Examination of PTL-1 in mechanosensory neurons of Caenorhabditis elegans

描述（由申请人提供）：本提案的目的是阐明微管相关蛋白PTL-1在秀丽隐杆线虫机械感觉神经元的功能和细胞结构中的作用。高度保守的结构微管相关蛋白家族，其中PTL-1是秀丽隐杆线虫中发现的唯一成员，被认为在发育和成年神经元中具有许多功能，包括调节微管的动力学和作为参与信号转导的蛋白质组装的动态细胞骨架支架。哺乳动物神经元中这些蛋白质的功能障碍与阿尔茨海默病和中风等神经退行性疾病有关，证明了这些蛋白质在神经元功能中的重要性。然而，对于这些蛋白质如何参与健康神经元的这些功能，我们知之甚少。通过利用秀丽隐杆线虫的遗传学优势和成像能力，我们将进一步了解这组细胞骨架蛋白在神经元功能中的作用。我们将使用遗传和行为策略来确定PTL-1与成年线虫机械感觉神经元中发现的特殊微管之间的关系。我们将使用共聚焦激光扫描显微镜和免疫沉淀结合遗传转化技术来揭示PTL-1与这些微管以及机械感觉转导机制组成部分的蛋白质-蛋白质相互作用。通过了解PTL-1如何与这些蛋白质相互作用，我们将更好地了解该蛋白质家族在健康神经元中的更一般作用，以及更好地了解这类蛋白质在神经元功能中的进化意义。我们还将进一步了解由于疾病或损伤导致的这类蛋白质功能障碍的后果。脑卒中损伤和阿尔茨海默病的标志性特征之一是结构微管相关蛋白MAP2和tau的功能障碍。这种功能障碍的后果尚不清楚，因为人们对这类重要的神经元细胞骨架蛋白在健康神经元中的功能知之甚少。该项目将确定该家族成员之一PTL-1在线虫（秀丽隐杆线虫）健康神经元中的功能，作为增加我们对这类蛋白质在正常健康成年神经元中如何工作的理解的一种方式，这将为这些蛋白质在疾病状态下功能障碍对神经元的影响提供重要线索。

DESCRIPTION (provided by applicant): The objective of this proposal is to elucidate the role of the microtubule-associated protein, PTL-1, in the function and cytoarchitecture of mechanosensory neurons from Caenorhabditis elegans. The highly conserved family of structural microtubule- associated proteins, of which PTL-1 is the only member found in C. elegans, have been postulated to serve numerous functions in developing and adult neurons, including regulating the dynamics of microtubules and serving as dynamic cytoskeletal scaffolds for the protein assemblies involved in signal transduction. Dysfunction of these proteins in mammalian neurons has been implicated in neurodegenerative disorders like Alzheimer's disease and stroke, attesting to the importance of these proteins in neuron function. Little is known, however, about how these proteins are involved in these functions in healthy neurons. By utilizing the strengths of the genetics and the imaging capabilities of C. elegans, we will further our understanding of the role of this group of cytoskeletal proteins in neuron function. We will use genetic and behavioral strategies to determine the relationship between PTL-1 and the specialized microtubules found in mechanosensory neurons in adult nematodes. We will use confocal laser scanning microscopy and immunoprecipitation combined with genetic transformation techniques to reveal the protein-protein interactions of PTL-1 with these microtubules, as well as with components of the mechanosensory transduction machinery. By understanding how PTL-1 interacts with these proteins, we will gain a better understanding of the more general role of this family of proteins in healthy neurons, as well as a better appreciation of the evolutionary significance of this class of protein in neuron function. We will also further our understanding of the consequences of dysfunction of this class of protein as a result of disease or damage. One of the hallmark characteristics of stroke damage and Alzheimer's disease is the dysfunction of the structural microtubule-associated proteins, MAP2 and tau. The consequences of this dysfunction are unknown, because very little is known about how this important class of neuronal cytoskeletal protein functions in healthy neurons. This project will determine the function of one member of this family, PTL-1, in healthy neurons of the nematode, C. elegans, as a way of increasing our understanding of how this class of protein works in normal, healthy adult neurons, which will provide important clues as to the consequences to neurons of the dysfunction of these proteins in disease states.