Sparse, Strong and Large Area Targeting of Genetically Encoded Indicators

遗传编码指标的稀疏、强和大面积目标

• 批准号: 9146401
• 负责人: SRDJAN D ANTIC
• 金额: $ 53.88万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-18 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9146401
• 关键词: Adverse effects; Algorithms; Animals; Antibiotics; Area; BRAIN initiative; Blindness; Brain; Calcium; Cells; Cognitive; Craniotomy; Data; Data Analyses; Development; Disease; Dyes; Electrodes; Emotional; Event; Funding; Genetic; Genetic Recombination; Glass; Goals; Golgi Apparatus; Health; Image; Imaging Device; Imaging problem; Individual; Label; Life; Maps; Measurement; Mediating; Mental disorders; Metals; Methodology; Methods; Microscopy; Molecular; Monitor; Motor; Mus; Nerve Tissue; Neuraxis; Neurons; Neurosciences; Noise; Optics; Pattern; Population; Population Heterogeneity; Probability; Procedures; Process; Protocols documentation; Pyramidal Cells; Reporting; Resolution; Scanning; Signal Transduction; Slice; Somatosensory Cortex; Staining method; Stains; Symptoms; Synaptic Potentials; System; Techniques; Technology; Time; Tissues; Transgenic Organisms; Translating; Trimethoprim; Work; base; cellular targeting; data acquisition; design; in vivo; information processing; interest; invention; neuronal circuitry; neurophysiology; neurotechnology; novel; preempt; protein expression; recombinase; reconstruction; response; segregation; sensory stimulus; small molecule; spatiotemporal; two-photon; voltage

 DESCRIPTION (provided by applicant): Electrical (voltage) signal is the primary substrate of information processing in the brain. Detecting and recording voltage changes from neurons in living animals remains the ultimate goal of experimental neuroscience to the present day. Standard glass and metal electrodes are hugely invasive and their use suffers from poor spatial resolution, limited coverage, and blindness to cellular identity. The popular calcium-sensitive indicators generate signals contaminated with changes in intracellular calcium that are unrelated to neuronal electrical signals, and only indirectly report the electrical signals with distorted tiing and highly distorted waveform. A conceptually ideal principle to achieve monitoring of neuronal electrical activity is provided by optical voltage imaging using genetically-encoded voltage indicators (GEVIs). However, even the best performing GEVIs currently available are suitable only for in vivo monitoring of compound synaptic potentials (the summated voltage signal from unidentified number of neurons), but fail to resolve simultaneously signals from many individual cells in intact nervous tissue. While better performing GEVIs with higher sensitivity are expected to emerge from the on-going BRAIN Initiative-funded activities, this alone does not resolve the single-cell resolution voltage imaging problem: signals from individual cells are not spatially segregated. The solution to this issue requires other technologies including refined genetic targeting and data analysis methods. We plan to develop protocols for sparse GEVI targeting of central nervous system neurons. Sparse cellular targeting will allow imaging of neuronal activity with little spatial overlap. We also plan to develop data analysis routines for isolating single cel responses from data obtained in sparsely labelled tissue and, building on this, in densely targeted tissue. In summary, we propose to develop a novel genetically-directed voltage imaging tool that is qualitatively different than those currently available, along with data analyss methods to facilitate the use of electrical signals from large number of neurons embedded in functioning neuronal networks. The novel GEVI labelling protocols and data acquisition and analysis algorithms developed here should be immediately useful and impactful for studying brain function in health and disease.

 描述（由适用提供）：电气（电压）信号是大脑信息处理的主要基板。在当今，从活跃动物中的神经元中检测和记录电压变化仍然是实验性神经科学的最终目标。标准的玻璃和金属电极具有极大的侵入性，其使用遭受了差的空间分辨率，覆盖范围有限和对细胞身份的失明。流行的钙敏感指标产生的信号被细胞内钙的变化污染，这些变化与神经元电信号无关，并且仅间接地报告了带有扭曲的层次和高度扭曲的波形的电信号。通过遗传编码的电压指示剂（GEVIS），通过光电压成像（GEVIS）提供了一个概念上的理想原理，以实现神经元电活动的监测。但是，即使是最佳性能GEVIS，目前都可以使用，仅适用于体内监测复合突触电位（来自未识别的神经元的总和电压信号），但无法同时解决来自完整神经组织中许多单个细胞的信号。虽然预计较高灵敏度的GEVI可以从持续的大脑启动资助的活动中出现，但仅此功能并不能解决单细胞分辨率电压成像问题：来自单个细胞的信号并未在空间上隔离。解决此问题的解决方案需要其他技术，包括精致的遗传靶向和数据分析方法。我们计划开发用于稀疏GEVI靶向中枢神经系统神经元神经元的方案。稀疏的细胞靶向将允许在几乎没有空间重叠的情况下对神经元活性进行成像。我们还计划开发数据分析程序，以从稀疏标记的组织中获得的数据隔离单个CEL响应，并在其基础上建立在固定的靶向组织中。总而言之，我们建议开发一种新型的一般指导的电压成像工具，该工具与当前可用的电压截然不同，以及数据分析方法，以促进来自嵌入神经元网络中大量神经元的电信号的使用。在此开发的新型GEVI标记方案以及数据获取和分析算法应立即有用且有影响力，可用于研究健康和疾病中的大脑功能。