Molecular Multi-Species Approach for Trans-Synaptic Labeling of Neural Circuits
神经回路跨突触标记的分子多物种方法
基本信息
- 批准号:10009743
- 负责人:
- 金额:$ 273.18万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2020
- 资助国家:美国
- 起止时间:2020-08-01 至 2024-07-31
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
PROJECT SUMMARY/ABSTRACT
It is estimated that the human brain contains an overwhelming 1015 synapses, structures essential for the normal
functioning of neural circuits. Our knowledge of the connections that form these critical signaling sites, in even
the simplest vertebrate nervous systems, is sorely lacking. Thus, a stated goal of this BRAIN initiative is to
“develop and validate novel tools to facilitate the detailed analysis of complex circuits and provide insights into
cellular interactions that underlie brain function”. This multi-PI collaborative project precisely addresses this goal.
It takes advantage of a powerful genetic technique, trans-Tango, that directs signaling across synapses to
identify both pre-synaptic neurons and their specific post-synaptic targets. The overall objectives of the proposed
experiments are three-fold: First, we will adapt the trans-Tango anterograde trans-synaptic signaling platform,
which was initially established and successfully implemented in the Drosophila model, to a vertebrate brain - that
of the zebrafish. The zebrafish is the organism of choice because of the ability to assay trans-Tango components
efficiently from injections of plasmid constructs into 1-cell embryos, and the ease and rapidity of generating
transgenic animals to activate trans-Tango in defined neuronal populations. Second, we will independently and
rigorously validate the neural connections revealed by trans-Tango as functional synaptic connections,
capitalizing on optogenetics, imaging techniques, and advanced microscopy methods. Owing to its transparency,
the larval zebrafish is ideally suited to verify synaptic connectivity in vivo using optical approaches. Third, we will
develop a new retrograde version of trans-Tango, which will allow identification of the pre-synaptic input of given
post-synaptic neurons. The modularity of trans-Tango permits efficient reconfiguration and optimization of the
system for accurate circuit
map
ping. The “retro-Tango” version will first be applied to Drosophila, building upon
lessons learned from the establishment of trans-Tango and, once optimal, introduced to the zebrafish nervous
system. By assembling the proposed genetic toolkit for anterograde and retrograde trans-synaptic tracing in both
invertebrate and vertebrate nervous systems, we expect these techniques to become widely used by the
neuroscience community and applied to additional experimental models. The strengths of this proposal are the
innovative strategies used to
map
neural connectivity, the compelling preliminary data, and the unique and
complementary expertise in molecular genetics, circuit neuroscience and microscopy design that the
collaborating researchers bring to the project.
项目摘要/摘要
据估计,人脑含有压倒性的1015个突触,对正常必不可少的结构
神经回路的功能。我们对形成这些关键信号站点的连接的了解,甚至
最简单的脊椎动物神经系统非常缺乏。这是这项大脑计划的一个明确的目标是
“开发和验证新颖的工具,以促进复杂电路的详细分析,并为
大脑功能构成的细胞相互作用”。这个多PI协作项目正好解决了这一目标。
它利用了一种强大的遗传技术,即Trans-Tango,该技术将跨突触的信号引导到
确定突触前神经元及其特定的突触后靶标。提议的总体目标
实验是三倍:首先,我们将适应反式tango tango tango theRofrade反式突触信号平台,
最初是在果蝇模型中建立并成功实现的,脊椎动物的大脑 -
斑马鱼。斑马鱼是选择的生物,因为能够测定反式tango组件
有效地从注射质粒构建到1细胞胚胎中,以及产生的易度和速度
转基因动物激活定义的神经元种群中的thango。其次,我们将独立并
严格验证Trans-Tango揭示的神经连接是功能突触连接,
利用光遗传学,成像技术和晚期显微镜方法。由于其透明度,
幼虫斑马鱼非常适合使用光学方法在体内验证合成连通性。第三,我们会的
开发新的逆行版本的Trans-Tango,该版本将允许识别给定的突触前输入
突触后神经元。反式tango的模块化允许有效的重新配置和优化
精确电路系统
地图
ping。 “ retro-tango”版本将首先应用于果蝇,建立在果蝇上
从Trans-Tango的建立中学到的经验教训,一旦最佳地引入斑马鱼
系统。通过组装提出的基因工具包
无脊椎动物和脊椎动物神经系统,我们预计这些技术将被广泛使用
神经科学界,并应用于其他实验模型。该提议的优势是
创新策略曾经
地图
神经连通性,引人入胜的初步数据以及独特的和
在分子遗传学,电路神经科学和显微镜设计方面的完全专业知识,
合作研究人员将其带入该项目。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)

暂无数据
数据更新时间:2024-06-01
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