Differentiation Induced Changes in Centrosomes and Microtubule Organization

分化诱导中心体和微管组织的变化

• 批准号: 9121588
• 负责人: Terry H Lechler
• 金额: $ 29.78万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9121588
• 关键词: Address; Alleles; Architecture; Binding Proteins; Biology; Cell Adhesion; Cells; Centrosome; Complex; Cultured Cells; Cytoplasm; Data; Defect; Disease; Epidermis; Epithelial; Epithelium; Generations; Genetic Models; Goals; Grant; Health; Imagery; Intestines; Lead; Maintenance; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Microtubule Proteins; Microtubules; Minus End of the Microtubule; Molecular; Mus; Muscle; Mutant Strains Mice; Neurodegenerative Disorders; Neurons; Pathology; Pathway interactions; Physiological; Physiology; Polymers; Population; Process; Proteins; Protocols documentation; Radial; Reagent; Recruitment Activity; Role; Simple Epithelium; Site; Skin; Structure; Technology; Tissues; Transgenic Mice; Tubulin; Vesicle; Work; base; cell type; ciliopathy; gastrointestinal epithelium; mutant; ninein; novel; protocol development; scaffold; tool; trafficking

DESCRIPTION (provided by applicant): Differentiation induces a change in microtubule organization in cell types as diverse as epithelia, muscles and neurons. This requires loss of the centrosome's microtubule organizing activity and formation of non- centrosomal microtubule arrays. We have a very limited understanding of how this reorganization occurs and the functions of the resulting microtubules. In part, this is due to lack of tools to observe and specifically perturb microtubule organization in tissues. The long-term goal of our work is to understand how diverse microtubule organizations are generated in differentiated cells and to expose their functions. The objectives of this proposal are to determine how loss of centrosomal organizing activity and microtubule minus end anchoring proteins collaborate to induce specific microtubule arrays. Based on our preliminary data, we hypothesize that differentiation causes changes in both nucleation and anchoring of microtubules at centrosomes. In combination with cytoplasmic minus end anchoring proteins, this allows for specific cytoskeletal structures to form in differentiated cells. We have developed tools/technologies that will allow us to fully characterize the changes in centrosome nucleation activity, ultrastructure and protein composition that occur upon differentiation in the epidermis. In addition, we will elucidate the underlying molecular mechanisms that lead to changes in centrosome composition and activity. Finally, we have developed novel mouse lines that will be used to examine the roles of noncentrosomal microtubule arrays in both the skin and the intestine through specific disruption of microtubule minus end anchoring proteins. These studies will elucidate the mechanisms regulating centrosome activity and microtubule reorganization in differentiated cells and determine the physiologic functions of these arrays in two different tissues.

描述（由申请人提供）：分化诱导细胞类型（如上皮细胞、肌肉和神经元）中微管组织的变化。这需要中心体微管组织活性的丧失和非中心体微管阵列的形成。我们对这种重组如何发生以及所产生的微管的功能的理解非常有限。在某种程度上，这是由于缺乏工具来观察和专门扰乱组织中的微管组织。我们工作的长期目标是了解不同的微管组织是如何在分化的细胞中产生的，并揭示它们的功能。这个建议的目的是确定如何损失的中心体组织活动和微管减去端锚定蛋白合作，以诱导特定的微管阵列。根据我们的初步数据，我们假设分化会导致中心体微管的成核和锚定发生变化。与细胞质负端锚定蛋白结合，这允许在分化的细胞中形成特定的细胞骨架结构。我们已经开发了工具/技术，使我们能够充分表征表皮分化后发生的中心体成核活性，超微结构和蛋白质组成的变化。此外，我们将阐明导致中心体组成和活性变化的潜在分子机制。最后，我们已经开发了新的小鼠品系，将用于检查的作用，非中心体微管阵列在皮肤和肠道通过特定的破坏微管减去端锚定蛋白。这些研究将阐明在分化细胞中调节中心体活性和微管重组的机制，并确定这些阵列在两种不同组织中的生理功能。