Behavioral and Neural Deterioration in Drosophila

该提案描述了理解年龄依赖性行为和神经退行性变模型的遗传、分子和细胞基础的第一步。这与人类健康和生活质量高度相关，因为先进的医学现在允许许多人在严重的行为和认知缺陷下生存多年。这些研究的切入点是最近的观察结果，即果蝇kelch基因的突变导致交配成功率、性感受性和身体控制方面的年龄依赖性缺陷，而生存能力没有显著降低。这些研究的相关性增加了神经系统表达的同源物的Kelch在人类的鉴定。该项目的长期目标是了解与Kelch突变相关的神经和行为缺陷的细胞基础，将其与Kelch的分子作用联系起来，使用Kelch突变的适当操作来筛选与Kelch相互作用的其他蛋白质以维持神经功能，并模拟野生型和突变型人类Kelch样蛋白的潜在作用。在这一试点项目中，将特别强调为更宏伟的长期目标奠定必要的基础。分子定位的无义和错义突变将仔细表征多种行为表型的严重程度和时间进程。 这将决定可能的kelch表型阵列和定义的蛋白质结构域的改变的后果。 在表征年龄依赖性行为表型的同时，将检查CNS、神经肌肉系统和视觉系统中的年龄相关异常，包括CNS整体和神经元或神经胶质的定义子集。CNS或外周神经系统改变的表征将与通过RNA杂交和Kelch特异性抗体确定的Kelch表达模式相关。将构建用于后续研究的试剂，包括在动物或培养物中靶向表达Kelch、GFP-Kelch和相应的人蛋白，以及检测小群体Kelch突变细胞。

This proposal describes the first steps in understanding the genetic, molecular and cellular basis of one model of age-dependent behavioral and neural degeneration. This has become highly relevant to human health and quality of life as advanced medicine now allows many people to survive for years with significant behavioral and cognitive deficits. The entree into these studies is the recent observation that mutations in the Drosophila kelch gene lead to age-dependent deficits in mating success, sexual receptivity, and body control without significantly reduced viability. The relevance of these studies is increased by the identification of nervous-system-expressed homologs of kelch in humans. The long term goal of this project is to understand the cellular basis of neural and behavioral defects associated with kelch mutations, to link this to the molecular action of Kelch, to use appropriate manipulations of kelch mutations to screen for addition proteins interacting with Kelch in maintenance of neural function, and to model the potential action of wild type and mutant versions of the human Kelch-like proteins. In this pilot project special emphasis will be given to laying the necessary foundation for the more ambitious long term goals. Molecularly mapped nonsense and missense mutations will be carefully characterized for severity and time course of multiple behavioral phenotypes. This will determine the array of possible kelch phenotypes and the consequences of alterations in defined protein domains. In parallel with the characterization of age- dependent behavioral phenotypes, age-associated abnormalities in the CNS, neuromuscular system, and visual system will be examined, both in the CNS as a whole and in defined subsets of neurons or glia. Characterization of alterations in the CNS or peripheral nervous system will be correlated with expression patterns for Kelch as determined by RNA hybridization and Kelch-specific antibodies. Reagents for later studies, including targeted expression of Kelch, GFP-Kelch and corresponding human proteins in animals or in culture, and examination of small groups of Kelch mutant cells will be constructed.