Behavioral and Neural Deterioration in Drosophila

这一建议描述了理解的遗传，分子和细胞基础的一个模型的年龄依赖的行为和神经退化的第一步。这与人类健康和生活质量高度相关，因为先进的医学现在可以让许多患有严重行为和认知缺陷的人存活多年。这些研究的开端是最近观察到的果蝇kelch基因突变导致交配成功、性接受和身体控制方面的年龄依赖性缺陷，而不会显著降低生存能力。这些研究的相关性增加了识别人类kelch的神经系统表达同源物。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，使用适当的kelch突变操作来筛选与kelch相互作用以维持神经功能的附加蛋白，并模拟野生型和突变版本的人类kelch样蛋白的潜在作用。在这个试点项目中，将特别强调为更宏伟的长期目标奠定必要的基础。分子映射的无义和错义突变将仔细表征的严重性和时间过程的多种行为表型。这将决定可能的kelch表型阵列和定义的蛋白质结构域改变的后果。在对年龄依赖性行为表型进行表征的同时，将对中枢神经系统、神经肌肉系统和视觉系统中与年龄相关的异常进行检查，包括中枢神经系统作为一个整体以及特定的神经元或神经胶质亚群。中枢神经系统或周围神经系统改变的特征将与RNA杂交和Kelch特异性抗体确定的Kelch表达模式相关。将构建用于后期研究的试剂，包括Kelch、GFP-Kelch和相应的人蛋白在动物或培养中的靶向表达，以及对Kelch突变细胞小群体的检测。

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.