Mapping Neural Circuits Using Pseudorabie Virus Vectors

使用伪狂犬病病毒载体绘制神经回路

• 批准号: 6857740
• 负责人: JEFFREY M FRIEDMAN
• 金额: $ 39.88万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-02-15 至 2009-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6857740
• 关键词: brain mapping; gene expression; genetically modified animals; hypothalamus; laboratory mouse; method development; neuroanatomy; neurons; polymerase chain reaction; recombinant virus; reporter genes; southern blotting; suid alphaherpesvirus 1; transfection /expression vector

DESCRIPTION (provided by applicant): This proposal outlines the development of novel methods for the mapping of complex neural pathways and the specification of the neurons that compose these pathways. Our groups have recently made a recombinant Pseudorabies virus (PRV) that is conditional on cre-recombinase for viability and expression of a GFP reporter gene. This virus has been used successfully to map inputs to specific neuronal populations in defined brain regions. For simplicity we will refer to this and related methods as PAM, Pseudorabies virus assisted mapping. In this application, we propose to further develop PAM so as to allow: 1) Simultaneous Retrograde Tracing from Different Neurons in the Same Brain Region, 2) Simultaneous Retrograde Tracing from Similar Neurons in Different Brain Regions, 3) Anterograde tracing 4) Identification of Neurons Projecting to More than One Site 5) Specification of the neurons that are identified in these mapping studies. To establish the validity of the approach, these studies will be performed in analyses of hypothalamic circuits that regulate feeding. Once the methods are validated, the viruses and animals will be made freely available to other investigators interested in analogous studies of other neuronal pathways.

描述（由申请人提供）：该提案概述了复杂神经通路映射和组成这些通路的神经元规范的新方法的开发。我们的研究小组最近制造了一种重组伪狂犬病病毒（PRV），该病毒以cre重组酶为生存力和GFP报告基因的表达为条件。这种病毒已经成功地用于将输入映射到特定大脑区域的特定神经元群体。为了简单起见，我们将这种方法和相关方法称为PAM，伪狂犬病病毒辅助作图。在本申请中，我们建议进一步开发PAM，以便允许：1）来自相同脑区域中的不同神经元的同时逆行追踪，2）来自不同脑区域中的相似神经元的同时逆行追踪，3）顺行追踪，4）投射到多于一个位点的神经元的识别，5）在这些映射研究中识别的神经元的指定。为了确定该方法的有效性，这些研究将在调节进食的下丘脑回路分析中进行。一旦这些方法得到验证，这些病毒和动物将免费提供给其他对其他神经通路的类似研究感兴趣的研究人员。