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

要了解细胞是如何发育或发挥功能的，重要的是研究不同个体的等值细胞。 比较突变型和野生型个体的细胞命运可以揭示基因是如何调节的 发展。同样，将来自一种动物的关于神经元结构的信息与功能 对另一个神经元活动的记录可以揭示该神经元对电路的贡献。比较信息 跨动物需要通过细胞形状、位置、基因表达和基因表达的某种组合来识别细胞 血统。然而，在复杂的组织中识别细胞命运的方法是简单的，集中在一个或一个小的 一次只有几种细胞类型，因此很难评估整个群体的行为。我们建议 开发多路复用3D多色荧光标记及相关硬件和软件，以 在活体动物中自动识别细胞。在目标1中，我们将在线虫C. 优雅是因为它不变的发育谱系和已知的每个神经元的连接提供了强大的 用于构建、测试和验证此方法的平台。在目标2中，我们将应用这一点来理解 在发育突变中整个有机体的命运变化。最后，在目标3中，我们将使用我们的 标记法识别成人神经环(脑)中的所有神经元，并将其与全脑相结合 首次将每个神经元的活动映射到接线图上的成像方法。

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.