Catecholaminergic Modulation of Taste Circuitry Using Optogenetics

利用光遗传学调节味觉回路的儿茶酚胺能

基本信息

批准号：
8773145
负责人：
JOSEPH B TRAVERS
金额：
$ 23.1万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-06-01 至 2016-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8773145
关键词：
Automobile Driving Binding Brain Stem Cachexia Catecholamines Cations Cells Clinical DNA Discrimination Disease Dopamine Feeding behaviors Frequencies Functional disorder Genes Genetic Glutamate Transporter Glutamates Health Homeostasis Hormonal In Vitro Injection of therapeutic agent Intake Label Lasers Light Membrane Mixed Function Oxygenases Motivation Mus Nature Neural Pathways Neuraxis Neurons Norepinephrine Nucleus solitarius Obesity Optics Oral Outcome Pathway interactions Phenotype Population Population Heterogeneity Preparation Process Prosencephalon Proteins Rattus Relative (related person)Satiation Signal Transduction Site Stimulus Sucrose Synapses System Taste Perception Techniques Testing Therapeutic Tissues Transgenic Mice Viral Vector Virus Visceral Work bariatric surgery feeding glucagon-like peptide glucagon-like peptide 1 high risk homeodomain in vivo mouse model novel optogenetics parabrachial nucleus promoter relating to nervous system research study response transcription factor vector

项目摘要

DESCRIPTION (provided by applicant): This proposal describes a high risk, high impact project using an optogenetic approach to determine how visceral neurons associated with satiey modulate the taste system. We have created a novel viral vector (AAV-9, PRSx8-ChR2(h134r)-mCherry) that expresses channelrhodopsin-2 (Chr2) and mCherry under the control of a Phox2-selective promoter that labels catecholaminergic neurons containing norepinephrine and other non-GABAergic viscerosenory neurons. A second optogenetic approach will target catecholamine neurons more specifically using a transgenic mouse that expresses Cre under the control of a dopamine-β hydroxylase promoter (DβHCre/0) and a cre-dependent virus driving ChR2 expression. Neurons in the caudal brainstem including catecholamine neurons are intimately associated with visceral signaling underlying satiey mechanisms and glucoprivic-induced feeding, but where and how they modulate taste signals is unknown. Although catecholamine neurons are clustered in groups, e.g. A1 & A2, these clusters are embedded in a more heterogeneous population of cells making it difficult to study specific neural pathways. Optogenetic techniques allow defined neuron types to be specifically activated. Neuronal DNA is modified by inserting a gene expressing a light-sensitive channel (protein) capable of depolarizing the neuron membrane. Following neuronal transduction with the genetic construct, optical stimulation with a laser or high-intensity LED tuned to the optimal frequency of the light-sensitive channel will subsequently stimulate and depolarize those neurons genetically defined by the promoter. Recent work suggests that significant interactions between caudal solitary nucleus neurons sensitive to visceral signals and feeding circuits occur through local brainstem pathways including intrasolitary pathways. The outcomes of our proposal will thus (1) provide a novel technical approach to determine how a specific group of visceral-sensitive neurons modulate gustatory neurons and (2) determine if a previously unexplored brainstem intrasolitary pathway contibutes to taste/visceral integration.

描述（由适用提供）：该提案使用光遗传学方法描述了一个高风险，高影响力项目，以确定与Satiey相关的内脏神经元如何调节味觉系统。我们创建了一种新型的病毒载体（AAV-9，PRSX8-CHR2（H134R）-Mcherry），该媒介在控制phox2-选择性启动子的控制下表达ChannelRhopopsin-2（ChR2）和MCHERRY，该启动子标记了catecholamin offeramin offienagic神经元含有非甲肾上腺素和其他非Gababaer neurons and Neururons neurons and neuron Neurons。第二种光遗传学方法将使用转基因小鼠更具体地靶向儿茶酚胺神经元，该小鼠在控制多巴胺-β羟化酶启动子（DβHCRE/0）的控制下表达CRE和驱动CHR2表达的CRE依赖性病毒。包括儿茶酚胺神经元在内的尾脑干中的神经元与阳性机制的内脏信号传导和葡萄糖诱导的喂养密切相关，但是它们在何处以及如何调节味道信号是未知的。尽管儿茶酚胺神经元聚集成组，例如A1和A2，这些簇被嵌入到更异质的细胞中，因此很难研究特定的神经元途径。光遗传技术允许特异性地激活定义的神经元类型。神经元DNA通过插入表达能够分离神经元膜的光敏感通道（蛋白）的基因来改变。在使用遗传构建体的神经元翻译后，使用激光或高强度LED调整为光敏通道的最佳频率的光学刺激将随后刺激并部署由启动子基因定义的那些神经元。最近的工作表明，对内脏信号敏感的尾固体核核神经元与喂养电路之间的显着相互作用通过包括内部途径在内的局部脑干途径发生。因此，我们的提案的结果将（1）提供一种新型的技术方法来确定特定的内脏敏感神经元如何调节味觉神经元，并且（2）确定先前出乎意料的脑干内部内部途径是否具有味觉/内脏整合。