: $ 34.13万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-03-15 至 2009-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6857791
• 关键词: NMDA receptors; Xenopus; bioimaging /biomedical imaging; biological signal transduction; calcium ion; confocal scanning microscopy; electrical potential; fluorescent dye /probe; gene expression; genetically modified animals; hippocampus; laboratory mouse; laboratory rat; membrane potentials; molecular assembly /self assembly; neural information processing; neurogenetics; neuroimaging; neuromuscular junction; psychomotor function; reporter genes; second messengers; synapses; tissue /cell culture

DESCRIPTION (provided by applicant): The pattern of activity in the circuits of the brain and their experience-dependent changes underlie the processing of sensory information, perception, and memory. Much has been learned about the anatomical wiring of brain circuits and about the receptive field properties of individual neurons in the intact brain, but considerable mystery remains about how the properties of individual neurons emerge from their connectivity and how their activity is assembled into coherent percepts. Part of the problem has been that high precision recording is usually obtained from only one or a few neurons at a time, when salient events are actually processed by large assemblies of neurons. The method with the best resolution in time and space currently for population recording is the optical recording of neural activity with voltage-sensitive chemical dyes. However, these optical reports are blurred by their lack of targeting to specific cell types or subcellular compartments. Glial signals are confounded with ones from neurons, distinct types of neurons contribute indistinguishably to the signal, and the neuronal component of the signal is dominated by the dendrites, which represent the major membrane area. What has been missing is a method of targeting these optical reporters. The goal of this grant is to create genetically-encoded optical reporters of neural activity to fill this gap. The advantage of this approach is that the reporters can be specifically expressed in desired cell types through the use of cell-type-specific promoters, and they can be targeted to specific subcellular compartments. Our emphasis is to develop reporters that reveal action potential firing, synaptic activity, second messenger signaling, and changes in the molecular assembly of presynaptic active zones and postsynaptic densities. These reporters will be expressed in cultured cells to optimize their properties and then delivered transgenically and via viruses to model systems (the Drosophila larval neuro-muscular junction, and the mouse and rat barrel cortex) in which they will be tested in vitro and in vivo.

描述（由申请人提供）：大脑电路中的活动模式及其与经验有关的变化是感官信息，感知和记忆的处理。关于脑电路的解剖接线以及完整大脑中单个神经元的接受场特性已经了解了很多，但是关于单个神经元的特性如何从它们的连通性中出现以及它们的活性如何组装到相干感知中。问题的一部分是，当大型神经元组合实际处理显着事件时，通常只从一个或几个神经元中获得高精度记录。目前记录的时间和空间分辨率最佳的方法是对电压敏感化学染料的神经活动的光学记录。但是，这些光学报告由于缺乏针对特定细胞类型或亚细胞室的靶向而变得模糊。神经胶质信号与神经元的神经元混淆，不同类型的神经元对信号的贡献是没有区别的，并且信号的神经元成分由树突支配，这代表了主要的膜面积。缺少的是针对这些光学记者的方法。这项赠款的目的是创建神经活动的遗传编码的光学记者来填补这一空白。这种方法的优点是，可以通过使用细胞型特异性启动子在所需的细胞类型中特异性表达记者，并且可以针对特定的亚细胞隔室。我们的重点是开发记者，这些记者揭示了动作潜在的发射，突触活动，第二信号信号传导以及突触前活性区和突触后密度的分子组装的变化。这些记者将在培养的细胞中表达，以优化其特性，然后通过病毒和病毒传递到模型系统（果蝇幼虫神经肌肉连接点，小鼠和大鼠桶皮层），在该系统中将在体外和体内测试它们。