Multicolor labeling for cell identification in the C. Elegans Nervous system

用于线虫神经系统细胞识别的多色标记

基本信息

批准号：
9456323
负责人：
John Isaac Murray
金额：
$ 25.75万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9456323
关键词：
Ablation Adult Aging Alpha Cell Animals Behavior Brain Brain imaging Caenorhabditis elegans Calcium Cell Count Cell Lineage Cell Proliferation Cell Shape Cell physiology Cells Collection Color Complex Computer software Computing Methodologies Databases Defect Development Embryo Embryonic Development Embryonic Nervous System Fluorescence Functional Imaging Future Gene Expression Genes Genetic Head Image Individual Investigation Label Location Locomotion Maps Measurement Measures Methods Modeling Morphology Motor Nematoda Nerve Nervous System Physiology Nervous system structure Neurons Organism Pathway interactions Pattern Phenotype Population Population Study Positioning Attribute Process Property Proteins Resolution Role Specificity Structure Techniques Technology Testing Three-Dimensional Imaging Time Tissues Transgenic Organisms blastomere structure cell motility cell type connectome experimental study gene function imaging Segmentation imaging modality imaging platform improved mutant nerve stem cell neural circuit optical imaging progenitor promoter relating to nervous system tool

项目摘要

To understand how cells develop or function, it is important to study equivalent cells across individuals. Comparing a cell's fate between mutant and wild-type individuals can reveal how a gene regulates development. Similarly, combining information about a neuron's structure from one animal with functional recordings of its activity in another can reveal that neuron's contribution to a circuit. Comparing information across animals requires identifying cells by some combination of cell shape, position, gene expression and lineage. However, methods to identify cell fates in complex tissues are simplex, focusing on one or a small number of cell types at a time, making it hard to assess the behavior of the full population. We propose to develop multiplexed 3D multicolor fluorescent labeling and associated hardware and software to automate cell identification in live animals. In Aim 1 we will develop this approach in the nematode C. elegans because its invariant developmental lineage and known wiring of every neuron provides a powerful platform on which to build, test and validate this approach. In Aim 2 we will apply this to understand patterns of fate transformations across the entire organism in developmental mutants. Finally in Aim 3, we will use our labeling method to identify all neurons in the adult nerve ring (brain), and combine this with whole-brain imaging methods to map for the first time activity of every neuron onto the wiring diagram.

要了解细胞的发展或功能，重要的是研究跨个体的等效细胞。比较突变体和野生型个体之间的细胞的命运可以揭示基因如何调节发展。同样，将一只动物的神经元结构的信息与功能相结合其活动中其活动的记录可以表明神经元对电路的贡献。比较信息整个动物都需要通过细胞形状，位置，基因表达和血统。但是，鉴定复杂组织中细胞命运的方法是单纯形，重点是一个或小一次性细胞类型的数量，使得很难评估全部人群的行为。我们建议开发多路复用的3D多色荧光标签以及相关的硬件和软件在活动物中自动化细胞鉴定。在AIM 1中，我们将在线虫C中开发这种方法。秀丽隐杆线虫是因为它不变的发育谱系和每个神经元的已知接线提供了强大的功能可以在其中构建，测试和验证这种方法的平台。在AIM 2中，我们将使用此操作来了解整个生物体中发育突变体的命运转变。终于在AIM 3中，我们将使用我们的标记方法以识别成年神经环（大脑）中的所有神经元，并将其与全脑结合成像方法将每个神经元的首次活动映射到接线图上。