High accuracy function analysis of mitochondria with organelle adsorption method

细胞器吸附法高精度线粒体功能分析

• 批准号: 17570129
• 负责人: OHTA Yoshihiro
• 金额: $ 2.24万
• 依托单位: Tokyo University of Agriculture and Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17570129/
• 关键词: mitochondria; biophysics; bioimaging; reactive oxygen species; uncoupling protein 2; cell death; UCP2; 膜電位; 酸化ストレス; 神経細胞

Purpose:Developments of the method for measuring organelles' functions are not enough, although their functions are very important. Organelle adsorption method is a newly developed method which enables us to measure the activity of organelles adsorbed on the base. The characteristics are the measurements of individual organelles and the quick separation of the metabolites from the organelle. In this study, we measured mitochondrial behaviors with organelle adsorption method. Results and Discussions:We measured the effects of 1) the uncoupling protein 2 (UCP2), 2) cyclophilin D,3) ganglioside GD3, 4) Ca2+, 5) respiration substrates on mitochondrial behaviors, and published 4 original papers in international journal and one review in domestic journal. The results are 1) UCP2 and cyclophilin D protect mitochondria from oxidative stress-induced damage, 2) GD3 promotes the generation of reactive oxygen species from mitochondria and induces the gradual depolarization, 3) mitochondrial membrane potential are stabilized in the cell by the presence of ADP, ATP and Mg2+, 4) Ca2+ induced the swelling of depolarized mitochondria not but polarized mitochondria. These results provide us the important information on mitochondrial behaviors.

目的：细胞器的功能虽然十分重要，但其功能测定方法的发展还不够。细胞器吸附法是近年来发展起来的一种测定细胞器活性的新方法。其特点是单个细胞器的测量和代谢产物从细胞器的快速分离。本研究用细胞器吸附法测定了线粒体的行为。结果与讨论：我们检测了解偶联蛋白2（UCP 2）、亲环素D（cyclophilin D）、神经节苷脂GD 3（ganglioside GD 3）、钙离子（Ca 2+）、呼吸底物对线粒体行为的影响，在国际期刊上发表了4篇原创论文，在国内期刊上发表了1篇综述。结果表明：1）UCP 2和亲环素D对氧化应激引起的线粒体损伤具有保护作用; 2）GD 3促进线粒体产生活性氧，并使线粒体逐渐去极化; 3）ADP、ATP和Mg ~（2+）稳定线粒体膜电位;这些结果为我们研究线粒体的行为提供了重要的信息。

项目成果

Enhancementof nitric oxide production by association of nitric oxidesynthase with N-methyl-d-aspartate receptors viapostsynaptic density 95 in genetically engineeredChinese hamster ovary cells : real-time fluorescenceimaging using nitric oxide sensitive d

在基因工程中国仓鼠卵巢细胞中，通过突触后密度 95 将一氧化氮合酶与 N-甲基-d-天冬氨酸受体结合，增强一氧化氮的产生：使用一氧化氮敏感 d 进行实时荧光成像

• 发表时间: 2006
• 期刊: Journal of Neurochemistry 96
• 作者: Ishii;H.;Shibuya;K.;Ohta;Y.;Mukai;H.;Uchino;S.;Takata;N.;Rose;J.A.;Kawato;S.
• 通讯作者: S.

Stability of membrane potential in heart mitochondria: single mitochondrion imaging.

• DOI: 10.1016/j.bbrc.2006.03.233
• 发表时间: 2006-06
• 期刊: Biochemical and biophysical research communications
• 影响因子: 3.1
• 作者: Yukiko Uechi;H. Yoshioka;D. Morikawa;Y. Ohta

Effects of disialoganglioside GD3 on the mitochondrial membrane potential

• DOI: 10.1016/j.febslet.2005.04.054
• 发表时间: 2005-06-06
• 期刊: FEBS LETTERS
• 影响因子: 3.5
• 作者: Higuchi, Y;Miura, T;Ohta, Y
• 通讯作者: Ohta, Y

Enhancement of nitric oxide production by association of nitric oxide synthase with N-methyl-d-aspartate receptors via postsynaptic density 95 in genetically engineered Chinese hamster ovary cells : real-time fluorescence imaging using nitric oxide sensit

在基因工程中国仓鼠卵巢细胞中，一氧化氮合酶通过突触后密度 95 与 N-甲基-d-天冬氨酸受体结合，增强一氧化氮的产生：使用一氧化氮敏感性的实时荧光成像

• 发表时间: 2006
• 期刊: Journal of Neurochemistry 96
• 作者: Ishii;H.;Shibuya;K.;Ohta;Y.;Mukai;H.;Uchino;S.;Takata;N.;Rose;J.A.;Kawato;S.
• 通讯作者: S.

Developments of new technique for organellemeasurement and application for mitochondria

线粒体细胞器测量新技术及其应用进展

• 发表时间: 2005
• 期刊: Bioimaging 14
• 作者: Higuchi;Y.;Miura;T.;Kajimoto;T.;Ohta;Y.;Ohta Y
• 通讯作者: Ohta Y