AN ANALYSIS OF THE ROLE OF THE CELEGANS SECRETOME ON NEURAL DEVELOPMENT BY RNAI

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 神经系统中的细胞广泛使用细胞间信号线索来适当地延伸神经突起，导航和识别靶细胞，形成和维持突触联系。从整个基因组中预测分泌蛋白质的能力导致了这样的结论：可能有多达三分之一的编码蛋白质被分泌到细胞外空间。考虑到神经系统的复杂性，理所当然地，这些分泌的蛋白质中的许多都在神经系统的模式中起作用。 我们已经从线虫基因组中建立了一个预测分泌蛋白的文库。我们将采用一种系统的方法，使用RNA干扰(RNAi)来敲除所有7,460个预测的分泌蛋白，并评估这种RNAi对神经发育的多个方面的影响。将进行初步表征，以确定对神经元模式有影响的分子，并将对这些分子进行二级定量分析。我们正在使用一种合成的致命筛选方法来首先表征这些分子，然后我们将检查突变体中显示出效应的基因的突起和突触形态。 了解神经系统是如何形成模式的一直是神经生物学的目标。大量的研究已经证明了细胞外蛋白在神经元发育的各个方面的重要性。系统地评估所有分泌蛋白质功能丧失的影响的能力直到最近才变得可行。这将为开发中一个极其复杂的部分提供许多新颖的见解。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Cells in the nervous system use intercellular signaling cues extensively to properly the extend neurites, navigate to and recognize target cells and form and maintain synaptic contacts. The ability to predict secreted proteins from whole genomes has led to the conclusion that perhaps as much as one third of all proteins encoded are secreted into the extracellular space. Given the complexity of nervous systems it stands to reason that many of these secreted proteins act in the patterning of the nervous system. We have generated a library of predicted secreted proteins from the C. elegans genome. We will take a systematic approach using RNA interference (RNAi) to knockdown of all 7,460 predicted secreted proteins and evaluate the effect of this RNAi on multiple aspects of neural development. A preliminary characterization will be done to identify molecules that have an effect on neuronal patterning, and a secondary level quantitative analysis will be done on those molecules. We are using a synthetic lethal screening approach to characterize these molecules first, and then we will examine the outgrowth and synaptic morphology in mutants for genes that demonstrate an effect. Understanding how the nervous system is patterned has long been a goal of neurobiology. A multitude of studies have demonstrated the importance of extracellular proteins on all aspects of neuronal development. The ability to systematically evaluate the effect of loss of function of all secreted proteins has only recently become available. This will provide many novel insights into an extremely complicated part of development.