Genetic analysis of the adenylate forming domain protein DIP-2 in C. elegans neuronal development

神经元表现出多种多样的形态，有助于连接和功能。神经元形态学是在关键的发育时期建立的，此时细胞骨架结构和重排介导轴突和树突过程的规范、神经元连接的布线以及树突乔木的活性依赖性和独立性阐述。在发育后，神经元的寿命非常长，并且虽然显示出能够促进树突结构的可塑性重塑，但被认为在成年后保持相对稳定的形态。维持神经元形态的分子机制仍然知之甚少。我的实验室感兴趣的是神经元形态是如何建立和维持使用解剖学上简单的神经系统的C。elegans，一种微型蠕虫，作为一个模型系统。我们最近发现，突变dip-2，蠕虫直向同源物的迪斯科相互作用蛋白-2（DIP 2），显示强烈的神经元形态缺陷，在多种类型的神经元，这表明在神经系统发育和维护的核心作用。DIP 2蛋白属于腺苷酸形成结构域蛋白的高度保守但知之甚少的家族。我们的目标与这个发现补助金是了解如何腺苷酸形成域蛋白DIP-2的行动，以维持神经元形态。首先，我们建议采取公正的“基因/蛋白质发现”的方法来识别基因突变，显示DIP-2样神经元形态缺陷或异常DIP-2蛋白定位（目的1A）和DIP-2蛋白复合物的新组件在神经元（目的1B），第二，表征所产生的基因/蛋白质候选人的DIP-2介导的神经元维护（目的2）的直接作用。这些目标的成功完成应该会导致对神经元如何在正常衰老过程中保持其形态的基本见解。

Neurons exhibit a diverse array of morphologies that contribute to both connectivity and function. Neuronal morphology is established during critical developmental periods when cytoskeletal structures and rearrangements mediate the specification of axon and dendritic processes, wiring of neuronal connections and the activity-dependent and independent elaboration of dendritic arbours. Post-developmentally, neurons are very long lived and, while shown to be capable of plasticity-promoting remodeling of dendritic structures, are thought to maintain relatively stable morphologies over adulthood. The molecular mechanisms that maintain neuronal morphology remain poorly understood.*******My lab is interested in how neuronal morphology is established and maintained using the anatomically simple nervous system of C. elegans, a microscopic worm, as a model system. We have recently found that mutations in dip-2, the worm orthologue of Disco Interacting Protein-2 (DIP2), display strong neuronal morphology defects in multiple types of neurons suggesting a central role in both nervous system development and maintenance. DIP2 proteins belong to a highly conserved but poorly understood family of adenylate forming domain proteins. Our objective with this Discovery Grant is to understand how the adenylate forming domain protein DIP-2 acts to maintain neuronal morphology. First, we propose to undertake unbiased ‘gene/protein discovery' approaches to identify mutations in genes that display dip-2-like neuronal morphology defects or aberrant DIP-2 protein localization (Aim 1A) and new components of DIP-2 protein complexes in neurons (Aim 1B) and second, characterize the resulting genes/protein candidates for direct roles in DIP-2 mediated neuronal maintenance (Aim 2). Successful completion of these aims should lead to fundamental insight into how neurons maintain their morphology during the normal aging process.***