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

项目摘要 为了确定复杂神经行为的解剖学基础，关键是要有能力 在同一动物身上同时追踪一个以上的回路。这是因为复杂的动物 行为或神经计算应该通过一个以上的相互作用来理解。 电路-合作，对抗，或其他。此外，还必须迅速捕捉 在发育和学习过程中，大脑中动态变化的连接信息。 工程G-缺失狂犬病是目前最先进的方法，可以追溯突触前 定义的细胞类型的输入神经元。然而，追踪一个以上的神经元仍然是不可行的。 电路同时此外，目前使用AAV助手和狂犬病的方法需要 几个星期的追踪。在这项研究中，我们将克服这些缺点， 开发两种新型跨突触病毒示踪系统：SWORD：Sendai与正交 狂犬病双链体示踪（Aim 1）与快速TRIO/cTRIO：细胞类型特异性示踪的关系 输入和输出之间的关系（目标2）。这项研究意义重大，因为这些新方法将 允许更全面地分析多个回路和多个回路中的神经连接 不同的背景，如发育中的大脑，其中出现了不同的突触网络和神经网络， 可塑性，例如在许多模型物种中学习。这项研究具有创新性， 因为我们正在开发和验证技术创新的解决方案， TRIO/cTRIO，以克服目前最先进的追踪方法的局限性。这些 病毒遗传工具将对神经科学领域产生积极和广泛的影响， 增强我们对复杂行为的神经回路组织的理解，并有助于 确定特定回路的治疗靶点，以治愈脑部疾病。

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.