Molecular Mechanism of C. elegans gonadogenesis

线虫性腺发生的分子机制

• 批准号: 6804338
• 负责人: JENNIFER A MISKOWSKI
• 金额: $ 19.3万
• 依托单位: UNIVERSITY OF WISCONSIN LA CROSSE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-15 至 2007-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6804338
• 关键词: Caenorhabditis elegans; developmental genetics; gene expression; gene mutation; gonads; green fluorescent proteins; helminth genetics; histogenesis; mutant; phenotype; proteins; reporter genes; reproductive development

DESCRIPTION (provided by applicant): The formation of a complex organ requires multiple developmental processes that must each be properly executed and also coordinated with one another. Investigating the cellular and molecular basis of these 3rocesses provides important insight into normal developmental mechanisms that will expand our understanding of developmental defects and contribute to progress in culturing tissues and organs for medical use. Furthermore, as many diseases result from a normal cellular process gone awry, basic developmental biology research provides key insights into disease initiation, progression, and potential treatments. This proposal is focused on elucidating the role of the Caenorhabditis elegans NUD-1 protein and functionally-related proteins in the development of a specific organ, the gonad. The gonad is a well established model system for studying organogenesis, however, many features of gonad formation are not understood. NUD-1 is a well conserved, but novel protein that provides an inroad into less understood aspects of C. elegans gonadogenesis. First, the gonadal phenotype associated with depletion of NUD-1 will be determined using two distinct approaches. NUD-1 knock-down mutants will be generated using the reverse genetic approach of RNA-mediated interference (RNAi) and characterized using cell-type specific markers. To allow study of the primary gonadal defect associated with loss of NUD-1, a genetic screen will be performed in an attempt to obtain a heritable nud-1 mutation. The second aim is to characterize the gonadal expression pattern of NUD-I. Again, a two-pronged approach will be employed and both NUD- 1::GFP expression constructs and a NUD-l-specific antibody will be generated and analyzed. Finally, other mutations with a Nud phenotype will be identified in the genetic screen and characterized. Experiments will be performed to determine if the newly identified loci function with nud-1, or with each other, in gonad development. Overall, the proposed work is designed to increase our knowledge of mechanisms used in organ formation and to shed light on the means by which NUD-1 exerts its effects.

描述（由申请人提供）：复杂器官的形成需要多个发育过程，每个过程都必须正确执行并且相互协调。研究这三个过程的细胞和分子基础为正常发育机制提供了重要的见解，这将扩大我们对发育缺陷的理解，并有助于培养医用组织和器官的进展。此外，由于许多疾病是由正常细胞过程出错引起的，基础发育生物学研究为疾病的发生、进展和潜在治疗提供了重要见解。该提案的重点是阐明秀丽隐杆线虫 NUD-1 蛋白和功能相关蛋白在特定器官（性腺）发育中的作用。性腺是研究器官发生的成熟模型系统，然而，性腺形成的许多特征尚不清楚。 NUD-1 是一种高度保守但新颖的蛋白质，它为线虫性腺发生的鲜为人知的方面提供了一个新的途径。首先，将使用两种不同的方法来确定与 NUD-1 消耗相关的性腺表型。 NUD-1 敲低突变体将使用 RNA 介导的干扰 (RNAi) 的反向遗传方法产生，并使用细胞类型特异性标记进行表征。为了研究与 NUD-1 缺失相关的原发性性腺缺陷，将进行基因筛查，试图获得可遗传的 nud-1 突变。第二个目的是表征 NUD-I 的性腺表达模式。再次，将采用双管齐下的方法，并且将生成并分析NUD-1::GFP表达构建体和NUD-1特异性抗体。最后，具有 Nud 表型的其他突变将在遗传筛选中被鉴定并表征。将进行实验以确定新鉴定的基因座在性腺发育中是否与 nud-1 一起发挥作用，或者彼此之间是否发挥作用。总体而言，拟议的工作旨在增加我们对器官形成机制的了解，并阐明 NUD-1 发挥其作用的方式。