Microtubule-based transport in lacrimal gland function

泪腺功能中基于微管的运输

• 批准号: 7210566
• 负责人: Sarah F Hamm-Alvarez
• 金额: $ 35.61万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-07-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7210566
• 关键词: 1,2-diacylglycerol; Acinar Cell; Actins; Address; Adenoviruses; Affect; Age; Aging; American; Anatomy; Apical; Atrophic; Autoimmune Process; Autoimmune Responses; Biochemical; Carbachol; Cell membrane; Cells; Clathrin; Communication; Cyan Fluorescent Protein; Cyclophosphamide/Fluorouracil/Prednisone; Cytochalasin D; Cytoskeleton; Data; Development; Diagnosis; Diglycerides; Disease; Dominant-Negative Mutation; Duct (organ) structure; Dynein ATPase; Electron Microscopy; Exhibits; Exocytosis; Experimental Models; Female; Fluorescence; Fluorescence Microscopy; Fluorescence Resonance Energy Transfer; Functional disorder; Gender; Genetic; Gland; Glutathione S-Transferase; Green Fluorescent Proteins; Hormonal; Immune system; Inbred BALB C Mice; Inbred NOD Mice; Individual; Infiltration; Inflammatory Response Pathway; Investigation; Label; Lacrimal gland structure; Lead; Life; Light; Liquid Chromatography; Liquid substance; Lymphocyte; MALDI-TOF Mass Spectrometry; Measures; Mediating; Membrane; Methods; Microfilaments; Microtubules; Molecular; Morbidity - disease rate; Mus; Myosin ATPase; Natural regeneration; Neurons; Non obese; Oryctolagus cuniculus; Pathway interactions; Population; Process; Prolactin; Proline-Rich Domain; Protein Kinase C; Proteins; Proteomics; Recycling; Relative (related person); Resolution; Retrieval; Secretory Vesicles; Sjogren' s Syndrome; Symptoms; Syndrome; Techniques; Testing; Transport Vesicles; VAMP-2; Vesicle; Wiskott-Aldrich Syndrome; Woman; age related; base; basolateral membrane; botulinum toxin type B; diabetic; immunocytochemistry; lacrimal; lacrimal acini; light microscopy; male; mass spectrometer; novel; ocular surface; rat Sycn protein; reconstitution; sex; tandem mass spectrometry; trans-Golgi Network

DESCRIPTION (provided by applicant): A major contribution to ocular morbidity is lacrimal dysfunction, affecting over 10 million Americans, primarily women. At least two to four million cases are autoimmune-mediated and accompanied by additional symptoms, which lead to the diagnosis of Sjogren's syndrome. Development of primary lacrimal deficiency and Sjogren's syndrome are associated with changes in hormonal status and also environmental and genetic factors. Although the precise mechanisms involved in disease development remain unclear, these diseases are associated with impaired release of secretory products into ocular surface fluid by the lacrimal gland. The principal cell of the lacrimal gland and primary contributor of proteins into ocular fluid is the lacrimal acinar cell. Disease-related changes in release of secretory products into ocular fluid imply, at some level, changes in the molecular mechanisms responsible for exocytosis at the apical plasma membrane of lacrimal acini. Our approach to the study of diseases affecting lacrimal gland secretory functions has therefore focused on elucidating the mechanisms responsible for accurate exocytotic release of secretory pro- ducts at the apical plasma membrane of healthy lacrimal acini, and to determine how changes in these mechanisms are related to the establishment and progression of lacrimal disease. Findings during the previous project period have defined the involvement of several key effectors of exocytosis including rab3D, vesicle-associated membrane protein 2, cytoplasmic dynein and actin filaments. Moreover, compensatory retrieval of acinar apical plasma membrane, essential for regeneration of secretory vesicles, utilizes a unique clathrin-mediated endocytotic mechanism involving actin, syndapins and N-WASP. Key questions regarding other fundamental mechanisms of lacrimal acinar exocytotic pathways remain unanswered, and will be addressed in Aims #1 and #2 using reconstituted primary rabbit lacrimal acini as an experimental model. To begin to address the changes in exocytotic pathways associated with initiation and progression of Sjogren's syndrome, Aim #3 probes age- and sex-related changes in organization and composition of the exocytotic pathway in lacrimal glands from control BALB/c mice as well as NOD mice, an experimental model of Sjogren's syndrome. Technically, these aims utilize an array of techniques including light and electron microscopy, biochemical analysis of subcellular membranes and proteomics. The specific aims are: Aim #1. Do lacrimal acinar cells contain distinct secretory vesicle populations? Aim #2. How do actin filaments facilitate exocytosis of secretory vesicles in lacrimal acinar cells? Aim #3. What components of the exocytotic pathway undergo age-related changes in lacrimal glands from BALB/c and NOD mice?

描述（由申请人提供）：对眼部发病率的主要贡献是富有功能障碍，影响了超过1000万美国人，主要是妇女。至少有2至400万例自动免疫介导并伴随着其他症状，这导致了Sjogren综合征的诊断。原发性泪缺乏症和Sjogren综合征的发展与激素状态以及环境和遗传因素的变化有关。尽管疾病发育中涉及的确切机制尚不清楚，但这些疾病与泪腺释放到眼表液中的释放受损有关。泪液中泪腺的主要细胞和蛋白质对眼部液的主要因素是泪腺细胞。疾病相关的分泌产物释放到眼部液中的变化在某种程度上暗示着泪腺上顶质膜上负责胞吐作用的分子机制的变化。因此，我们研究影响泪腺分泌功能的疾病研究的方法是阐明负责准确胞吐释放的机制 在健康泪腺的顶端质膜上的管道，并确定这些机制的变化与富富富疾病的建立和进展如何有关。上一个项目期间的发现定义了胞吞作用的几个关键效应因子，包括RAB3D，囊泡相关的膜蛋白2，细胞质动力蛋白和肌动蛋白丝。此外，对于分泌囊泡的再生必不可少的腺泡根尖质膜的补偿性检索，利用了涉及肌动蛋白，辛达平和N-Wasp的独特网格蛋白介导的内吞作用机制。有关富卓的胞外胞吐途径其他​​基本机制的关键问题尚未得到解答，并将使用重构的主要兔子富富（Aims＃1和＃2）来解决AIMS＃1和＃2，作为一个实验模型。为了开始解决与Sjogren综合征起始和进展相关的胞吐途径的变化，AIM＃3探针在对照Balb/Ciels的核腺体中的组织和性别相关的组织和性别相关的变化以及与NOD小鼠以及NOD小鼠的核囊肿途径的组成，以及Sjogren's Cyndrome cyndrome cyndrome anddrome anddrome cyndrome cyndrome cyndrome cyndrome cynddrome cyndrome cyndrome。从技术上讲，这些目的利用一系列技术，包括光和电子显微镜，亚细胞膜的生化分析和蛋白质组学。具体目的是： 目标＃1。泪腺泡细胞是否包含不同的分泌囊泡群体？ 目标＃2。肌动蛋白丝如何促进泪囊细胞中分泌囊泡的胞吐作用？ 目标＃3。来自BALB/C和NOD小鼠的泪腺的胞吐途径的哪些成分发生与年龄相关的变化？