Giant Mitochondria in Aging

衰老过程中的巨型线粒体

基本信息

批准号：
7432916
负责人：
EDGAR A ARRIAGA
金额：
$ 2.32万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-04-15 至 2012-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7432916
关键词：
3-methyladenine Affect Age Aging Alcoholism Amino Acids Appearance Autophagocytosis Biological Biological Aging Capillary Electrophoresis Cell model Cells Cellular biology Centrifugation Characteristics Class Collaborations Confocal Microscopy Cultured Cells Defect Disease Electrons Electrophoresis Fiber Fluorescence Microscopy Funding Gene Mutation Genomics Goals Health Human Hyperoxia Individual Knowledge Label Link Literature Membrane Potentials Methods Mitochondria Mitochondrial DNA Mitotic Modeling Molecular Muscle Muscle Fibers Muscle function Myoblasts Organelles Performance Preparation Principal Investigator Procedures Production Property Proteins Proteome Proteomics Public Health Rattus Relative (related person)Reporting Research Resources Role Sampling Scientist Skeletal Muscle Stable Isotope Labeling Techniques Technology Time Tissues Toxic effect Work age group age related base in vivo new technology novel protein expression size theories tool

项目摘要

There is significant evidence linking mitochondria to biological aging, with a recent theory proposing that giant mitochondria are involved in the aging of post-mitotic tissues. In these tissues, giant mitochondria appear to evade autophagocytosis, accumulate, and replace normal mitochondria. Techniques to characterize giant mitochondria in the presence of, or after isolation from, normal mitochondria are greatly needed. The short- term goal of this proposal is to characterize and isolate giant mitochondria and establish whether or not these mitochondria affect muscle function. The long-term goal is to use and make available novel technological approaches for uncovering the changes that, at the subcellular and molecular level, define biological human aging. The proposed work utilizes mitochondria from a myoblast model and muscle from Fischer 344 rats. We have four aims: (1) Characterize giant mitochondria in vivo in cell models based on their cell biology. (2) Purify giant mitochondria isolated from these in vivo cell models and then characterize these giant mitochondria biochemically. (3) Compare the proteomes of giant and normal mitochondria isolated from these models. (4) Characterize giant mitochondria in aging skeletal muscle. The first aim uses confocal microscopy, or this technique combined with single cell capillary electrophoresis, to investigate size, DMAcontent and synthesis, DNA mutations, carbonyl accumulation, ROS production, membrane potential, mitochondrial fusion, and autophagocytosis. The second aim describes the methods to isolate subpopulations of giant and normal mitochondria using free-flow electrophoresis and then characterize these isolated mitochondria using methods suitable for isolated organelles. The third aim utilizes stable isotope labeling in cell cultures to compare the proteomes of giant and normal mitochondria. The last aim investigates age-related changes in muscle containing giant mitochondria. Significance to Health. Giant mitochondria remain poorly characterized, their proteomic and genomic characteristics have not been defined, and their relationship to muscle function has not been investigated. The characterization of this class of mitochondria is highly relevant to the public health not only because of their possible role in aging tissues, but also because of their appearance as a consequence of alcoholism, human toxicity, and multiple diseases.

有重要证据表明线粒体与生物衰老有关，最近的一项理论提出，线粒体与生物衰老有关。线粒体参与有丝分裂后组织的衰老。在这些组织中，巨大的线粒体似乎逃避自噬作用，积累并取代正常线粒体。表征巨人的技术非常需要存在正常线粒体或从正常线粒体分离后的线粒体。短- 该提案的长期目标是表征和分离巨型线粒体，并确定这些是否线粒体影响肌肉功能。长期目标是使用并提供新技术揭示在亚细胞和分子水平上定义生物人类的变化的方法老化。拟议的工作利用来自成肌细胞模型的线粒体和来自 Fischer 344 大鼠的肌肉。我们有四个目标：（1）根据细胞生物学特征在细胞模型中表征体内巨型线粒体。 (2)净化从这些体内细胞模型中分离出巨型线粒体，然后表征这些巨型线粒体生物化学上。 (3) 比较从这些模型中分离的巨型线粒体和正常线粒体的蛋白质组。 (4) 表征衰老骨骼肌中巨大的线粒体。第一个目标是使用共焦显微镜，或者这个技术与单细胞毛细管电泳相结合，研究尺寸、DMA 含量和合成， DNA 突变、羰基积累、ROS 产生、膜电位、线粒体融合和自噬作用。第二个目标描述了分离巨型和正常亚群的方法使用自由流动电泳分析线粒体，然后使用以下方法表征这些分离的线粒体适用于分离细胞器的方法。第三个目标利用细胞培养物中的稳定同位素标记比较巨型线粒体和正常线粒体的蛋白质组。最后一个目标是调查与年龄相关的变化含有巨大线粒体的肌肉。对健康的意义。巨型线粒体的特征仍知之甚少，其蛋白质组学和基因组学特征尚未定义，它们与肌肉功能的关系尚未研究。这类线粒体的表征与公众健康高度相关，不仅因为它们可能在组织老化中发挥作用，而且还因为它们的出现是酗酒的结果，人类毒性和多种疾病。