Imaging Taste in Ensembles of Afferent Neurons

传入神经元集合中的味觉成像

基本信息

批准号：
8512700
负责人：
Nirupa Chaudhari
金额：
$ 18.17万
依托单位：
UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-20 至 2014-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8512700
关键词：
Action Potentials Afferent Neurons Animals Axon Biological Assay Bite Brain Cataloging Catalogs Cell Count Cell Nucleus Cells Chemicals Code Communities Data Databases Development Diet Dyes Entropy Enzymes Exploratory/Developmental Grant Fatty acid glycerol esters Fiber Fluorescence Functional Imaging Ganglia Garlic General Practitioners Genetically Engineered Mouse Glossopharyngeal nerve structure Human Image Individual Ion Channel Label Laser Scanning Confocal Microscopy Lasers Life Maps Measures Methods Molecular Molecular Profiling Mus Nerve Neurons Neurotransmitters Nucleus solitarius Oral Oral cavity Organ Outcome Pathway interactions Peripheral Process Proteins Quality of life Rattus Reporter Research Resolution Reverse Transcriptase Polymerase Chain Reaction Scanning Sensory Sensory Ganglia Signal Transduction Specialist Stimulus Structure of geniculate ganglion System Taste Bud Cell Taste Buds Taste Perception Techniques Transgenic Organisms Work chorda tympani combinatorial ganglion cell high reward high risk hindbrain insight novel novel strategies receptor research study response salt sensitive transcription factor

项目摘要

DESCRIPTION (provided by applicant): This multi-PI R21 proposal describes a high-risk, high-impact project that introduces a novel method for recording taste-evoked activity in gustatory afferent neurons. The work represents a major new direction for the PIs as well as for the chemical senses community, and thus is ideally suited for the R21 mechanism. The project will introduce a novel recording technique to produce new data on the taste sensitivities and transmitter systems of large ensembles of geniculate ganglion neurons. We will employ a new strain of genetically-engineered mice that express a fluorescent functional reporter, GCaMP3, selectively in sensory (including gustatory) ganglion neurons. We propose to develop a new recording technique to image taste-evoked activity in fluorescent geniculate ganglion neurons in anesthetized mice. Geniculate neurons will be imaged with scanning laser confocal microscopy and changes in fluorescence quantified to simultaneously measure activity in large ensembles of neurons with single cell resolution. Ganglion cells will be excited by prototypic sweet, bitter salty, sour, umami and fat taste stimuli, delivered in the oral cavity. We will measure the breadth of tuning, concentration-response relations and entropy for gustatory geniculate ganglion cells (Aim 1). Functionally characterized neurons will be isolated and single cell RT-PCR will be carried out to examine the neurotransmitter systems employed by functionally distinct taste neurons (Aim2). Successful completion of the two aims will have 4 outcomes: first, we will have developed a completely new method for recording afferent sensory activity in large numbers of gustatory neurons~ second, we will accumulate a large database of response profiles for gustatory afferent axons, providing a comprehensive catalog of their breadth of tuning and entropies~ third, we will determine whether different classes of sensory neurons ("generalists"~ "specialists"~ sweet-, sour-sensing, etc.) have different molecular expression profiles, such as distinctive transmitter systems, transcription factors, and so forth~ and finally, whether there i a unique class of fat-sensing geniculate ganglion neurons. The data will have implications for how taste is coded by sensory afferents (e.g., labeled line vs, combinatorial coding) and will tremendously increase our understanding about how sensory neurons process gustatory signals.

描述（由申请人提供）：此多PI R21提案描述了一个高风险，高影响力的项目，该项目引入了一种新颖的方法，用于记录味觉传入神经元中的味觉引起的活动。这项工作代表了PI和化学感官社区的主要新方向，因此非常适合R21机制。该项目将引入一种新型的录音技术，以产生有关吉属神经节神经元大集合的味觉敏感性和发射器系统的新数据。我们将采用一种新的遗传工程小鼠菌株，该菌株在感觉（包括味觉）神经节神经元中有选择地表达荧光功能报告GCAMP3。我们建议开发一种新的录音技术，以在麻醉小鼠的荧光基因神经节神经元中成像味道诱发的活性。将使用扫描激光共聚焦显微镜成像，并在具有单细胞分辨率的大型神经元的大团体中测量荧光变化，并在荧光中成像。在口腔中传递的原型甜，酸味，酸，鲜味和脂肪味刺激将激发神经节细胞。我们将测量广度调谐，浓度 - 响应关系和味道基因神经节细胞的熵（AIM 1）。功能表征的神经元将被隔离，将进行单细胞RT-PCR，以检查功能上不同味道神经元采用的神经递质系统（AIM2）。这两个目标的成功完成将有4个结果：首先，我们将开发一种全新的方法来记录大量味觉神经元的传入感觉活动〜其次，我们将积累大量的响应数据库，用于味觉传入的轴突，以提供全面的响应性神经元，从酸味等）具有不同的分子表达曲线，例如独特的发射器系统，转录因子等，最后，是否有I唯一的一类脂肪感应的神经节神经元。这些数据将对感官传入的编码（例如标记的线与组合编码）的编码有影响，并将极大地增加我们对感觉神经元如何处理味道信号的理解。