Quantitating Mitophagy in Skeletal Muscle Models

骨骼肌模型中线粒体自噬的定量

• 批准号: 8822184
• 负责人: EDGAR A ARRIAGA
• 金额: $ 40.19万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-15 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8822184
• 关键词: Address; Aftercare; Aging; Amino Acids; Animal Model; Autophagocytosis; Autophagosome; Biopsy; Capillary Electrophoresis; Cell Culture Techniques; Cell model; Cell physiology; Cells; Chemicals; Clinic; Clinical Research; Degradation Pathway; Detection; Diabetes Mellitus; Disease; Elderly; Excision; Fluorescence; Functional disorder; Goals; Hormones; Human; Hyperthyroidism; Individual; Label; Lasers; Malignant Neoplasms; Measurement; Measures; Mediating; Mitochondria; Modeling; Molecular Chaperones; Monitor; Muscle; Muscle Fibers; Myoblasts; Organelles; Outcome; Oxidative Stress; Parkinson Disease; Photosensitization; Play; Process; Proteome; Proteomics; Rattus; Relative (related person); Ribosomes; Role; Skeletal Muscle; Specificity; Speed; Stable Isotope Labeling; Starvation; Subcellular structure; System; Techniques; Technology; Testing; Thyroid Hormones; Time; Tissue Model; Triiodothyronine; Ubiquitination; Western Blotting; Work; animal tissue; base; design; human tissue; inhibitor/antagonist; muscle form; new technology; oxidative damage; peroxisome; protein aggregate; protein degradation; trend

DESCRIPTION (provided by applicant): Mitophagy is a specialized form of macroautophagy responsible for removal of non-functional mitochondria from the cell. Given the prominent role that mitochondria play in oxidative stress, it has been suggested that mitophagy is critical to protein turnover in skeletal muscle. It is currently impossible to determine how each form of autophagy, including mitophagy, contributes to protein turnover. The immediate goal of this application is to develop bioanalytical approaches to determine mitophagy fluxes in cell cultures and skeletal muscle. The main parameter to measure is mitophagy flux (the rate at which mitophagy occurs) using novel technologies based on individual organelle measurements by capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) and proteomic profiles. The long-term goal is to apply these bioanalytical approaches to characterize the role of mitophagy in human conditions such as hyperthyroidism and aging that are characterized by alterations in protein turnover. The specific aims of this application are: 1. Define new mitophagy flux analyses based on individual organelle measurements by CE-LIF. 2. Identify organelle-specific proteome changes in abundance, carbonylation and ubiquitination that are associated with changes in mitophagy flux. 3. Determine changes in mitophagy flux in skeletal muscle of hyperthyroid rat, aging rat and aging human models. This work approaches mitophagy from a unique angle, aiming to integrate individual organelle measurements and proteome profiles to define altered mitophagy fluxes with high specificity, even when in the presence of other forms of autophagy. We will the transfer the new technologies to a clinical research setting to investigate mitophagy in human skeletal muscle. Long-term impact of the findings of this work would be the basis for investigating mechanisms of mitophagy and its role in disease and aging.

描述（由申请人提供）：线粒体是一种专门的大型噬菌体形式，负责从细胞中去除非功能性线粒体。鉴于线粒体在氧化应激中发挥作用的重要作用，因此有人提出线粒体对骨骼肌的蛋白质更新至关重要。目前无法确定每种形式的自噬如何，包括线粒体，都会导致蛋白质更新。该应用的直接目标是开发生物分析方法，以确定细胞培养和骨骼肌中的线粒体通量。要测量的主要参数是使用新技术基于单个细胞器测量的新技术，该技术是通过激光诱导的荧光检测（CE-LIF）和蛋白质组曲线的。长期目标是应用这些生物分析方法来表征 在人类条件下的线粒体，例如甲状腺功能亢进和衰老，其特征是蛋白质更新的改变。 该应用的具体目的是：1。根据CE-LIF的单个细胞器测量来定义新的线粒体通量分析。 2。确定与线粒体通量变化有关的丰度，羰基化和泛素化的蛋白质组变化。 3。确定甲状腺功能亢进症，衰老大鼠和衰老人类模型的骨骼肌中线粒体通量的变化。这项工作从独特的角度接近线粒体，旨在整合单个细胞器的测量和蛋白质组轮廓，以定义具有高特异性的线粒体通量，即使在存在其他形式的自噬的情况下。我们将把新技术转移到临床研究环境中，以研究人类骨骼肌的线粒体。这项工作结果的长期影响将是研究线粒体机制及其在疾病和衰老中的作用的基础。