Viral Brainbow: tracing brain circuits with connection order specificity

Viral Brainbow：追踪具有连接顺序特异性的大脑回路

• 批准号: 7817417
• 负责人: John Patrick Card
• 金额: $ 50万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7817417
• 关键词: Address; Animals; Area; Behavior; Binding; Brain; Calcium; Cells; Color; Communities; DNA Viruses; Data; Generations; Genetic Recombination; Genome; Herpesviridae; Image; In Vitro; Methods; Microfluidic Microchips; Modification; Nervous System Physiology; Neurons; Neurosciences; Population; Property; Proteins; Rattus; Reporter; Reporting; Research Project Grants; Site; Specificity; Speed; Suid Herpesvirus 1; System; Technology; Testing; Tracer; Transgenic Mice; Transgenic Organisms; Viral; Virus; Virus Diseases; Work; base; calcium indicator; design; fundamental research; in vivo; insight; neural circuit; novel; presynaptic; prototype; recombinase; relating to nervous system; research study; sensor; tool

DESCRIPTION (provided by applicant): This application addresses broad Challenge Area (06) Enabling Technologies, 06-NS-106: Validating new methods to study brain connectivity. We propose to test a new method that provides substantial improvement over previous Cre-conditional viral tracers. The technology combines the Brainbow multicolor cell marking technology with the retrograde, circuit tracing properties of pseudorabies virus (PRV), a neuroinvasive alpha herpesvirus. We have constructed a prototype PRV Brainbow virus called PRV263 that we propose will enable simultaneous identification of distinct chains of neurons projecting to a phenotypically defined population of neurons, and promises to provide predictive data on the strength of different connections among those neurons. Importantly, these PRV Brainbow tracers will have distinct advantages over present tracers. Our concept takes advantage of conditional, site-specific recombination of the genome of a DNA virus to produce multiple reporters so that neurons upstream (presynaptic) of a Cre recombinase (Cre) expressing neuron will be a different color from the Cre-expressing neuron. This novel concept will be expanded to produce second generation prototypes of PRV Brainbow tracers that do not rely on Cre-transgenic mice and can be used in the many mammalian species susceptible to PRV infection. A third generation prototype will be constructed that not only marks circuits, but also reports on neuronal activity. In this latter concept, the PRV Brainbow virus also will include a genetically encoded calcium indicator that fluoresces when calcium is bound. As viral tracing of neural circuitry has become an essential tool in the neuroscience community, our new tracers will be immediately applicable for many ongoing fundamental research projects in neuroscience in a variety of animals. These new tools have promise to reveal detailed functional insights into neural circuit organization that have not been possible to achieve in the past. Viral tracing of neural circuitry has become an essential tool in the neuroscience community. The new, robust viral tracers that will result from our work have promise to reveal detailed functional insights into trans-neuronal spread of herpesviruses, as well as neural circuit organization that have not been possible to achieve in the past. These neural tracers would be powerful tools to elucidate brain micro-circuitry, providing a better understanding of nervous system functions.

描述（由申请人提供）：此申请解决广泛的挑战领域（06）启用技术，06-NS-106：验证研究大脑连通性的新方法。 我们建议测试一种新方法，该方法比以前的Cre-Conditional病毒示踪剂可提供大量改进。 该技术将Brainbow多色细胞标记技术与逆行的逆行跟踪特性（PSEUDORABIES病毒（PRV）（一种神经性αα疱疹病毒）相结合。 我们已经构建了一种称为PRV263的原型PRV脑弓病毒，我们建议将同时识别投射到表型定义的神经元种群的神经元的不同链，并有望提供有关这些神经元之间不同连接强度的预测数据。 重要的是，这些PRV Brainbow示踪剂将在当前的示踪剂上具有明显的优势。 我们的概念利用了DNA病毒基因组的条件，特定于位点的重组来产生多个报告基因，以使表达神经元的CRE重组酶（CRE）上游（突触前）的神经元与表达CRE的神经元具有不同的颜色。 这个新颖的概念将扩展，以生产不依赖Cre-Transgenic小鼠的PRV Brainbow示踪剂的第二代原型，并且可用于许多容易受到PRV感染的哺乳动物物种。 将构建第三代原型，不仅标记电路，而且还报告了神经元活动。 在后一种概念中，PRV脑弓病毒还将包括一个遗传编码的钙指标，该指标在钙结合时会产生荧光。 随着神经电路的病毒追踪已成为神经科学界的重要工具，我们的新示踪剂将立即适用于许多正在进行的动物神经科学中的基本研究项目。 这些新工具有望揭示对过去无法实现的神经电路组织的详细功能见解。 神经电路的病毒追踪已成为神经科学界的重要工具。 我们工作将导致的新的，健壮的病毒示踪剂有望揭示疱疹病毒跨性神经元扩散的详细功能见解，以及过去无法实现的神经电路组织。 这些神经示踪剂将是阐明脑微路的强大工具，可以更好地理解神经系统功能。