Patch-Clamp Studies of Mitochondrial Membranes

线粒体膜的膜片钳研究

• 批准号: 8818432
• 负责人: Henry Tedeschi
• 金额: $ 23.08万
• 依托单位: SUNY at Albany
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-04-01 至 1992-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=8818432&HistoricalAwards=false
• 关键词: Patch; Clamp; Studies; Mitochondrial; Membranes

Recent work from this laboratory has provided information on the electrical behavior of mitochondrial membranes by applying patch clamp techniques. The macroscopic currents observed confirmed the presence of voltage sensitive channels in mitochondrial outer membranes previously observed after incorporation of an outer membrane protein (VDAC) into artificial bilayers. In addition, a novel behavior which may correspond to the transient formation of high conductance channels has been described. Completion of these studies and further attempts at obtaining single channel recordings with the outer membrane are proposed. Single channel behavior with the inner membrane has been observed in this laboratory using voltage clamp techniques. Multiple conductance levels are observed and a primary goal of this proposal is to characterize these levels in terms of their channel behavior, e.g. ion selectivity. The general macroscopic ionic currents associated with the initiation of metabolism include a) a decrease in resistance which is reversed by a metabolic inhibitor, b) a change in outward current, and c) a change in the magnitude of current fluctuations. Proposed work includes investigation of the underlying events responsible for these observations. In addition, the effect of imposed membrane potentials on ATP synthesis will be studied. Mitochondria are organelles within cells where biochemical reactions convert energy into a useful form that supports many other cell functions. The biochemical reactions going on in mitochondria have been the object of much research over many years. It is now generally believed that an important aspect of the regulation of mitochondrial function involves the distribution of ions across the mitochondrial membranes. "Patch clamp" techniques of electrophysiology that permit measurement of the permeability pathways for ions across the mitochondrial membrane have become available only relatively recently. The proposed research represents one of the first systematic studies of the permeability properties of intact mitochondrial membranes taking advantage of this technique. The results of this research will make an important contribution to basic knowledge of the function of these organelles.

该实验室最近的工作提供了以下信息 线粒体膜的电行为， 膜片钳技术。 观察到的宏观电流 证实存在电压敏感通道， 线粒体外膜以前观察到后， 将外膜蛋白（VDAC）掺入人工 双层。 此外，一种新的行为，可能对应于 高电导通道的瞬时形成 介绍了 完成这些研究并进一步尝试 用外膜获得单通道记录是 提出了 单通道行为与内膜有 在这个实验室里用电压钳技术观察到的。 观察到多个电导水平， 这一建议是根据这些水平的特点， 通道行为，例如离子选择性。 一般宏观 与代谢起始相关的离子电流 包括：a）电阻减小，其被 代谢抑制剂，B）外向电流的变化，和c）a 电流波动幅度的变化。 拟议工作 包括调查导致 这些观察。 此外，施加膜的效果 将研究ATP合成的潜力。 线粒体是细胞内的细胞器， 反应将能量转化为有用的形式， 其他细胞功能。 体内发生的生化反应 线粒体一直是许多研究的对象， 年 现在人们普遍认为， 线粒体功能的调节涉及 离子在线粒体膜上的分布。 "补丁 电生理学的"钳"技术，允许测量 离子穿过线粒体的渗透性途径 膜只是相对最近才变得可用。 的 拟议的研究代表了第一个系统的研究之一， 完整的线粒体膜的渗透性 利用这种技术。 这项研究成果 将对人类的基本知识做出重要贡献， 这些细胞器的功能。