ULTRASTRUCTURE OF APOPTOTIC MITOCHONDRIA

凋亡线粒体的超微结构

• 批准号: 7601022
• 负责人: DONALD DAVID NEWMEYER
• 金额: $ 1.09万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7601022
• 关键词: Apoptosis; Apoptotic; Biological Preservation; Buffers; Carbohydrates; Collaborations; Computer Retrieval of Information on Scientific Projects Database; Cytoplasm; Freezing; Funding; Grant; In Vitro; Institution; Liver Mitochondria; Mannitol; Membrane; Methods; Mitochondria; Mus; Organelles; Outer Mitochondrial Membrane; Pathway interactions; Preparation; Process; Protein Family; Proteins; Research; Research Personnel; Resources; Rupture; Source; Standards of Weights and Measures; Sucrose; Time; Trehalose; United States National Institutes of Health; Work; controlled release; research study

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We are investigating a method for the long-term storage of mitochondria in trehalose-containing buffer. In apoptosis, Bcl-2-family proteins regulate the barrier function of the mitochondrial outer membrane (MOM), controlling the release of proapoptotic proteins from the intermembrane space (IMS) into the cytoplasm. This process can be studied in vitro with freshly isolated mouse liver mitochondria. Unfortunately, mitochondria frozen/thawed in standard sucrose-mannitol buffers become leaky and useless for apoptosis research. Much of what we know about the mitochondrial apoptotic pathway has been determined from experiments using isolated mouse liver mitochondria. The outer membranes of these isolated organelles tend to deteriorate over time and rupture immediately upon freezethaw. Thus, the functional analysis of mouse liver mitochondria (and presumably mammalian mitochondria from other sources) currently requires freshly prepared organelles. To reduce the need for costly and timeconsuming preparations, we sought to identify a storage medium that preserves mitochondrial function, particularly of the outer membrane. To that end, we analyzed the integrity of MOMs after rapid freezethaw in buffers containing various carbohydrates. Part of this analysis was to determine the structural preservation of isolated mitochondria. This work was done at the NCMIR in collaboration with Dr. Perkins.

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