New Genetic Tracers for Concurrent Labeling of Multiple Gustatory Neural Pathways

用于同时标记多个味觉神经通路的新基因示踪剂

基本信息

批准号：
7982415
负责人：
ICHIRO MATSUMOTO
金额：
$ 14.44万
依托单位：
MONELL CHEMICAL SENSES CENTER
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-15 至 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7982415
关键词：
Affect Animals Antibodies Area Binding Cells Chemicals Code Computer Simulation Cultured Cells Engineering Enhancers Evaluation FLAG peptide Future Genes Genetic Grant In Vitro Label Lectin Mediating Methods Modality Molecular Molecular Conformation Mus Nervous system structure Neural Pathways Neuraxis Neurosciences Research Oligosaccharides Pathway interactions Peptides Phospholipase C Plant Lectins Plasmids Protein Engineering Proteins Receptor Cell Research Research Personnel Screening procedure Signal Transduction Structure System Taste Perception Testing Tracer Transgenic Animals Transgenic Mice Virus Wheat Germ Agglutinins Work barley lectin base comparative efficacy enhanced green fluorescent protein immunocytochemistry in vitro activity in vivo mRNA Expression molecular dynamics neural circuit promoter protein distribution public health relevance relating to nervous system research study somatosensory tool

项目摘要

DESCRIPTION (provided by applicant): Although in recent decades our understanding of the molecular mechanisms of taste reception has advanced, how the taste information is transmitted and coded in the nervous system remains unclear. Our long-term objective is to understand the coding system of gustatory information, from taste receptor cells to the central nervous system. In this proposal, we aim to develop genetic tools that will greatly contribute to our understanding by concurrently elucidating precise gustatory neural pathways within the same animal, and then use these tools to test our hypothesis that gustatory information for different taste modalities is transmitted via specific ascending neural pathways. One well-known genetic tracer is wheat germ agglutinin (WGA), which was developed ten years ago. In the research we propose here, we will engineer various WGAs that integrate specific tags such as Myc peptide, FLAG peptide, and enhanced green fluorescent protein, and will evaluate their tertiary structure by molecular dynamic simulation. We will evaluate the resultant engineered WGAs in terms of their intracellular stability and lectin activity in vitro. We will then induce their expression in mice in a subset of taste receptor cells under the control of promoters/enhancers of taste reception-related genes and evaluate their utility in vivo. From these studies, we expect to develop a minimum of two tagged WGA- based genetic tracers that are available to label multiple gustatory pathways concurrently and distinguishably. These tracers should be of great use not only in our future gustatory research but in studies of olfactory and somatosensory systems and many other areas of neuroscience research that study the genetics of interconnected pathways. PUBLIC HEALTH RELEVANCE: This proposed research aims to develop new genetic tools to label multiple neural circuitries concurrently and distinguishably. These tools will contribute to elucidating relationships between multiple neural pathways for different taste modalities. They will also contribute to other neuroscience research, including studies of olfactory and somatosensory systems.

描述（由申请人提供）：尽管近几十年来，我们对味觉接收的分子机制的理解已经提高了，但如何在神经系统中传播和编码味觉信息尚不清楚。我们的长期目标是了解从味觉受体细胞到中枢神经系统的味觉信息的编码系统。在这项建议中，我们旨在开发遗传工具，这些工具将通过同时阐明同一动物内的精确味觉神经途径，从而极大地有助于我们的理解，然后使用这些工具来测试我们的假设，即通过特定的上升神经途径传播了不同味道模式的味觉信息。一位著名的遗传示踪剂是十年前开发的小麦胚芽凝集素（WGA）。在我们在这里提出的研究中，我们将设计各种WGA，以整合特定标签，例如MYC肽，FLAG肽和增强的绿色荧光蛋白，并通过分子动态模拟评估其高等教育结构。我们将根据体外的细胞内稳定性和凝集素活性来评估所得工程的WGA。然后，我们将诱导它们在与味觉接收相关基因的启动子/增强子的控制下的味觉受体细胞子集中的小鼠中的表达，并评估它们在体内的效用。从这些研究中，我们期望至少开发两个标记的基于WGA的遗传示踪剂，这些示踪剂可用于同时且区分的多个阵风途径。这些示踪剂不仅在我们的未来味觉研究中，而且在研究互连途径的遗传学的神经科学研究的许多其他领域以及许多其他神经科学研究领域都应使用。公共卫生相关性：这项拟议的研究旨在开发新的遗传工具，以同时且区分地标记多个神经电路。这些工具将有助于阐明多个神经途径之间的关系，以实现不同的口味方式。它们还将为其他神经科学研究做出贡献，包括对嗅觉和体感系统的研究。