Developing Novel Trans-Synaptic Viral Vectors for Orthogonal or Rapid Circuit Tracing

项目摘要 要确定复杂神经行为的解剖学基础，关键是有能力 在同一动物身上同时追踪多个回路。那是因为复杂的动物 行为或神经计算应该通过一个以上的交互作用来理解 巡回合作、对抗或其他。此外，有必要迅速捕获 在发育和学习过程中动态变化的大脑中的连接信息。 基因工程G-缺失狂犬病是一种目前最先进的方法，可以逆行追踪突触前 定义的细胞类型的输入神经元。然而，追踪一个以上的神经仍然是不可行的。 同时巡回。此外，目前使用AAV助手和狂犬病的方法需要 需要几个星期才能追踪到。在这项拟议的研究中，我们将通过以下方式克服这些缺点 开发两种新的跨突触病毒示踪系统：剑：仙台与正交 狂犬病双链追踪(目标1)和快速Trio/cTRIO：细胞类型特异性追踪关系 在投入和产出之间(目标2)。这项研究具有重要意义，因为这些新方法将 允许在多个电路和多个电路中进行更全面的神经连接分析 不同的背景，例如发育中的大脑，在那里出现不同的突触网络和神经 可塑性，例如在许多模式物种中学习。这项拟议的研究具有创新性， 因为我们正在开发和验证技术创新的解决方案，剑和快速 Trio/cTRIO，以克服当前最先进的跟踪方法的局限性。这些 病毒遗传工具将对神经科学领域产生积极而广泛的影响 加深对复杂行为的神经回路组织的认识，有助于 确定治疗脑部疾病的特定电路治疗靶点。

Project Summary To determine the anatomical basis of complex neural behavior, it is critical to have the ability to trace more than one circuit simultaneously in the same animal. That’s because complex animal behaviors or neural computation should be understood through the interaction of more than one circuit – cooperative, antagonistic, or else. In addition, it is necessary to rapidly capture the connectivity information in the dynamically changing brains during development and learning. Engineered G-deleted rabies is a current state-of-art method to retrogradely trace the presynaptic input neurons of a defined cell type. However, it remains unfeasible to trace more than one neural circuit simultaneously. In addition, the current approach using AAV helpers and rabies requires several weeks for tracing. In this proposed research, we will overcome these disadvantages by developing two novel trans-synaptic viral tracer systems: SWORD: Sendai with Orthogonal Rabies Duplex Tracing (Aim 1) and a rapid TRIO/cTRIO: cell-type specific tracing the relationship between input and output (Aim 2). This research is significant because these new methods will allow more comprehensive analysis of neural connectivity in more than one circuit and in more diverse context such as the developing brain where distinct synaptic networks emerge and neural plasticity such as learning across many model species. The proposed research is innovative, because we are developing and validating technically innovative solutions, SWORD and rapid TRIO/cTRIO, to overcome the limitations of the current state-of-the art tracing method. These viral-genetic tools will have a positive and broad impact on the neuroscience field as it will enhance our understanding of neural circuit organization for the complex behaviors and help to identify the circuit-specific therapeutic targets to cure brain disorders.