A high-throughput sequencing and imaging approach to understand the functional basis of olfaction

用于了解嗅觉功能基础的高通量测序和成像方法

基本信息

批准号：
10468179
负责人：
Dinu Florentin ALBEANU
金额：
$ 106.53万
依托单位：
COLD SPRING HARBOR LABORATORY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-11 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10468179
关键词：
Affinity Age Animals Area Bar Codes Binding Brain Cells Chemicals Code Cognition Cognitive deficits Data Devices Fire - disasters Fluorescence Food Foundations Gases Goals High-Throughput DNA Sequencing High-Throughput Nucleotide Sequencing Human In Situ Individual Label Ligands Link Location Logic Maps Methods Modality Molecular Molecular Biology Multiplexed Analysis of Projections by Sequencing Mus Neurons Nose Odorant Receptors Odors Olfactory Epithelium Olfactory Pathways Optics Output Pattern Process Property Quality of life RNA Receptor Gene Research Resolution Sampling Sensory Smell Perception Specificity Specimen Stimulus Structure Surface Technology Therapeutic Tissues Transcript Vision Work disease diagnostic experimental study imaging approach in situ sequencing in vivo in vivo imaging in vivo optical imaging innovation insight interdisciplinary approach multiphoton imaging neural circuit neurophysiology new technology next generation next generation sequencing olfactory bulb olfactory bulb glomeruli olfactory sensory neurons olfactory stimulus optical imaging response statistics tool transcriptome sequencing two-photon

项目摘要

PROJECT SUMMARY We propose to develop a strategy for understanding olfactory coding by linking the molecular identity of odorant receptors (OR) to their odor sensitivities in vivo and discovering the logic of neural circuits that process smell. Unlike for other sensory modalities (i.e. vision, audition), it is not understood what properties of odorants are important for olfaction, how these properties are processed by olfactory neural circuits, and how odorant receptor genes have evolved to optimize such an encoding. The relationship between odorant structure (chemical space), the sequence of odorant receptors, the underlying spatial-temporal patterns of activity in the brain (neuronal space) and the perceived odor quality (perceptual space) has been elusive. An efficient method for connecting the olfactory sensory spaces will have a paradigm-shifting effect on olfactory research. Our multidisciplinary approach will use cutting edge next-generation sequencing technologies (FISSEQ, MAPseq and RNAseq) together with functional widefield fluorescence and two photon imaging in vivo to define the functional properties of olfactory sensory neurons that express defined odorant receptors (ORs), to discover their connections to individual glomeruli olfactory bulb (OB), second order OB output (mitral/tufted) cells and map their projection statistics to the downstream olfactory processing brain areas. Using these tools, we will map the identity and spatial layout of all 3,500 glomeruli in the mouse olfactory bulb according to the OR types from which they receive inputs. We will further link the molecular identity of ORs to their glomerular responses to hundreds of odorants (>500) in the form of OR/odorant binding affinity matrices across hundreds of glomeruli (~500) that are optically accessible in vivo. In the same samples, we will track thousands (>1,000/experiment) of individual olfactory bulb projections to their input glomeruli and their target brain areas by RNA-barcoding in relation to their tuning to odorants via multiphoton imaging in vivo. Our approach will bridge the gap between the molecular biology of ORs and neurophysiology and will usher in a new era of understanding the functional basis of olfaction. It will allow unprecedented resolution and throughput for determining OR-ligand interactions across hundreds of odorants, and the connectivity of tens of thousands of single neurons at once in a single specimen. The data obtained will enable the study of OR- ligand interactions, relate the chemical identity of odorants to olfactory perception, and the construction of artificial nose devices for immediate biomedical applications, including disease diagnostics.

项目概要我们建议通过链接嗅觉编码的分子身份来开发一种理解嗅觉编码的策略气味受体（OR）对体内气味敏感性的研究，并发现处理气味的神经回路的逻辑闻。与其他感官方式（即视觉、听觉）不同，我们不了解气味剂的特性对于嗅觉很重要，嗅觉神经回路如何处理这些特性，以及气味如何受体基因已经进化以优化这种编码。气味结构之间的关系（化学空间），气味受体的序列，潜在的时空活动模式大脑（神经元空间）和感知气味质量（感知空间）一直难以捉摸。一个高效的连接嗅觉感觉空间的方法将对嗅觉研究产生范式转换的影响。我们的多学科方法将使用尖端的下一代测序技术（FISSEQ、 MAPseq 和 RNAseq）与功能性宽场荧光和体内双光子成像一起定义表达特定气味受体（OR）的嗅觉感觉神经元的功能特性，发现它们与单个肾小球嗅球 (OB) 的连接，二阶 OB 输出（二尖瓣/簇状）细胞并将其投影统计数据映射到下游嗅觉处理大脑区域。使用这些工具，我们将根据以下模型绘制小鼠嗅球中所有 3,500 个肾小球的身份和空间布局它们接收输入的 OR 类型。我们将进一步将 OR 的分子特性与其肾小球联系起来以 OR/气味结合亲和力矩阵的形式对数百种气味剂 (>500) 做出反应体内可光学接近的肾小球（〜500）。在相同的样本中，我们将跟踪数千个（>1,000/实验）个体嗅球投射到其输入肾小球和目标大脑区域通过RNA条形码编码，通过体内多光子成像调节气味剂。我们的方法将弥合 OR 分子生物学和神经生理学之间的差距，并将开创了理解嗅觉功能基础的新时代。它将提供前所未有的分辨率和用于确定数百种气味剂之间的 OR 配体相互作用的吞吐量，以及数十种气味剂的连接性一个样本中同时存在数千个单个神经元。获得的数据将使 OR-的研究成为可能配体相互作用，将气味剂的化学特性与嗅觉感知联系起来，以及用于直接生物医学应用（包括疾病诊断）的人造鼻装置。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Deconstructing Odorant Identity via Primacy in Dual Networks.

通过双网络中的首要地位解构气味身份。

DOI：
发表时间：
2019
期刊：
影响因子：
2.9
作者：
Kepple, Daniel R;Giaffar, Hamza;Rinberg, Dmitry;Koulakov, Alexei A
通讯作者：
Koulakov, Alexei A

Mosaic representations of odors in the input and output layers of the mouse olfactory bulb.

小鼠嗅球输入层和输出层气味的马赛克表示。

DOI：
发表时间：
2019
期刊：
影响因子：
25
作者：
Chae, Honggoo;Kepple, Daniel R;Bast, Walter G;Murthy, Venkatesh N;Koulakov, Alexei A;Albeanu, Dinu F
通讯作者：
Albeanu, Dinu F